Methods of quantifying oligosaccharide preparations

Abstract

The present disclosure relates to selective analytical methods for the detection and / or quantification of an oligosaccharide preparation in a nutritional composition such as an animal feed. Also disclosed are methods of manufacturing a nutritional composition comprising an oligosaccharide preparation, the presence or concentration of which can be selectively detected or determined.

Description

METHODS OF QUANTIFYING OLIGOSACCHARIDE PREPARATIONS CROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 757,231 filed on November 8, 2018. BACKGROUND

[0002] Oligosaccharide preparations, which can generally include monosaccharides, oligosaccharides, polysaccharides, functionalized oligosaccharides, or their combinations, are used as additives in nutritional compositions such as animal feed. The addition of oligosaccharide preparations can improve the health and performance of the animal. However, it is challenging to detect or quantify an oligosaccharide preparation additive in a nutritional composition, because nutritional compositions usually contain other carbohydrate sources that can have structural similarities with the oligosaccharide preparations. As a result, a need exists for methods of selectively detecting or quantifying the oligosaccharide preparations in a nutritional composition. SUMMARY

[0003] Disclosed herein are simple, selective, and sensitive analytical methods for the detection and / or quantification of an oligosaccharide preparation or a composition comprising the oligosaccharide preparation in a nutritional composition. Also disclosed are methods of manufacturing a nutritional composition comprising an oligosaccharide preparation, the presence or concentration of which can be selectively and sensitively detected or determined.

[0004] In one aspect, described herein is a method of correlating a synthetic oligosaccharide preparation in a nutritional composition, wherein the nutritional composition comprises the synthetic oligosaccharide preparation and a naturally occurring oligosaccharide composition, the method comprising: (a) providing a sample of the nutritional composition, (b) detecting a signal of at least a portion of oligosaccharides in the sample of the nutritional composition, and (c) correlating a concentration of the synthetic oligosaccharide preparation in the nutritional composition, wherein the signal is (i) indicative of one or more anhydro-subunit containing oligosaccharides or (ii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>- (1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In one aspect, described herein is a method of performing quality control of a nutritional composition comprising: (a) providing a batch of a nutritional composition, wherein the nutritional composition comprises a synthetic oligosaccharide preparation and a naturally occurring oligosaccharide composition, (b) obtaining a sample of the nutritional composition from the batch, (c) detecting a signal of at least a portion of oligosaccharides in the sample of the nutritional composition through analytical instrumentation, and (d) accepting or rejecting the batch of the nutritional composition, wherein the signal is (i) indicative of one or more anhydro-subunit containing oligosaccharides or (ii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In some embodiments, the signal is indicative of one or more anhydro-subunit containing oligosaccharides. In some embodiments, the one or more anhydro-subunit containing oligosaccharides have a degree of polymerization of 2 (DP2). In one aspect, described herein is a method of performing quality control of a nutritional composition comprising: (a) providing a sample of a nutritional composition, wherein the nutritional composition comprises a naturally occurring oligosaccharide composition, and (b) detecting a signal of at least a portion of oligosaccharides in the sample of the nutritional composition through analytical instrumentation, wherein the signal is indicative of one or more anhydro-subunit containing oligosaccharides having a degree of polymerization of 2 (DP2). In some embodiments, the nutritional composition comprises a synthetic oligosaccharide preparation. In some embodiments, the method comprises correlating a concentration of the synthetic oligosaccharide preparation in the nutritional composition. In some embodiments, the signal is detected by high-performance liquid chromatography (HPLC), gas chromatography (GC), mass spectrometry (MS), nuclear magnetic resonance (NMR) spectroscopy, size exclusion chromatography (SEC), field flow fractionation (FFF), asymmetric flow field flow fractionation (A4F), weight determination of fractions by preparative chromatography, or any combination thereof. In some embodiments, the nutritional composition comprises a base nutritional composition. In some embodiments, the base nutritional composition comprises the naturally occurring oligosaccharide composition. In some embodiments, the base nutritional composition lacks a detectable level of anhydro-subunit containing oligosaccharides. In some embodiments, the base nutritional composition is essentially free of anhydro-subunit containing oligosaccharides. In some embodiments, the one or more anhydro-subunit containing oligosaccharides originate from the synthetic oligosaccharide preparation. In some embodiments, the signal is attributed to anhydro-subunit containing oligosaccharides having a degree of polymerization of 1 (DP1). In some embodiments, the signal is attributed to levoglucosan, 1,6-anhydro-β-D-glucofuranose, or a combination thereof. In some embodiments, the signal is attributed to anhydro-subunit containing oligosaccharides having a degree of polymerization of 3 (DP3). In some embodiments, the signal is attributed to DP1, DP2, or DP3 anhydro-subunit containing oligosaccharides, or a combination thereof. In some embodiments, the signal is attributed to one or more DP2 anhydro-subunit containing oligosaccharides. In some embodiments, the signal is attributed to anhydro-cellobiose. In some embodiments, the detecting comprises a weight determination of one or more degree of polymerization (DP) fractions of oligosaccharides. In some embodiments, the detecting comprises a weight determination of at least a portion of anhydro-subunit containing oligosaccharides from the sample. In some embodiments, the at least a portion of anhydro-subunit containing oligosaccharides have a degree of polymerization of 1, 2, or 3. In some embodiments, the one or more DP fractions of oligosaccharides or the at least a portion of anhydro-subunit containing oligosaccharides are isolated by preparative chromatography. In some embodiments, the signal is detected, at least in part, by matrix-assisted laser desorption / ionization-mass spectrometry (MALDI-MS). In some embodiments, the signal is detected, at least in part, by liquid chromatography—mass spectrometry (LC-MS) / MS. In some embodiments, the signal is detected, at least in part, by GC-flame ionization detector (GC-FID) or GC-MS. In some embodiments, the signal is detected, at least in part, by NMR spectroscopy. In some embodiments, the signal is associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages of oligosaccharides. In some embodiments, the detecting comprises obtaining an NMR spectroscopy. In some embodiments, the signal is associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages. In some embodiments, the signal is associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages. In some embodiments, the signal is associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages. In some embodiments, the signal is associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages. In some embodiments, the signal is associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages. In some embodiments, the signal is associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages. In some embodiments, the signal is associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages. In some embodiments, the detecting comprises determining the presence or absence of the signal. In some embodiments, the detecting comprises determining the presence or absence of DP1 or DP2 anhydro-subunit containing oligosaccharides, or both. In some embodiments, the detecting comprises determining or correlating a level of the signal. In some embodiments, the detecting comprises correlating a level of DP1 anhydro-subunit containing oligosaccharides in the nutritional composition. In some embodiments, the detecting comprises correlating a level of DP2 anhydro-subunit containing oligosaccharides in the nutritional composition.

[0005] In one aspect, described herein is a method of performing quality control of a nutritional composition comprising a synthetic oligosaccharide preparation and a naturally occurring oligosaccharide composition, the method comprising: (a) providing a first sample of the nutritional composition, (b) providing a second sample of the nutritional composition, (c) detecting a first signal of at least a portion of oligosaccharides in the first sample, (d) detecting a second signal of at least a portion of oligosaccharides in the second sample, and (e) comparing the first signal and the second signal, wherein the first signal and the second signal are independently (i) indicative of one or more anhydro-subunit containing oligosaccharides or (ii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In some embodiments, the method comprises correlating a concentration of the synthetic oligosaccharide preparation in the nutritional composition of the first sample, a concentration of the synthetic oligosaccharide preparation in the nutritional composition of the second sample, or both. In some embodiments, the first signal and the second signal are each independently indicative of one or more anhydro- subunit containing oligosaccharides. In some embodiments, the first signal and the second signal are attributed to the same species of an anhydro-subunit containing oligosaccharide. In some embodiments, the first signal and the second signal are attributed to different species of anhydro- subunit containing oligosaccharides In some embodiments, the first sample and the second sample are taken from different batches of the nutritional composition. In some embodiments, the first signal and the second signal are each independently detected by high-performance liquid chromatography (HPLC), gas chromatography (GC), mass spectrometry (MS), nuclear magnetic resonance (NMR) spectroscopy, size exclusion chromatography (SEC), field flow fractionation (FFF), asymmetric flow field flow fractionation (A4F), weight determination of fractions by preparative chromatography, or any combination thereof. In some embodiments, the nutritional composition comprises a base nutritional composition. In some embodiments, the base nutritional composition comprises the naturally occurring oligosaccharide composition. In some embodiments, the base nutritional composition lacks a detectable level of anhydro-subunit containing oligosaccharides. In some embodiments, the base nutritional composition is essentially free of anhydro-subunit containing oligosaccharides. In some embodiments, the one or more anhydro-subunit containing oligosaccharides originate from the synthetic oligosaccharide preparation. In some embodiments, the first signal, the second signal, or both are independently attributed to anhydro-subunit containing oligosaccharides having a degree of polymerization of 1 (DP1). In some embodiments, the first signal, the second signal, or both are independently attributed to levoglucosan, 1,6-anhydro-β-D-glucofuranose, or a combination thereof. In some embodiments, the first signal, the second signal, or both are independently attributed to anhydro- subunit containing oligosaccharides having a degree of polymerization of 2 (DP2). In some embodiments, the signal is attributed to anhydro-cellobiose. In some embodiments, the first signal, the second signal, or both are independently attributed to anhydro-subunit containing oligosaccharides having a degree of polymerization of 3 (DP3). In some embodiments, the first signal, the second signal, or both are independently attributed to DP1, DP2, or DP3 anhydro- subunit containing oligosaccharides, or a combination thereof. In some embodiments, the detecting comprises a weight determination of one or more degree of polymerization (DP) fractions from the sample or the second sample. In some embodiments, the detecting comprises a weight determination of at least a portion of anhydro-subunit containing oligosaccharides from the first sample or the second sample. In some embodiments, the at least a portion of anhydro- subunit containing oligosaccharides have a degree of polymerization of 1, 2, or 3. In some embodiments, the one or more DP fractions of oligosaccharides or the at least a portion of anhydro-subunit containing oligosaccharides are isolated by preparative chromatography. In some embodiments, the first signal, the second signal, or both are independently detected, at least in part, by matrix-assisted laser desorption / ionization-mass spectrometry (MALDI-MS). In some embodiments, the first signal, the second signal, or both are independently detected, at least in part, by liquid chromatography—mass spectrometry (LC-MS) / MS. In some embodiments, the first signal, the second signal, or both are independently detected, at least in part, by GC-flame ionization detector (GC-FID) or GC-MS. In some embodiments, the first signal, the second signal, or both are independently detected, at least in part, by NMR spectroscopy. In some embodiments, the first signal, the second signal, or both are independently associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages of oligosaccharides. In some embodiments, the detecting comprises obtaining an NMR spectroscopy. In some embodiments, the first signal, the second signal, or both are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages. In some embodiments, the first signal, the second signal, or both are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages. In some embodiments, the first signal, the second signal, or both are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages. In some embodiments, the first signal, the second signal, or both are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages. In some embodiments, the first signal, the second signal, or both are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages. In some embodiments, the first signal, the second signal, or both are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,4) glycosidic linkages. In some embodiments, the first signal, the second signal, or both are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages. In some embodiments, the first signal is indicative of one or more anhydro-subunit containing oligosaccharides and the second signal is associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In some embodiments, the detecting comprises determining the presence or absence of the first signal and the second signal. In some embodiments, the detecting comprises determining the presence or absence of DP1 or DP2 anhydro-subunit containing oligosaccharides, or both. In some embodiments, the detecting comprises determining or correlating a level of the first signal and the second signal. In some embodiments, the detecting comprises correlating a level of DP1 anhydro-subunit containing oligosaccharides in the nutritional composition. In some embodiments, the detecting comprises correlating a level of DP2 anhydro-subunit containing oligosaccharides in the nutritional composition.

[0006] In some embodiments, a method described herein comprises accepting or rejecting a batch of the nutritional composition. In some embodiments, the method comprises adjusting a level of the synthetic oligosaccharide preparation in the nutritional composition after the detecting. In some embodiments, the base nutritional composition comprises a plurality of oligosaccharides. In some embodiments, the base nutritional composition comprises starch or plant fibers. In some embodiments, a level of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkage, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkage, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkage in the base nutritional composition is at least 10% lower than a level of the same glycosidic linkage in the synthetic oligosaccharide preparation. In some embodiments, the method comprises a derivatization step prior to the detecting. In some embodiments, the method comprises extracting oligosaccharides from the sample of the nutritional composition. In some embodiments, the method comprises filtering or clarifying the extracted oligosaccharides. In some embodiments, the method comprises concentrating the extracted oligosaccharides. In some embodiments, the concentrating comprises freeze-drying. In some embodiments, the concentrating comprises nano-filtration. In some embodiments, the method comprises introducing an internal standard into the extracted or concentrated oligosaccharides. In some embodiments, the method comprises reducing the extracted or concentrated oligosaccharides. In some embodiments, the method comprises digesting the extracted or concentrated oligosaccharides with one or more hydrolytic enzymes. In some embodiments, the one or more hydrolytic enzymes comprise carbohydratase, protease, lipase, or any combination thereof. In some embodiments, the one or more hydrolytic enzymes comprise <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-amylase, amyloglycosidase, invertase, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-galactosidase, or any combination thereof. In some embodiments, the one or more hydrolytic enzymes cleave one or more naturally occurring glycosidic bonds. In some embodiments, the method comprises isolating the oligosaccharides that are not digested. In some embodiments, the method comprises separating the extracted, concentrated, digested, or reduced oligosaccharides. In some embodiments, the oligosaccharides are separated chromatographically. In some embodiments, the method comprises isolating the separated oligosaccharides. In some embodiments, the oligosaccharides are separated or isolated by their degrees of polymerization. In some embodiments, the method comprises isolating or separating the oligosaccharides with a degree of polymerization of 1, 2, 3, 4, or 5. In some embodiments, the DP1 oligosaccharides are isolated or separated. In some embodiments, the DP2 oligosaccharides are isolated or separated. In some embodiments, the DP3 oligosaccharides are isolated or separated. In some embodiments, the method comprises isolating at least a portion of the DP1 anhydro-subunit containing oligosaccharides from the sample. In some embodiments, the method comprises isolating at least a portion of the DP2 anhydro-subunit containing oligosaccharides from the sample. In some embodiments, the method comprises isolating at least a portion of the DP3 anhydro-subunit containing oligosaccharides from the sample. In some embodiments, the isolated or separated oligosaccharides are quantified. In some embodiments, a majority of the quantified oligosaccharides originate from the synthetic oligosaccharide preparation. In some embodiments, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, or more than 99% by weight of the quantified oligosaccharide originate from the synthetic oligosaccharide preparation.

[0007] In one aspect, described herein is a method of correlating a synthetic oligosaccharide preparation in a nutritional composition, wherein the nutritional composition comprises (i) a synthetic oligosaccharide preparation that comprises anhydro-subunit containing oligosaccharides and (ii) a naturally occurring oligosaccharide composition, the method comprising: (a) providing a sample of the nutritional composition, (b) isolating one or more anhydro-subunit containing oligosaccharides from the sample, (c) detecting a signal that is indicative of the one or more anhydro-subunit containing oligosaccharides, wherein the detecting comprises (i) a weight determination of at least a portion of anhydro-subunit containing oligosaccharides from the sample or (ii) analyzing at least a portion of anhydro-subunit containing oligosaccharides from the sample by matrix-assisted laser desorption / ionization-mass spectrometry (MALDI-MS), liquid chromatography—mass spectrometry (LC-MS) / MS, or gas chromatography(GC)-MS, and (d) correlating a concentration of the synthetic oligosaccharide preparation in the nutritional composition. In some embodiments, the detecting comprises a weight determination of at least a portion of the anhydro-subunit containing oligosaccharides having a degree of polymerization of 1 (DP1) from the sample. In some embodiments, the detecting comprises a weight determination of at least a portion of the anhydro-subunit containing oligosaccharides having a degree of polymerization of 2 (DP2) from the sample. In some embodiments, the detecting comprises analyzing at least a portion of DP1 or DP2 anhydro-subunit containing oligosaccharides from the sample by MALDI-MS. In some embodiments, the detecting comprises analyzing at least a portion of DP1 or DP2 anhydro-subunit containing oligosaccharides from the sample by LC- MS / MS. In some embodiments, the detecting comprises analyzing at least a portion of DP1 or DP2 anhydro-subunit containing oligosaccharides from the sample by GC / MS. In some embodiments, the detecting comprises analyzing at least a portion of DP3 anhydro-subunit containing oligosaccharides from the sample. In some embodiments, the isolating comprises separating the one or more anhydro-subunit containing oligosaccharides by preparative chromatography. In some embodiments, the synthetic oligosaccharide preparation is present in the nutritional composition at a concentration of from about 1 to about 5000 ppm, from about 1 to about 1000 ppm, from about 1 to about 500 ppm, from about 10 to about 5000 ppm, from about 10 to about 2000 ppm, from about 10 to about 1000 ppm, from about 10 to about 500 ppm, from about 10 to about 250 ppm, from about 10 to about 100 ppm, from about 50 to about 5000 ppm, from about 50 to about 2000 ppm, from about 50 to about 1000 ppm, from about 50 to about 500 ppm, from about 50 to about 250 ppm, or from about 50 to about 100 ppm. In some embodiments, the synthetic oligosaccharide preparation is present in the nutritional composition at a concentration of greater than 10 ppm, greater than 50 ppm, greater than 100 ppm, greater than 200 ppm, greater than 300 ppm, greater than 400 ppm, greater than 500 ppm, greater than 600 ppm, greater than 1000 ppm, or greater than 2000 ppm. In some embodiments, the nutritional composition is an animal feed composition.

[0008] In one aspect, described herein is a method of manufacturing a nutritional composition comprising: (a) combining a base nutritional composition with a synthetic oligosaccharide preparation comprising anhydro-subunit containing oligosaccharides, and (b) performing a quality control method (e.g., methods of correlating a synthetic oligosaccharide preparation in a nutritional composition ) as provided herein. In some embodiments, the synthetic oligosaccharide preparation comprises at least n fractions of oligosaccharides each having a distinct degree of polymerization selected from 1 to n (DP1 to DPn fractions), wherein n is an integer greater than or equal to 3; and wherein the DP1 and DP2 fractions each independently comprises from about 0.5% to about 15% of anhydro-subunit containing oligosaccharides by relative abundance as measured by mass spectrometry. In some embodiments, the synthetic oligosaccharide preparation comprises at least n fractions of oligosaccharides each having a distinct degree of polymerization selected from 1 to n (DP1 to DPn fractions), wherein n is an integer greater than or equal to 2; and wherein the DP1 and DP2 fractions each independently comprises from about 0.1% to about 15% of anhydro-subunit containing oligosaccharides by relative abundance as measured by mass spectrometry. In some embodiments, the relative abundance is determined by LC-MS / MS. In some embodiments, the relative abundance of oligosaccharides in each of the n fractions decreases monotonically with its degree of polymerization.

[0009] In one aspect, provided herein is a method of quantifying an oligosaccharide preparation in a nutritional composition comprising: (a) determining a level of a signal in a sample of the nutritional composition, and (b) calculating a concentration of the oligosaccharide preparation in the nutritional composition based on the level of the signal, wherein the signal is (i) indicative of one or more anhydro-subunit containing oligosaccharides, (ii) associated with a degree of polymerization (DP) distribution of oligosaccharides, or (iii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In one aspect, provided herein is a method of performing quality control of a nutritional composition comprising: (a) detecting a signal in a sample of the nutritional composition through analytical instrumentation, and (b) accepting or rejecting a batch of the nutritional composition based on the presence or absence of the signal, wherein the signal is (i) indicative of one or more anhydro-subunit containing oligosaccharides, (ii) associated with a degree of polymerization (DP) distribution of oligosaccharides, or (iii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>- <semantics>(1,1)<annotation encoding="application / x-tex">(1,1)< / annotation>< / semantics>-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-<semantics>(1,1)<annotation encoding="application / x-tex">(1,1)< / annotation>< / semantics>-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-<semantics>(1,1)<annotation encoding="application / x-tex">(1,1)< / annotation>< / semantics>-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- <semantics>(1,1)<annotation encoding="application / x-tex">(1,1)< / annotation>< / semantics>-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In another aspect, provided herein is a method of performing quality control of a nutritional composition comprising: (a) detecting, through analytical instrumentation, the presence or absence of a first signal in a first sample of the nutritional composition, and a second signal in a second sample of the nutritional composition, and (b) comparing the first signal and the second signal, wherein the first signal and the second signal are (i) indicative of one or more anhydro-subunit containing oligosaccharides, (ii) associated with a degree of polymerization (DP) distribution of oligosaccharides, or (iii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In some embodiments, the signal, the level of the signal, the first signal, and / or the second signal are determined or detected by high-performance liquid chromatography (HPLC), gas chromatography (GC), mass spectrometry (MS), nuclear magnetic resonance (NMR) spectroscopy, size exclusion chromatography (SEC), field flow fractionation (FFF), asymmetric flow field flow fractionation (A4F), or any combination thereof. In some embodiments, the nutritional composition comprises a base nutritional composition. In some embodiments, the signal, first signal, and / or the second signal is indicative of one or more anhydro-subunit containing oligosaccharides. In some embodiments, the one or more anhydro-subunit containing oligosaccharides originate from the oligosaccharide preparation. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to anhydro-subunit containing oligosaccharides in the DP1 fraction. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to levoglucosan, 1,6-anhydro-β-D-glucofuranose, or a combination thereof. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to one or more anhydro-subunit containing oligosaccharides in the DP2 fraction. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to cellobiosan. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to anhydro- subunit containing oligosaccharides in the DP3 fraction. In some embodiments, the first signal and the second signal are attributed to the same species of an anhydro-subunit containing oligosaccharide. In some embodiments, the first signal and the second signal are attributed to different species of anhydro-subunit containing oligosaccharides. In some embodiments, the signal, the level of the signal, the first signal, and / or the second signal are determined or detected by matrix-assisted laser desorption / ionization-mass spectrometry (MALDI-MS). In some embodiments, the signal, the level of the signal, the first signal, and / or the second signal are determined or detected by liquid chromatography—mass spectrometry (LC-MS) / MS. In some embodiments, the signal, the level of the signal, the first signal, and / or the second signal are determined or detected by GC-flame ionization detector (GC-FID) or GC-MS. In some embodiments, the signal, the level of the signal, the first signal, and / or the second signal are determined or detected by NMR. In some embodiments, a derivatization step is performed prior to detection. In some embodiments, the signal, the level of the signal, the first signal, and / or the second signal are determined by the weight of isolated and / or purified fractions from preparative chromatography. In some embodiments, the base nutritional composition lacks a detectable level of anhydro-subunits. In some embodiments, the base nutritional composition is essentially free of anhydro-subunits. In certain embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In some embodiments, the signal, the first signal, and / or the second signal are detected or determined by NMR. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages. In certain embodiments, the signal, the first signal, and / or the second signal are associated with a DP distribution of oligosaccharides. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to the oligosaccharides in the DP2 fraction. In some embodiments, the method further comprises accepting or rejecting a batch of the nutritional composition. In some embodiments, the method further comprises adjusting the level of the oligosaccharide preparation after the determining or detecting. In some embodiments, the base nutritional composition comprises a plurality of oligosaccharides. In some embodiments, the base nutritional composition comprises starch and / or plant fibers. In some embodiments, the level of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkage, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkage, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkage in the base nutritional composition is at least 10% lower than the level of the same glycosidic linkage in the oligosaccharide preparation. In some embodiments, the level of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)- α glycosidic linkages, α-(1,1)-β glycosidic linkages, β-(1,1)-α glycosidic linkages, or β-(1,1)-β glycosidic linkages of oligosaccharides in the base nutritional composition is at least 10% lower than the level of the same glycosidic linkage in the oligosaccharide preparation. In some embodiments, the method further comprises extracting oligosaccharides from a sample of the nutritional composition. In some embodiments, the method further comprises filtering or clarifying the extracted oligosaccharides. In some embodiments, the method further comprises concentrating the extracted oligosaccharides. In some embodiments, the concentrating the extracted oligosaccharides comprises freeze-drying. In some embodiments, the method further comprises introducing an internal standard into the extracted or concentrated oligosaccharides. In some embodiments, the method further comprises reducing the extracted or concentrated oligosaccharides. In some embodiments, the method further comprises digesting the extracted or concentrated oligosaccharides with one or more hydrolytic enzymes. In some embodiments, the one or more hydrolytic enzymes comprise carbohydratase, protease, lipase, or any combination thereof. In some embodiments, the one or more hydrolytic enzymes comprise <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-amylase, amyloglycosidase, invertase, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-galactosidase, or any combination thereof. In some embodiments, the one or more hydrolytic enzymes cleave one or more naturally occurring glycosidic bonds. In some embodiments, the method further comprises isolating the oligosaccharides that are not digested. In some embodiments, the method further comprises separating the extracted, concentrated, digested, or reduced oligosaccharides. In some embodiments, the oligosaccharides are separated chromatographically. In some embodiments, the method further comprises isolating the separated oligosaccharides. In some embodiments, the oligosaccharides are separated or isolated by the degree of polymerization. In some embodiments, the method comprises isolating or separating the oligosaccharides with a degree of polymerization of 1, 2, 3, 4, or 5. In some embodiments, the oligosaccharides in the DP1 fraction are isolated or separated. In some embodiments, the oligosaccharides in the DP2 fraction are isolated or separated. In some embodiments, the oligosaccharides in the DP3 fraction are isolated or separated. In some embodiments, the oligosaccharides in the DP4 fraction are isolated or separated. In some embodiments, the oligosaccharides in the DP5 fraction are isolated or separated. In some embodiments, the isolated or separated oligosaccharides are quantified. In some embodiments, the anhydro-subunit containing oligosaccharides within the isolated or separated oligosaccharides are quantified. In some embodiments, the majority of the quantified oligosaccharides are from the oligosaccharide preparation. In some embodiments, more than 50, 60, 70, 80, 90, 95, or 99 wt% of the quantified oligosaccharide are from the oligosaccharide preparation. In some embodiments, the method comprises analyzing glycosidic linkages of the oligosaccharides by NMR, wherein the glycosidic linkages are <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, thereby determining or detecting the signal, the level of the signal, the first signal, or the second signal associated with the corresponding glycosidic linkages. In some embodiments, the method comprises analyzing the anhydro-subunit containing oligosaccharides by mass spectrometry, thereby determining or detecting the signal, the level of the signal, the first signal, or the second signal indicative of one or more anhydro- subunit containing oligosaccharides. In some embodiments, the method comprises analyzing the anhydro-subunit containing oligosaccharides by HPLC, thereby determining or detecting the signal, the level of the signal, the first signal, or the second signal indicative of one or more anhydro-subunit containing oligosaccharides. In some embodiments, the method comprises analyzing the anhydro-subunit containing oligosaccharides by FFF or A4F, thereby determining or detecting the signal, the level of the signal, the first signal, or the second signal indicative of one or more anhydro-subunit containing oligosaccharides. In some embodiments, the method comprises analyzing the DP distribution of the oligosaccharides by SEC, GC, or HPLC, thereby determining or detecting the signal, the level of the signal, the first signal, or the second signal associated with the DP distribution. In some embodiments, the method comprises quantifying the oligosaccharides in the DP2 fraction by SEC, GC, or HPLC, thereby determining or detecting the signal, the level of the signal, the first signal, or the second signal associated with the DP distribution. In some embodiments, the oligosaccharide preparation is from 1 to 5000 ppm, 1 to 1000 ppm, 1 to 500 ppm, 10 to 5000 ppm, 10 to 2000 ppm, 10 to 1000 ppm, 10 to 500 ppm, 10 to 250 ppm, 10 to 100 ppm, 50 to 5000 ppm, 50 to 2000 ppm, 50 to 1000 ppm, 50 to 500 ppm, 50 to 250 ppm, or 50 to 100 ppm relative to the nutritional composition. In some embodiments, the oligosaccharide preparation is greater than 10 ppm, 50 ppm, 100 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm, 1000 ppm, or 2000 ppm relative to the nutritional composition. In some embodiments, the nutritional composition is an animal feed composition.

[0010] In one aspect, provided herein is a method of manufacturing a nutritional composition comprising: (a) combining a synthetic oligosaccharide preparation comprising anhydro-subunit containing oligosaccharides with a base nutritional composition, and (b) performing a quality control step as provided herein. In some embodiments, the synthetic oligosaccharide preparation comprises at least n fractions of oligosaccharides each having a distinct degree of polymerization selected from 1 to n (DP1 to DPn fractions), wherein n is an integer greater than or equal to 2; wherein each fraction comprises from 0.1% to 15% anhydro-subunit containing oligosaccharides by relative abundance as measured by mass spectrometry. In some embodiments, the synthetic oligosaccharide preparation comprises at least n fractions of oligosaccharides each having a distinct degree of polymerization selected from 1 to n (DP1 to DPn fractions), wherein n is an integer greater than or equal to 3; wherein each fraction comprises from 0.5% to 15% anhydro- subunit containing oligosaccharides by relative abundance as measured by mass spectrometry. In some embodiments, the relative abundance of oligosaccharides in each of the n fractions decreases monotonically with its degree of polymerization.

[0011] Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in this art from the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various obvious respects, all without departing from the disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive.

[0012] To the extent publications and patents or patent applications referred to herein contradict the disclosure contained in the specification, the specification is intended to supersede and / or take precedence over any such contradictory material. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawing (also "figure" and "FIG." herein), of which:

[0014] FIG. 1 illustrates a 1H, 13C- HSQC NMR spectrum of the oligosaccharide preparation of Example 9.7.

[0015] FIG. 2 illustrates an MALDI-MS spectrum of an oligosaccharide preparation from Example 9 that demonstrates the presence of anhydro-subunits.

[0016] FIG. 3 illustrates a 1D 1H- proton NMR spectrum of an anhydro DP1 fraction isolated from an oligosaccharide of Example 9.

[0017] FIG. 4 illustrates a 1D APT 13C- NMR spectrum of an anhydro DP1 fraction isolated from an oligosaccharide of Example 9.

[0018] FIG. 5 illustrates the structures of two anhydro DP1 compounds (1,6-anhydro-beta-D- glucofuranose and 1,6-anhydro-beta-D-glucopyranose) and their NMR assignments.

[0019] FIG. 6 illustrates an enlargement of the GC-MS chromatogram (TIC and XIC (m / z 229) plots) for the oligosaccharide preparation of Example 9.7 following derivatization.

[0020] FIG. 7 illustrates MALDI-MS spectra comparing the oligosaccharide preparation from Example 9 versus a conventional dextran.

[0021] FIG. 8 illustrates LC-MS / MS detection of the anhydro DP2 species at concentration of 1 - 80 μg / mL of an oligosaccharide preparation in water.

[0022] FIG. 9 illustrates a linear calibration curve resulting from the LC-MS / MS detection of FIG. 8.

[0023] FIG. 10 illustrates the quantification of the anhdro-DP2 content of various control and treated diet compositions.

[0024] FIG. 11 illustrates a 2D-1H JRES NMR spectrum of an anhydro-subunit containing gluco- oligosaccharides sample.

[0025] FIG. 12 is a representative 1H, 13C- HSQC NMR spectrum of an anhydro-subunit containing gluco-oligosaccharides sample with relevant resonances and assignments used for linkage distribution.

[0026] FIG. 13 illustrates an overlay of 1H DOSY spectra of three anhydro-subunit containing oligosaccharides.

[0027] FIG. 14 illustrates a comparison of 1,6-Anhydro-\(\beta\)-D-glucose (DP1-18), 1,6-Anhydro-\(\beta\)- D-cellobiose (DP2-18), and an anhydro-subunit containing oligosaccharides sample.

[0028] FIG. 15 illustrates mass chromatograms of anhydro-subunit containing oligosaccharides (top) and digested anhydro-subunit containing oligosaccharides (bottom) at selected multiple reaction monitoring (MRM).

[0029] FIG. 16 illustrates mass chromatograms of (1) feed that contains anhydro-subunit containing oligosaccharides, (2) digested feed that contains anhydro-subunit containing oligosaccharides, and (3) blank digested feed at selected MRM.

[0030] FIG. 17 illustrates a representative workflow for the analysis of oligosaccharide preparations in animal feed.

[0031] FIG. 18 illustrates two DP1 and one DP2 anhydro-subunit containing oligosaccharides.

[0032] FIG. 19 illustrates an anhydro-subunit containing oligosaccharide (cellotriosan).

[0033] FIG. 20A illustrates a MALDI-MS spectrum of an oligosaccharide preparation from Example 2 that demonstrates the presence of anhydro-subunits; FIG. 20B illustrates an enlargement of a part of the MALDI-MS spectrum shown in FIG. 20A.

[0034] FIG. 21A illustrates LC-MS / MS detection of the anhydro DP2 species of an oligosaccharide preparation of Example 1; FIG. 21B illustrates LC-MS / MS detection of the anhydro DP1 species of an oligosaccharide preparation of Example 1; FIG. 21C illustrates LC- MS / MS detection of the DP2 species of an oligosaccharide preparation of Example 1.

[0035] FIG. 22A illustrates LC-MS / MS detection of the anhydro DP2 species of an oligosaccharide preparation of Example 3; FIG. 22B illustrates LC-MS / MS detection of the anhydro DP1 species of an oligosaccharide preparation of Example 3; FIG. 22C illustrates LC- MS / MS detection of the DP2 species of an oligosaccharide preparation of Example 3.

[0036] FIG. 23A illustrates LC-MS / MS detection of the anhydro DP2 species of an oligosaccharide preparation of Example 4; FIG. 23B illustrates LC-MS / MS detection of the anhydro DP1 species of an oligosaccharide preparation of Example 4; FIG. 23C illustrates LC- MS / MS detection of the DP2 species of an oligosaccharide preparation of Example 4.

[0037] FIG. 24A illustrates LC-MS / MS detection of the anhydro DP2 species of an oligosaccharide preparation of Example 7; FIG. 24B illustrates LC-MS / MS detection of the anhydro DP1 species of an oligosaccharide preparation of Example 7; FIG. 24C illustrates LC- MS / MS detection of the DP2 species of an oligosaccharide preparation of Example 7.

[0038] FIG. 25A illustrates GC-MS spectrum detection of the DP1, anhydro DP1, DP2 and anhydro DP2 fractions of an oligosaccharide preparation of Example 1; FIG. 25B illustrates an enlargement of the DP2 and anhydro DP 2 fractions as shown in FIG. 25A.

[0039] FIG. 26A illustrates GC-MS spectrum detection of the DP1, anhydro DP1, DP2 and anhydro DP2 fractions of an oligosaccharide preparation of Example 3; FIG. 26B illustrates an enlargement of the DP2 and anhydro DP 2 fractions as shown in FIG. 26A.

[0040] FIG. 27A illustrates GC-MS spectrum detection of the DP1, anhydro DP1, DP2 and anhydro DP2 fractions of an oligosaccharide preparation of Example 4; FIG. 27B illustrates an enlargement of the DP2 and anhydro DP 2 fractions as shown in FIG. 27A.

[0041] FIG. 28A illustrates GC-MS spectrum detection of the DP1, anhydro DP1, DP2 and anhydro DP2 fractions of an oligosaccharide preparation of Example 7; FIG. 28B illustrates an enlargement of the DP2 and anhydro DP 2 fractions as shown in FIG. 28A.

[0042] FIG. 29 illustrates the effect of reaction temperature, water content, and reaction time on the content of DP2 anhydro-subunit containing oligosaccharides in the oligosaccharide preparations, as compared to an oligosaccharide preparation according to Example 2.

[0043] FIG. 30 illustrates MALDI-MS spectra comparing the oligosaccharide preparation from Example 9 at different laser energies. DETAILED DESCRIPTION

[0044] The following description and examples illustrate embodiments of the present disclosure in detail. It is to be understood that this present disclosure is not limited to the particular embodiments described herein and as such can vary. Those of skill in the art will recognize that there are numerous variations and modifications of this present disclosure, which are encompassed within its scope.

[0045] All terms are intended to be understood as they would be understood by a person skilled in the art. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosure pertains.

[0046] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0047] Although various features of the present disclosure may be described in the context of a single embodiment, the features may also be provided separately or in any suitable combination. Conversely, although the present disclosure may be described herein in the context of separate embodiments for clarity, the present disclosure may also be implemented in a single embodiment.

[0048] The following definitions supplement those in the art and are directed to the current application and are not to be imputed to any related or unrelated case, e.g., to any commonly owned patent or application. Although any methods and materials similar or equivalent to those described herein can be used in the practice for testing of the present disclosure, the preferred materials and methods are described herein. Accordingly, the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting. I. Definitions

[0049] As used herein the term "administering" includes providing a synthetic oligosaccharide preparation, a nutritional composition, a liquid, or an animal feed composition described herein, to an animal such that the animal may ingest the synthetic oligosaccharide preparation, the nutritional composition, the liquid, or the animal feed composition. In such embodiments, the animal ingests some portion of the synthetic oligosaccharide preparation, the nutritional composition, or the animal feed composition. In some embodiments, the animal ingests some portion of the synthetic oligosaccharide preparation, the nutritional composition, the liquid, or the animal feed composition in every 24-hour period or every other 24-hour period for at least 7 days, 14 days, 21 days, 30 days, 45 days, 60 days, 75 days, 90 days or 120 days. In some embodiments, the oligosaccharide preparation may be dissolved in water or another liquid, and the animal ingests some portion of the oligosaccharide preparation by drinking the liquid. In certain embodiments, the oligosaccharide is provided to the animal via its drinking water. In certain embodiments, the oligosaccharide preparation, nutritional composition, liquid, or animal feed composition is consumed at will.

[0050] As used herein, the term "inclusion level" or "dose" refers to the concentration of an oligosaccharide preparation in a nutritional composition, a liquid, a diet, or an animal feed composition provided to the animal. In some embodiments, the inclusion level is measured as the mass concentration of the oligosaccharide preparation in the final nutritional composition, liquid, diet, or animal feed. For example, the inclusion level may be measured in units of parts per million (ppm) of the oligosaccharide on a dry solids weight basis per the total weight of the final nutritional composition, liquid, diet, or animal feed. In certain embodiments, the dry solids mass of the oligosaccharide preparation is measured as the dry-basis mass of DP1+ species. In other embodiments, the dry solids mass of the oligosaccharide preparation is measured as the dry-basis mass of DP2+ species.

[0051] As used herein, the term "specific dose" refers to the quantity of an oligosaccharide preparation consumed by an animal per unit of time and relative to its body mass. In some embodiments, the specific dose may be measured in units of mg of oligosaccharide preparation (on a dry solids-basis) per kg of body weight of the animal per day (i.e., mg / kg / day).

[0052] As used herein, the term "anhydro-subunit" refers to a product of thermal dehydration of a monosaccharide (or monosaccharide subunit) or a sugar caramelization product. For example, an "anhydro-subunit" can be an anhydro-monosaccharide such as anhydro-glucose. As another example, an "anhydro-subunit" can be linked with one or more regular or anhydro- monosaccharide subunits via glycosidic linkage.

[0053] As used herein, the term an "anhydro DPn oligosaccharide," an "anhydro DPn species," or a "DPn anhydro-subunit containing oligosaccharide" refers to an oligosaccharide that has a degree of polymerization of n and comprises one or more anhydro-subunits. As such, an anhydro- glucose is a DP1 anhydro-subunit containing oligosaccharide and an cellotriosan is a DP3 anhydro-subunit containing oligosaccharide.

[0054] As used herein the term "feed conversion ratio (FCR)," refers to the ratio of feed mass input (for example consumed by the animal) to the animal output, wherein the animal output is the target animal product. For example, the animal output for dairy animals is milk, whereas the animal output for animals raised for meat is body mass.

[0055] The term "oligosaccharide" refers to a monosaccharide or a compound containing two or more monosaccharide subunits linked by glycosidic bonds. As such, an oligosaccharide includes a regular monosaccharide; an anhydro-monosaccharide; or a compound containing two or more monosaccharide subunits, wherein one or more monosaccharide subunits are optionally, independently replaced by one or more anhydro-subunits. An oligosaccharide can be functionalized. As used herein, the term oligosaccharide encompasses all species of the oligosaccharide, wherein each of the monosaccharide subunit in the oligosaccharide is independently and optionally functionalized and / or replaced with its corresponding anhydro- monosaccharide subunit.

[0056] As used herein, the term "oligosaccharide preparation" refers to a preparation that comprises at least one oligosaccharide.

[0057] As used herein, the term "gluco-oligosaccharide" refers to a glucose or a compound containing two or more glucose monosaccharide subunits linked by glycosidic bonds. As such, a gluco-oligosaccharide includes a glucose; an anhydro-glucose; or a compound containing two or more glucose monosaccharide subunits linked by glycosidic bonds, wherein one or more of said glucose monosaccharide subunits are each optionally and independently replaced with an anhydro-glucose subunit.

[0058] As used herein, the term "galacto-oligosaccharide" refers to a galactose or a compound containing two or more galactose monosaccharide subunits linked by glycosidic bonds. As such, a galacto-oligosaccharide includes a galactose; an anhydro-galactose or a compound containing two or more galactose monosaccharide subunits linked by glycosidic bonds, wherein one or more of said galactose monosaccharide subunits are each optionally and independently replaced with an anhydro-galactose subunit.

[0059] As used herein, the term "gluco-galacto-oligosaccharide preparation" refers to a composition that is produced from a complete or incomplete sugar condensation reaction of glucose and galactose. Accordingly, in some embodiments, a gluco-galactose-oligosaccharide preparation comprises gluco-oligosaccharides, galacto-oligosaccharides, compounds containing one or more glucose monosaccharide subunits and one or more galactose monosaccharide subunits linked by glycosidic bonds, or a combination thereof. In some embodiments, a gluco-galactose- oligosaccharide preparation comprises gluco-oligosaccharides and compounds containing one or more glucose monosaccharide subunits and one or more galactose monosaccharide subunits linked by glycosidic bonds. In some embodiments, a gluco-galactose-oligosaccharide preparation comprises galacto-oligosaccharides and compounds containing one or more glucose monosaccharide subunits and one or more galactose monosaccharide subunits linked by glycosidic bonds. In some embodiments, a gluco-galactose-oligosaccharide preparation comprises compounds containing one or more glucose monosaccharide subunits and one or more galactose monosaccharide subunits linked by glycosidic bonds.

[0060] As used herein, the term "monosaccharide unit" and "monosaccharide subunit" are used interchangeably. A "monosaccharide subunit" refers to a monosaccharide monomer in an oligosaccharide. For an oligosaccharide having a degree of polymerization of 1, the oligosaccharide can be referred to as a monosaccharide subunit or monosaccharide. For an oligosaccharide having a degree of polymerization of 2 or higher, its monosaccharide subunits are linked via glycosidic bonds.

[0061] As used herein, the term "regular monosaccharide" refers to a monosaccharide that does not contain an anhydro-subunit. The term "regular disaccharide" refers to a disaccharide that does not contain an anhydro-subunit. Accordingly, the term "regular subunit" refers to a subunit that is not an anhydro-subunit.

[0062] The term "relative abundance" or "abundance," as used herein, refers to the abundance of a species in terms of how common or rare the species exists. For example, a DP1 fraction comprising 10% anhydro-subunit containing oligosaccharides by relative abundance can refer to a plurality of DP1 oligosaccharides, wherein 10% of the DP1 oligosaccharides are anhydro- monosaccharides. The relative abundance, e.g., for a certain DP fraction of oligosaccharides, can be determined by suitable analytical instrumentations, for example, mass spectrometry and liquid chromatography such as LC-MS / MS, GC-MS, HPLC-MS, and MALDI-MS. In some embodiments, the relative abundance is determined by integrating the area under the peaks of the chromatographs (e.g., LC-MS / MS, GC-MS, and HPLC-MS) that correspond to the fractions of interest. In some embodiments, the relative abundance is determined by the peak intensities (e.g., MALDI-MS). In some embodiments, the relative abundance is determined by a combination of analytical methods such as a weight determination after separation by liquid chromatography.

[0063] As used herein, the singular forms "a," "and," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "an agent" includes a plurality of such agents, and reference to "the oligosaccharide" includes reference to one or more oligosaccharides (or to a plurality of oligosaccharides) and equivalents thereof known to those skilled in the art, and so forth.

[0064] When ranges are used herein for physical properties, such as molecular weight, or chemical properties, such as chemical formulae, all combinations and subcombinations of ranges and specific embodiments therein are intended to be included. The term "about" when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range, in some instances, will vary between 1% and 15% of the stated number or numerical range. In some embodiments, the term "about" means within 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, or 0.05% of a given value or range.

[0065] The term "comprising" (and related terms such as "comprise" or "comprises" or "having" or "including") is intended to include, but not necessarily be limited to the things so described. II. Applications of the Methods

[0066] Oligosaccharide preparations are used as additives in nutritional compositions such as complete animal feed. The addition of such oligosaccharide preparations improves the health and performance (weight, growth rate, feed conversion efficiency) of animals such as poultry (broilers, layers, broiler breeders, turkeys) and pigs (nursery pigs, grower / finisher pigs, sows, etc., and other species).

[0067] The impact of the oligosaccharide preparation on animal health and performance depends on the chemical and physiochemical properties of the oligosaccharides. In particular, the chemical composition of the oligosaccharide preparation can affect the manner in which it is utilized by the microflora of the animal, in particular by the gut microbiome. The microflora of the animal, and in particular the gut microbiome, in turn affects the health and performance of the animal as a whole. Such effects include, for example, physiological and morphological effects on the digestive tract, immune system activation, stimulation of mucus production, improvement of the gut barrier function, improvement in the liberation and absorption of nutrients, modulation of the abundance of the various members of the gut microflora (e.g., suppression of pathogenic species, augmentation of beneficial commensal species), and modulation of biochemical species produced by the gut microflora. Accordingly, oligosaccharide preparations can be added to nutritional compositions such as animal feed to act as prebiotics.

[0068] The method of producing oligosaccharide preparations suitable for use as an additive in nutritional compositions are provided herein. Oligosaccharide preparations can also be produced by other means, including enzymatic hydrolysis or acid hydrolysis of plant fiber, extraction of bacterial and yeast cell wall glycans and glycopeptides, enzymatic condensation from saccharides, and fermentation by wild type or recombinant micro-organisms.

[0069] Due to the respective structural complexities of both the nutritional compositions and the oligosaccharide preparations, a simple, selective, and sensitive technique for analyzing the presence and concentration of oligosaccharide preparations within the feed material is lacking. The nutritional compositions comprise a large quantity and diversity of carbohydrate structures (e.g., starch, plant fibers and pectins). It is therefore particularly challenging to distinguish small quantities of oligosaccharide-based feed additives from the vast sea of other carbohydrates present as base of the nutritional composition. As such, an analytical method with the required sensitivity and selectivity is needed to distinguish such oligosaccharide feed additives from natural oligomers.

[0070] It is of commercial utility to assay for the presence and / or concentration of feed additives. Such assay can be performed for the purpose of quality control, to determine whether the additive was blended consistently with the base nutritional composition to provide a final nutritional composition comprising the additive at the intended dose or level of inclusion. In certain embodiments, the manufacturing process can include a step of either accepting or rejecting a quantity (e.g., a lot or batch) of the final feed composition based on determining that the additive was included within a range of a pre-determined value.

[0071] Such oligosaccharide preparations comprise a diversity of glycosidic linkages between the various monomers in the oligosaccharide preparation. In certain embodiments, the relative abundances of glycosidic linkages, e.g., <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) linkages, easily hydrolyzed by the digestive acids and enzymes of the animal's upper digestive are low. As such, the oligosaccharides survive primary digestion by the animal and proceed through the digestive track to the lower digestive track where they interact with the gut microbiome. In some embodiments, the relative abundances of glycosidic linkages commonly encountered in plant fibers and pectins are also low. In some embodiments, the oligosaccharide preparations comprise linkages that occur only with low relative abundance in the base carbohydrates (i.e., the background carbohydrates) in the nutritional composition. In certain embodiments, the oligosaccharide preparations comprise anhydro-subunit containing oligosaccharides. In certain embodiments, the oligosaccharide preparations comprise chemically synthesized anhydro-subunit containing oligosaccharides.

[0072] Despite the need for detecting and / or quantifying the relative abundance of the oligosaccharide preparations in nutritional compositions, existing analytical methods for quantifying various glycosidic linkages in complex carbohydrates can lack sensitivity. Certain existing analytical methods have a minimum threshold of resolving glycosidic linkages at a level of several percentages. Commercially relevant inclusion levels of feed additives are, however, typically in the range of 1 to 5000 ppm, 10 to 1000 ppm, 10 to 500 ppm, or 50 to 500 ppm. Thus, the glycosidic linkages in feed additives are not detectable via some of the existing methods.

[0073] Surprisingly, we have found that oligosaccharide preparations can be analyzed and quantified in a complex nutritional composition such as complete animal feed as provided herein. III. Oligosaccharide Preparations Oligosaccharides by Manufacturing Method

[0074] Provided herein are oligosaccharide preparations suitable for use in nutritional compositions. In some embodiments, oligosaccharide preparations, as provided herein, can comprise monosaccharides, oligosaccharides, polysaccharides, or any combination thereof, wherein one or more monosaccharide subunits in any of the monosaccharides, oligosaccharides, or polysaccharides can be independently functionalized. In some embodiments, the oligosaccharide preparations comprise oligosaccharides produced by hydrolysis or pyrolysis of polysaccharides such as cellulose and starch, by condensation or polymerization of monosaccharides or oligosaccharides, by enzymatic hydrolysis or acid hydrolysis of polysaccharides such as plant fiber, by extraction of bacterial and yeast cell wall glycans and glycopeptides, by enzymatic condensation from saccharides, by fermentation by wild type or recombinant micro-organisms, or by any combination thereof. In some embodiments, the oligosaccharide preparations can comprise any oligosaccharides known in the art. In some embodiments, the oligosaccharide preparations are produced chemically, naturally, or enzymatically.

[0075] In one aspect, a described oligosaccharide preparation suitable for the methods described herein is a synthetic oligosaccharide preparation. In some embodiments, a synthetic oligosaccharide preparation refers to a plurality of oligosaccharides produced by a process that does not require live organisms. In some embodiments, a synthetic oligosaccharide preparation refers to a plurality of oligosaccharides produced by a process that does not require enzymes. In some embodiments, a synthetic oligosaccharide preparation refers to a plurality of oligosaccharides produced by a chemical process. In certain embodiments, a synthetic oligosaccharide preparation refers to a plurality of oligosaccharides produced by the condensation of sugars. Degree of Polymerization (DP) Distribution

[0076] In some embodiments, a herein described oligosaccharide preparation comprises at least n fractions of oligosaccharides each having a distinct degree of polymerization selected from 1 to n (DP1 to DPn fractions). In some embodiments, the oligosaccharide preparation comprises n fractions of oligosaccharides, each fraction having a distinct degree of polymerization selected from 1 to n (DP1 to DPn fractions). In some embodiments, the DP1 fraction comprises one or more monosaccharides and / or one or more anhydro-monosaccharides. As another example, in some embodiments, the DP1 fraction comprises glucose, galactose, fructose, 1,6-anhydro-β-D- glucofuranose, 1,6-anhydro-β-D-glucopyranose, or any combination thereof. For example, in some embodiments, the DP2 fraction comprises one or more regular disaccharides and one or more anhydro-subunit containing disaccharides. In some embodiments, the DP2 fraction comprises lactose.

[0077] In some embodiments, n is at least 2, at least 3, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45, at least 46, at least 47, at least 48, at least 49, at least 50, at least 51, at least 52, at least 53, at least 54, at least 55, at least 56, at least 57, at least 58, at least 59, at least 60, at least 61, at least 62, at least 63, at least 64, at least 65, at least 66, at least 67, at least 68, at least 69, at least 70, at least 71, at least 72, at least 73, at least 74, at least 75, at least 76, at least 77, at least 78, at least 79, at least 80, at least 81, at least 82, at least 83, at least 84, at least 85, at least 86, at least 87, at least 88, at least 89, at least 90, at least 91, at least 92, at least 93, at least 94, at least 95, at least 96, at least 97, at least 98, at least 99, or at least 100. In some embodiments, n is 2, 3, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100. In some embodiments, n is less than 10, less than 11, less than 12, less than 13, less than 14, less than 15, less than 16, less than 17, less than 18, less than 19, less than 20, less than 21, less than 22, less than 23, less than 24, less than 25, less than 26, less than 27, less than 28, less than 29, less than 30, less than 31, less than 32, less than 33, less than 34, less than 35, less than 36, less than 37, less than 38, less than 39, less than 40, less than 41, less than 42, less than 43, less than 44, less than 45, less than 46, less than 47, less than 48, less than 49, less than 50, less than 51, less than 52, less than 53, less than 54, less than 55, less than 56, less than 57, less than 58, less than 59, less than 60, less than 61, less than 62, less than 63, less than 64, less than 65, less than 66, less than 67, less than 68, less than 69, less than 70, less than 71, less than 72, less than 73, less than 74, less than 75, less than 76, less than 77, less than 78, less than 79, less than 80, less than 81, less than 82, less than 83, less than 84, less than 85, less than 86, less than 87, less than 88, less than 89, less than 90, less than 91, less than 92, less than 93, less than 94, less than 95, less than 96, less than 97, less than 98, less than 99, or less than 100. In some embodiments, n is from 2 to 100, from 5 to 90, from 10 to 90, from 10 to 80, from 10 to 70, from 10 to 60, from 10 to 50, from 10 to 40, from 10 to 30, from 15 to 60, from 15 to 50, from 15 to 45, from 15 to 40, from 15 to 35, or from 15 to 30.

[0078] A distribution of the degree of polymerization of the oligosaccharide preparation can be determined by any suitable analytical method and instrumentation, including but not limited to end group method, osmotic pressure (osmometry), ultracentrifugation, viscosity measurements, light scattering method, size exclusion chromatography (SEC), SEC-MALLS, field flow fractionation (FFF), asymmetric flow field flow fractionation (A4F), high-performance liquid chromatography (HPLC), and mass spectrometry (MS). For example, the distribution of the degree of polymerization can be determined and / or detected by mass spectrometry, such as matrix- assisted laser desorption / ionization (MALDI)-MS, liquid chromatography (LC)-MS, or gas chromatography (GC)-MS. For another example, the distribution of the degree of polymerization can be determined and / or detected by SEC, such as gel permeation chromatography (GPC). As yet another example, the distribution of the degree of polymerization can be determined and / or detected by HPLC, FFF, or A4F. In some embodiments, the distribution of the degree of polymerization is determined and / or detected by MALDI-MS. In some embodiments, the distribution of the degree of polymerization is determined and / or detected by GC-MS or LC-MS. In some embodiments, the distribution of the degree of polymerization is determined and / or detected by SEC. In some embodiments, the distribution of the degree of polymerization is determined and / or detected by HPLC. In some embodiments, the distribution of the degree of polymerization is determined and / or detected by a combination of analytical instrumentations such as MALDI-MS and SEC. In some embodiments, the degree of polymerization of the oligosaccharide preparation can be determined based on its molecular weight and molecular weight distribution. For example, FIG. 2 shows a MALDI-MS spectrum that illustrates the degrees of polymerizations of various fractions and the presence of anhydro-subunit containing oligosaccharides (the -18 g / mol MW offset peaks) in all of the observed fractions.

[0079] In some embodiments, the relative abundance of oligosaccharides in a majority of the fractions decreases monotonically with its degree of polymerization. In some embodiments, the relative abundance of oligosaccharides of less than 6, less than 5, less than 4, less than 3, or less than 2 fractions of the oligosaccharide preparation do not decrease monotonically with its degree of polymerization.

[0080] In some embodiments, the relative abundance of oligosaccharides in at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50 DP fractions decreases monotonically with its degree of polymerization. In some embodiments, the relative abundance of oligosaccharides in at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50 consecutive DP fractions decreases monotonically with its degree of polymerization. In some embodiments, the relative abundance of oligosaccharides in at least 5, at least 10, at least 20, or at least 30 DP fractions decreases monotonically with its degree of polymerization. In some embodiments, the relative abundance of oligosaccharides in at least 5, at least 10, at least 20, or at least 30 consecutive DP fractions decreases monotonically with its degree of polymerization.

[0081] In some embodiments, the relative abundance of oligosaccharides in each of the n fractions decreases monotonically with its degree of polymerization. For example, FIG. 10 provides an example of a DP distribution where the relative abundance of oligosaccharides in each of the n fractions decrease monotonically with its DP. For example, in some embodiments, only the relative abundance of oligosaccharides in the DP3 fraction does not decrease monotonically with its degree of polymerization, i.e., the relative abundance of oligosaccharides in the DP3 fraction is lower than the relative abundance of oligosaccharides in the DP4 fraction. In some embodiments, the relative abundance of oligosaccharides in the DP2 fraction is lower than the relative abundance of oligosaccharides in the DP3 fraction.

[0082] In some embodiments, a herein described oligosaccharide preparation has a DP1 fraction content of from about 1% to about 50%, from about 1% to about 40%, from about 1% to about 35%, from about 1% to about 30%, from about 1% to about 25%, from about 1% to about 20%, from about 1% to about 15%, from about 5% to about 50%, from about 5% to about 40%, from about 5% to about 35%, from about 5% to about 30%, from about 5% to about 25%, from about 5% to about 20%, from about 5% to about 15%, from about 10% to about 50%, from about 10% to about 40%, from about 10% to about 35%, from about 10% to about 30%, from about 10% to about 25%, from about 10% to about 20%, or from about 10% to about 15% by weight or by relative abundance. In some embodiments, the oligosaccharide preparation has a DP1 fraction content of from about 10% to about 35%, from about 10% to about 20%, or from about 10% to about 15% by weight or by relative abundance. In some embodiments, the content of the DP1 fraction is determined by mass spectrometry. In some embodiments, the content of the DP1 fraction is determined by HPLC. In some embodiments, the content of the DP1 fraction is determined by LC-MS / MS or GC-MS.

[0083] In some embodiments, a herein described oligosaccharide preparation has a DP2 fraction content of from about 1% to about 35%, from about 1% to about 30%, from about 1% to about 25%, from about 1% to about 20%, from about 1% to about 15%, from about 1% to about 10%, from about 5% to about 30%, from about 5% to about 25%, from about 5% to about 20%, from about 5% to about 15%, or from about 5% to about 10% by weight or by relative abundance. In some embodiments, the oligosaccharide preparation has a DP2 fraction content of from about 5% to about 25%, from about 5% to about 20%, from about 5% to about 15%, or from about 5% to about 10% by weight or by relative abundance. In some embodiments, the content of the DP2 fraction is determined by mass spectrometry. In some embodiments, the content of the DP2 fraction is determined by HPLC. In some embodiments, the content of the DP2 fraction is determined by LC-MS / MS or GC-MS.

[0084] In some embodiments, a herein described oligosaccharide preparation has a DP3 fraction content of from about 1% to about 30%, from about 1% to about 25%, from about 1% to about 20%, from about 1% to about 15%, from about 1% to about 10%, from about 5% to about 30%, from about 5% to about 25%, from about 5% to about 20%, from about 5% to about 15%, or from about 5% to about 10% by weight or by relative abundance. In some embodiments, the oligosaccharide preparation has a DP3 fraction content of from about 1% to about 15%, from about 1% to about 10%, from about 5% to about 15%, or from about 5% to about 10% by weight or by relative abundance. In some embodiments, the content of the DP3 fraction is determined by MALDI-MS. In some embodiments, the content of the DP3 fraction is determined by HPLC. In some embodiments, the content of the DP3 fraction is determined by LC-MS / MS or GC-MS.

[0085] In some embodiments, a herein described oligosaccharide preparation has a DP4 fraction content of from about 0.1% to about 20%, from about 0.1% to about 15%, from about 0.1% to about 10%, from about 0.1% to about 5%, from about 1% to about 20%, from about 1% to about 15%, from about 1% to about 10%, or from about 1% to about 5% by weight or by relative abundance. In some embodiments, the oligosaccharide preparation has a DP4 fraction content of from about 1% to about 15%, from about 1% to about 10%, or from about 1% to about 5% by weight or by relative abundance. In some embodiments, a herein described oligosaccharide preparation has a DP5 fraction content of from about 0.1% to about 15%, from about 0.1% to about 10%, from about 0.1% to about 5%, from about 1% to about 15%, from about 1% to about 10%, or from about 1% to about 5% by weight or by relative abundance. In some embodiments, the oligosaccharide preparation has a DP5 fraction content of from about 1% to about 10% or from about 1% to about 5% by weight or by relative abundance. In some embodiments, the content of the DP4 and / or the DP5 fraction is determined by MALDI-MS. In some embodiments, the content of the DP4 and / or the DP5 fraction is determined by HPLC. In some embodiments, the content of the DP4 and / or the DP5 fraction is determined by LC-MS / MS or GC-MS.

[0086] In some embodiments, the ratio of DP2 fraction to DP1 fraction in the oligosaccharide preparation is from about 0.01 to about 0.8, from about 0.02 to about 0.7, from about 0.02 to about 0.6, from about 0.02 to about 0.5, from about 0.02 to about 0.4, from about 0.02 to about 0.3, from about 0.02 to about 0.2, from about 0.1 to about 0.6, from about 0.1 to about 0.5, from about 0.1 to about 0.4, or from about 0.1 to about 0.3 by their weight or relative abundance. In some embodiments, the ratio of DP2 fraction to DP1 fraction in the oligosaccharide preparation is from about 0.02 to about 0.4 by their weight or relative abundance.

[0087] In some embodiments, the ratio of DP3 fraction to DP2 fraction in the oligosaccharide preparation is from about 0.01 to about 0.7, from about 0.01 to about 0.6, from about 0.01 to about 0.5, from about 0.01 to about 0.4, from about 0.01 to about 0.3, or from about 0.01 to about 0.2 by their weight or relative abundance. In some embodiments, the ratio of DP3 fraction to DP2 fraction in the oligosaccharide preparation is from about 0.01 to about 0.3 by their weight or relative abundance.

[0088] In some embodiments, the aggregate content of DP1 and DP2 fractions in the oligosaccharide preparation is less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% by weight or by relative abundance. In some embodiments, the aggregate content of DP1 and DP2 fractions in the oligosaccharide preparation is less than 50%, less than 30%, or less than 10% by weight or by relative abundance.

[0089] In some embodiments, an oligosaccharide preparation described herein has a mean DP value within a range of 2 to 10. In some embodiments, the oligosaccharide preparation has a mean DP value of from about 2 to about 8, from about 2 to about 5, or from about 2 to about 4. In some embodiments, the oligosaccharide preparation has a mean DP value of about 3.5. The mean DP value can be determined by SEC or by elemental analysis. Anhydro-subunit Level

[0090] In some embodiments, a herein described oligosaccharide preparation comprises one or more anhydro-subunits, i.e., the oligosaccharide preparation comprises one or more anhydro- subunit containing oligosaccharides. In some embodiments, each of the n fractions of oligosaccharides in a herein described oligosaccharide preparation independently comprises an anhydro-subunit level. For instance, in some embodiments, the DP1 fraction comprises about 10% of anhydro-subunit containing oligosaccharides by relative abundance, and the DP2 fraction comprises about 15% of anhydro-subunit containing oligosaccharides by relative abundance. For another example, in some embodiments, DP1, DP2, and DP3 fractions each comprises about 5%, about 10%, and about 2% of anhydro-subunit containing oligosaccharides by relative abundance, respectively. In some embodiments, two or more fractions of oligosaccharides comprise similar levels of anhydro-subunit containing oligosaccharides. For example, in some embodiments, the DP1 and DP3 fractions each comprises about 5% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, a herein described oligosaccharide preparation does not comprise any anhydro-subunit, i.e., the oligosaccharide preparation does not comprise any anhydro-subunit containing oligosaccharides.

[0091] In some embodiments, each of the 1 to n fractions in a herein described oligosaccharide preparation independently comprises from about 0.1% to 15% of anhydro-subunit containing oligosaccharides by relative abundance as measured by mass spectrometry, LC-MS / MS or GC- MS. In some embodiments, each of the 1 to n fractions in the oligosaccharide preparation independently comprises from about 0.5% to 15% of anhydro-subunit containing oligosaccharides by relative abundance as measured by mass spectrometry, LC-MS / MS or GC-MS. In some embodiments, LC-MS / MS is used to determine the relative abundance for oligosaccharides in the DP1, DP2, and / or DP3 fractions.

[0092] In some embodiments, the presence, the type of species, and / or level of anhydro-subunits can be determined and / or detected by any suitable analytical methods, such as nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry, HPLC, FFF, A4F, or any combination thereof. In some embodiments, the presence and level of anhydro-subunits containing oligosaccharides are determined and / or detected by MALDI-MS, as illustrated by the -18 g / mol MW offset peaks in FIG. 2. In some embodiments, the presence and the type of species of anhydro-subunits are determined and / or detected by NMR, as illustrated by Example 11, FIG. 3, and FIG. 4. In some embodiments, the presence, the type of species, and / or level of anhydro- subunits or anhydro-subunit containing oligosaccharides is determined and / or detected, at least in part, by mass spectrometry such as MALDI-MS. In some embodiments, the presence, the type of species, and / or level of anhydro-subunits or anhydro-subunit containing oligosaccharides is determined and / or detected, at least in part, by NMR. In some embodiments, the presence, the type of species, and / or level of anhydro-subunits or anhydro-subunit containing oligosaccharides is determined and / or detected, at least in part, by HPLC. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by MALDI-MS. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by LC-MS / MS, as illustrated in FIGs. 21A-21C, 22A-22C, 23A-23C and 24A-24C. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by GC-MS, as illustrated in FIGs. 25A-25B, 26A-26B, 27A-27B and 28A-28B. In some embodiments, GC-MS or LC-MS / MS is used to determine the relative abundance for oligosaccharides in the DP1, DP2, and / or DP3 fractions. In some embodiments, MALDI-MS is used to determine the relative abundance for oligosaccharides in the DP4 fraction or in a higher DP fraction. In some embodiments, the relative abundance of a certain fraction is determined by integrating the area under the peaks of the LC-MS / MS chromatogram that are designated as corresponding to that fraction. In some embodiments, the relative abundance of a certain fraction is determined by integrating the area under the peaks of the GC-MS chromatogram that are designated as corresponding to that fraction.

[0093] In some embodiments, at least one fraction of a herein described oligosaccharide preparation comprises less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, less than 19%, less than 18%, less than 17%, less than 16%, less than 15%, less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, at least one fraction of a herein described oligosaccharide preparation comprises less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, or less than 2% of anhydro-subunit containing oligosaccharides by relative abundance. In other embodiments, at least one fraction of a herein described oligosaccharide preparation comprises greater than 0.5%, greater than 0.8%, greater than 1%, greater than 2%, greater than 3%, greater than 4%, greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, greater than 15%, greater than 16%, greater than 17%, greater than 18%, greater than 19%, greater than 20%, greater than 30%, greater than 40%, greater than 50%, greater than 60%, greater than 70%, or greater than 80% of anhydro-subunit containing oligosaccharides by relative abundance. In other embodiments, at least one fraction of a herein described oligosaccharide preparation comprises greater than 20%, greater than 21%, greater than 22%, greater than 23%, greater than 24%, greater than 25%, greater than 26%, greater than 27%, greater than 28%, greater than 29%, or greater than 30% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, at least one fraction (such as DP1, DP2, and / or DP3) of the oligosaccharide preparation comprises about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 1.5%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, or about 30% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, at least one fraction (such as DP1, DP2, and / or DP3) of the oligosaccharide preparation comprises about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, at least one fraction (such as DP1, DP2, and / or DP3) of the oligosaccharide preparation comprises from about 0.1% to about 90%, from about 0.5% to about 90%, from about 0.5% to about 80%, from about 0.5% to about 70%, from about 0.5% to about 60%, from about 0.5% to about 50%, from about 0.5% to about 40%, from about 0.5% to about 30%, from about 0.5% to about 20%, from about 0.5% to about 10%, from about 0.5% to about 9%, from about 0.5% to about 8%, from about 0.5% to about 7%, from about 0.5% to about 6%, from about 0.5% to about 5%, from about 0.5% to about 4%, from about 0.5% to about 3%, from about 0.5% to about 2%, from about 2% to about 9%, from about 2% to about 8%, from about 2% to about 7%, from about 2% to about 6%, from about 2% to about 5%, from about 2% to about 4%, from about 2% to about 3%, or from about 5% to about 10% of anhydro- subunit containing oligosaccharides by relative abundance. In some embodiments, the relative abundance is measured by mass spectrometry, LC-MS / MS, or GC-MS. In some embodiments, the DP1 and DP2 fractions each independently comprises from about 0.5% to about 15% of anhydro- subunit containing oligosaccharides by relative abundance as measured by mass spectrometry or by LC-MS / MS or GC-MS.

[0094] In some embodiments, each fraction of a herein described oligosaccharide preparation comprises less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, less than 19%, less than 18%, less than 17%, less than 16%, less than 15%, less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, each fraction of a herein described oligosaccharide preparation comprises less than 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, or 2% anhydro-subunit containing oligosaccharides by relative abundance. In other embodiments, each fraction of a herein described oligosaccharide preparation comprises greater than 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, 50%, 60%, 70%, or 80% of anhydro-subunit containing oligosaccharides by relative abundance. In other embodiments, each fraction of a herein described oligosaccharide preparation comprises greater than 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, each fraction of a herein described oligosaccharide preparation comprises about <semantics>0.1%<annotation encoding="application / x-tex">0.1\%< / annotation>< / semantics>, about <semantics>0.2%<annotation encoding="application / x-tex">0.2\%< / annotation>< / semantics>, about <semantics>0.3%<annotation encoding="application / x-tex">0.3\%< / annotation>< / semantics>, about <semantics>0.4%<annotation encoding="application / x-tex">0.4\%< / annotation>< / semantics>, about <semantics>0.5%<annotation encoding="application / x-tex">0.5\%< / annotation>< / semantics>, about <semantics>0.6%<annotation encoding="application / x-tex">0.6\%< / annotation>< / semantics>, about <semantics>0.7%<annotation encoding="application / x-tex">0.7\%< / annotation>< / semantics>, about 0.8%, about 0.9%, about 1%, about 1.5%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, or about 30% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, each fraction of a herein described oligosaccharide preparation comprises about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0,8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, each fraction of a herein described oligosaccharide preparation comprises from about 0.1% to about 90%, from about 0.1% to about 15%, from about 0.5% to about 90%, from about 0.5% to about 80%, from about 0.5% to about 70%, from about 0.5% to about 60%, from about 0.5% to about 50%, from about 0.5% to about 40%, from about 0.5% to about 30%, from about 0.5% to about 20%, from about 0.5% to about 10%, from about 0.5% to about 9%, from about 0.5% to about 8%, from about 0.5% to about 7%, from about 0.5% to about 6%, from about 0.5% to about 5%, from about 0.5% to about 4%, from about 0.5% to about 3%, from about 0.5% to about 2%, from about 2% to about 9%, from about 1% to about 10%, from about 2% to about 8%, from about 2% to about 7%, from about 2% to about 6%, from about 2% to about 5%, from about 2% to about 4%, from about 2% to about 3%, or from about 5% to about 10% of anhydro-subunit containing oligosaccharides by relative abundance.

[0095] In some embodiments, a herein described oligosaccharide preparation comprises less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, less than 19%, less than 18%, less than 17%, less than 16%, less than 15%, less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of anhydro- subunit containing oligosaccharides by relative abundance. In some embodiments, the oligosaccharide preparation comprises less than 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% anhydro-subunit containing oligosaccharides by relative abundance. In other embodiments, the oligosaccharide preparation comprises greater than greater than 0.5%, greater than 0.8%, greater than 1%, greater than 2%, greater than 3%, greater than 4%, greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, greater than 15%, greater than 16%, greater than 17%, greater than 18%, greater than 19%, greater than 20%, greater than 30%, greater than 40%, greater than 50%, greater than 60%, greater than 70%, or greater than 80% of anhydro-subunit containing oligosaccharides by relative abundance. In other embodiments, the oligosaccharide preparation comprises greater than 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the oligosaccharide preparation comprises about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, or about 30% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the oligosaccharide preparation comprises about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 1.5%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the oligosaccharide preparation comprises from about 0.1% to about 90%, from about 0.1% to about 15%, from about 0.5% to about 90%, from about 0.5% to about 80%, from about 0.5% to about 70%, from about 0.5% to about 60%, from about 0.5% to about 50%, from about 0.5% to about 40%, from about 0.5% to about 30%, from about 0.5% to about 20%, from about 0.5% to about 10%, from about 0.5% to about 9%, from about 0.5% to about 8%, from about 0.5% to about 7%, from about 0.5% to about 6%, from about 0.5% to about 5%, from about 0.5% to about 4%, from about 0.5% to about 3%, from about 0.5% to about 2%, from about 2% to about 9%, from about 2% to about 8%, from about 2% to about 7%, from about 2% to about 6%, from about 2% to about 5%, from about 2% to about 4%, from about 2% to about 3%, or from about 5% to about 10% of anhydro-subunit containing oligosaccharides by relative abundance.

[0096] In some embodiments, the DP1 fraction of a herein described oligosaccharide preparation comprises less than 30%, less than 20%, less than 19%, less than 18%, less than 17%, less than 16%, less than 15%, less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the DP1 fraction of a herein described oligosaccharide preparation comprises greater than 0.1%, greater than 0.5%, greater than 0.8%, greater than 1%, greater than 1.5%, greater than 2%, greater than 3%, greater than 4%, greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, or greater than 15% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the DP1 fraction of a herein described oligosaccharide preparation comprises about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the DP1 fraction of a herein described oligosaccharide preparation comprises from about 0.1% to about 15%, from about 0.1% to about 20%, from about 0.5% to about 20%, from 0.5% to about 10%, from about 0.5% to about 15%, from about 1% to about 20%, from about 1% to about 15%, from about 1% to about 10%, from about 2% to about 14%, from about 3% to about 13%, from about 4% to about 12%, from about 5% to about 11%, from about 5% to about 10%, from about 6% to about 9%, or from about 7% to about 8% of anhydro-subunit containing oligosaccharides by relative abundance, or any ranges therebetween. In some embodiments, the DP1 fraction of a herein described oligosaccharide preparation comprises from about 5% to about 10% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by mass spectrometry. In some embodiments, the relative abundance of anhydro- subunit containing oligosaccharides is determined by LC-MS / MS. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by GC-MS.

[0097] In some embodiments, the DP2 fraction of a herein described oligosaccharide preparation comprises less than 30%, less than 20%, less than 19%, less than 18%, less than 17%, less than 16%, less than 15%, less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the DP2 fraction of a herein described oligosaccharide preparation comprises greater than 0.1%, greater than 0.5%, greater than 0.8%, greater than 1%, greater than 1.5%, greater than 2%, greater than 3%, greater than 4%, greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, or greater than 15% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the DP2 fraction of a herein described oligosaccharide preparation comprises about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the DP2 fraction of a herein described oligosaccharide preparation comprises from about 0.1% to about 15%, from about 0.1% to about 20%, from about 0.5% to about 20%, from 0.5% to about 10%, from about 0.5% to about 15%, from about 1% to about 20%, from about 1% to about 15%, from about 1% to about 10%, from about 2% to about 14%, from about 3% to about 13%, from about 4% to about 12%, from about 5% to about 11%, from about 5% to about 10%, from about 6% to about 9%, or from about 7% to about 8% of anhydro-subunit containing oligosaccharides by relative abundance, or any ranges therebetween.. In some embodiments, the DP2 fraction of a herein described oligosaccharide preparation comprises from about 5% to about 10% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by mass spectrometry such as MALDI-MS. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by LC-MS / MS. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by GC-MS.

[0098] In some embodiments, the DP3 fraction of a herein described oligosaccharide preparation comprises less than 30%, less than 20%, less than 19%, less than 18%, less than 17%, less than 16%, less than 15%, less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the DP3 fraction of a herein described oligosaccharide preparation comprises greater than 0.1%, greater than 0.5%, greater than 0.8%, greater than 1%, greater than 1.5%, greater than 2%, greater than 3%, greater than 4%, greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, or greater than 15% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the DP3 fraction of a herein described oligosaccharide preparation comprises about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the DP3 fraction of a herein described oligosaccharide preparation comprises from about 0.1% to about 15%, from about 0.1% to about 20%, from about 0.5% to about 20%, from 0.5% to about 10%, from about 0.5% to about 15%, from about 1% to about 20%, from about 1% to about 15%, from about 1% to about 10%, from about 2% to about 14%, from about 3% to about 13%, from about 4% to about 12%, from about 5% to about 11%, from about 5% to about 10%, from about 6% to about 9%, or from about 7% to about 8% of anhydro-subunit containing oligosaccharides by relative abundance, or any ranges therebetween. In some embodiments, the DP3 fraction of a herein described oligosaccharide preparation comprises from about 5% to about 10% of anhydro-subunit containing oligosaccharides by relative abundance. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by mass spectrometry such as MALDI-MS. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by LC-MS / MS. In some embodiments, the relative abundance of anhydro-subunit containing oligosaccharides is determined by GC-MS.

[0099] In some embodiments, an anhydro-subunit containing oligosaccharide comprises one or more anhydro-subunits. For instance, a DP1 anhydro-subunit containing oligosaccharide comprises one anhydro-subunit. In some embodiments, a DPn anhydro-subunit containing oligosaccharide can comprise from 1 to n anhydro-subunits. For example, in some embodiments, a DP2 anhydro-subunit containing oligosaccharide comprises one or two anhydro-subunits. In some embodiments, each oligosaccharide in the oligosaccharide preparation independently comprises zero, one, or two anhydro-subunits. In some embodiments, more than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, or 30% of the anhydro-subunit containing oligosaccharides have only one anhydro-subunit. In some embodiments, more than 99%, 95%, 90%, 85%, or 80% of the anhydro-subunit containing oligosaccharides have only one anhydro-subunit.

[00100] In some embodiments, one or more oligosaccharides in the oligosaccharide preparation or in each fraction of the oligosaccharide preparation comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 anhydro-subunits each linked via a glycosidic bond, wherein the glycosidic bond linking each anhydro-subunit are independently chosen. In some embodiments, one or more oligosaccharides in the oligosaccharide preparation or in each fraction of the oligosaccharide preparation comprise 1, 2, or 3 anhydro-subunits each linked via a glycosidic bond, wherein the glycosidic bond linking each anhydro-subunit are independently chosen. In some embodiments, greater than 50%, 60%, 70%, 80%, 90%, or 99% of oligosaccharides in the oligosaccharide preparation or in each fraction comprise 1, 2, or 3 anhydro-subunits each linked via a glycosidic bond, wherein the glycosidic bond linking each anhydro-subunit are independently chosen. In some embodiments, one or more oligosaccharides in the oligosaccharide preparation or in each fraction comprise 1 anhydro- subunit linked via a glycosidic bond. In some embodiments, greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, or greater than 99% of oligosaccharides in the oligosaccharide preparation or in each fraction comprise 1 anhydro-subunit linked via a glycosidic bond. Anhydro-subunit Species

[00101] In some embodiments, the oligosaccharide preparation comprises different species of anhydro-subunits. In some embodiments, exemplary anhydro-subunit containing oligosaccharides are illustrated in FIG. 5, FIG. 18, and FIG. 19. In some embodiments, the oligosaccharide preparation comprises one or more anhydro-subunits that are products of thermal dehydration of monosaccharides, i.e., anhydro-monosaccharide subunits. In some embodiments, the oligosaccharide preparation comprises one or more anhydro-subunits that are products of reversible thermal dehydration of monosaccharides.

[00102] It is to be understood that an anhydro-monosaccharide (or an anhydro-monosaccharide subunit) refers to one or more species of the thermal dehydration products of the monosaccharide. For example, in some embodiments, an anhydro-glucose refers to 1,6-anhydro-β-D- glucopyranose (levoglucosan) or 1,6-anhydro-β-D-glucofuranose. In some embodiments, a plurality of anhydro-glucose refer to a plurality of 1,6-anhydro-β-D-glucopyranose (levoglucosan), a plurality of 1,6-anhydro-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-D-glucofuranose, a plurality of other thermal dehydration products of glucose, or any combination thereof. Similarly, in some embodiments, a plurality of anhydro-galactose refers to a plurality of any thermal dehydration products of galactose, or any combination thereof.

[00103] In some embodiments, an oligosaccharide preparation as described herein comprises one or more anhydro-glucose, anhydro-galactose, anhydro-mannose, anhydro-allose, anhydro-altrose, anhydro-gulose, anhydro-indose, anhydro-talose, anhydro-fructose, anhydro-ribose, anhydro- arabinose, anhydro-rhamnose, anhydro-lyxose, anhydro-xylose, or any combination of these subunits. In some embodiments, the oligosaccharide preparation comprises one or more anhydro- glucose, anhydro-galactose, anhydro-mannose, or anhydro-fructose subunits. In some embodiments, an oligosaccharide preparation as described herein comprises one or more of: 1,6- anhydro-3-O-β-D-glucopyranosyl-β-D-glucopyranose, 1,6-anhydro-3-O-α-D-glucopyranosyl-β- D-glucopyranose, 1,6-anhydro-2-O-β-D-glucopyranosyl-β-D-glucopyranose, 1,6-anhydro-2-O- α-D-glucopyranosyl-β-D-glucopyranose, 1,6-anhydro-β-D-cellobiose (cellobiosan), 1,6-anhydro- β-D-cellotriose (cellotriosan), 1,6-anhydro-β-D-cellotetraose (cellotetraosan), 1,6-anhydro-β-D- cellopentaose (cellopentaosan), and 1,6-anhydro-β-D-maltose (maltosan).

[00104] In some embodiments, the oligosaccharide preparation comprises one or more 1,6- anhydro-β-D-glucofuranose subunits. In some embodiments, the oligosaccharide preparation comprises one or more 1,6-anhydro-β-D-glucopyranose (levoglucosan) subunits. For example, FIG. 18 illustrates two DP1 anhydro-subunit containing oligosaccharides (levoglucosan and 1,6- anhydro-β-D-glucofuranose) and a DP2 anhydro-subunit containing oligosaccharide (anhydro- cellobiose).

[00105] The presence and the level of a species of anhydro-subunit can vary based on the feed sugars used to manufacture the oligosaccharide. For instance, in some embodiments, gluco- oligosaccharides comprise anhydro-glucose subunits, galacto-oligosaccharides comprise anhydro-galactose subunits, and gluco-galacto-oligosaccharides comprise anhydro-glucose and anhydro-galactose subunits.

[00106] In some embodiments, the oligosaccharide preparation comprises both 1,6-anhydro-β-D- glucofuranose and 1,6-anhydro-β-D-glucopyranose anhydro-subunits. In some embodiments, at least 0.1%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 99% of anhydro- subunits are selected from a group consisting of 1,6-anhydro-β-D-glucofuranose and 1,6-anhydro- β-D-glucopyranose. In some embodiments, at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of anhydro-subunits are 1,6-anhydro-β-D-glucofuranose. In some embodiments, at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, or 60% of anhydro-subunits are 1,6- anhydro-β-D-glucopyranose.

[00107] In some embodiments, the ratio of 1,6-anhydro-β-D-glucofuranose to 1,6-anhydro-β-D- glucopyranose is from about 10:1 to 1:10, 9:1 to 1:10, 8:1 to 1:10, 7:1 to 1:10, 6:1 to 1:10, 5:1 to 1:10, 4:1 to 1:10, 3:1 to 1:10, 2:1 to 1:10, 10:1 to 1:9, 10:1 to 1:8, 10:1 to 1:7, 10:1 to 1:6, 10:1 to 1:5, 10:1 to 1:4, 10:1 to 1:3, 10:1 to 1:2, or 1:1 to 3:1 in the preparation. In some embodiments, the ratio of 1,6-anhydro-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-D-glucofuranose to 1,6-anhydro-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-D-glucopyranose is about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:8, 1:9, or 1:10 in the preparation. In some embodiments, the ratio of 1,6-anhydro-β-D-glucofuranose to 1,6-anhydro- <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-D-glucopyranose is about 2:1 in the preparation.

[00108] In some embodiments, the ratio of 1,6-anhydro-β-D-glucofuranose to 1,6-anhydro-β-D- glucopyranose is about from 10:1 to 1:10, 9:1 to 1:10, 8:1 to 1:10, 7:1 to 1:10, 6:1 to 1:10, 5:1 to 1:10, 4:1 to 1:10, 3:1 to 1:10, 2:1 to 1:10, 10:1 to 1:9, 10:1 to 1:8, 10:1 to 1:7, 10:1 to 1:6, 10:1 to 1:5, 10:1 to 1:4, 10:1 to 1:3, 10:1 to 1:2, or 1:1 to 3:1 in each fraction. In some embodiments, the ratio of 1,6-anhydro-β-D-glucofuranose to 1,6-anhydro-β-D-glucopyranose is about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:8, 1:9, or 1:10 in each fraction. In some embodiments, the ratio of 1,6-anhydro-β-D-glucofuranose to 1,6-anhydro-β-D- glucopyranose is about 2:1 in each fraction.

[00109] In some embodiments, the ratio of 1,6-anhydro-β-D-glucofuranose to 1,6-anhydro-β-D- glucopyranose is about from 10:1 to 1:10, 9:1 to 1:10, 8:1 to 1:10, 7:1 to 1:10, 6:1 to 1:10, 5:1 to 1:10, 4:1 to 1:10, 3:1 to 1:10, 2:1 to 1:10, 10:1 to 1:9, 10:1 to 1:8, 10:1 to 1:7, 10:1 to 1:6, 10:1 to 1:5, 10:1 to 1:4, 10:1 to 1:3, 10:1 to 1:2, or 1:1 to 3:1 in at least one fraction. In some embodiments, the ratio of 1,6-anhydro-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-D-glucofuranose to 1,6-anhydro-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-D-glucopyranose is about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:8, 1:9, or 1:10 in at least one fraction. In some embodiments, the ratio of 1,6-anhydro-β-D-glucofuranose to 1,6-anhydro-β-D- glucopyranose is about 2:1 in at least one fraction.

[00110] In some embodiments, a herein described oligosaccharide preparation comprises anhydro-subunit containing DP2 oligosaccharides. In some embodiments, the oligosaccharide preparation comprises anhydro-lactose, anhydro-sucrose, anhydro-cellobiose, or a combination thereof. In some embodiment, the oligosaccharide preparation comprises from about 2 to 20, 2 to 15, 5 to 20, 5 to 15, or 5 to 10 species of DP2 anhydro-subunit containing oligosaccharides. In some embodiments, an oligosaccharide preparation described herein does not comprise cellobiosan or does not comprise a detectable level of cellobiosan.

[00111] In some embodiments, a herein described oligosaccharide preparation comprises one or more anhydro-subunits that are sugar caramelization products. In some embodiments, the oligosaccharide preparation comprises one or more anhydro-subunits are sugar caramelization products selected from the group consisting of: methanol; ethanol; furan; methyl glyoxal; 2- methyl furan; vinyl acetate; glycolaldehyde; acetic acid; acetol; furfural; 2-furanmethanol; 3- furanmethanol; 2-hydroxy cyclopent-2-en-1-one; 5-methyl furfural; 2(5H)-furanone; 2 methyl cyclopentenolone; levoglucosenone; cyclic hydroxyl lactone; 1,4,3,6-dianhydro-α-D- glucopyranose; dianhydro glucopyranose; and 5-hydroxy methyl furfural (5-hmf).

[00112] In some embodiments, in the oligosaccharide preparation or in at least one of the DP fractions, the anhydro-subunits that are caramelization products are less abundant than the anhydro-subunits that are products of reversible thermal dehydration of a monosaccharide. In some embodiments, in the oligosaccharide preparation or in at least one of the fractions, the anhydro-subunits that are caramelization products are more abundant than the anhydro-subunits that are products of reversible thermal dehydration of a monosaccharide. In some embodiments, in the oligosaccharide preparation or in at least one of the fractions, anhydro-subunits that are caramelization products and anhydro-subunits that are products of reversible thermal dehydration of a monosaccharide have similar abundance.

[00113] In some embodiments, from about 0.01% to about 50%, from about 0.01% to about 40%, from about 0.01% to about 30%, from about 0.01% to about 20%, from about 0.01% to about 10%, from about 0.01% to about 5%, from about 0.01% to about 4%, from about 0.01% to about 3%, from about 0.01% to about 2%, from about 0.01% to about 1%, from about 0.01% to about 0.5%, from about 0.1% to about 50%, from about 0.1% to about 40%, from about 0.1% to about 30%, from about 0.1% to about 20%, from about 0.1% to about 10%, from about 0.1% to about 5%, from about 0.1% to about 4%, from about 0.1% to about 3%, from about 0.1% to about 2%, from about 0.1% to about 1%, or from about 0.1% to about 0.5% of the anhydro-subunits in a herein described oligosaccharide preparation are caramelization products. In some embodiments, from about 0.1% to about 5%, from about 0.1% to about 2%, or from about 0.1% to about 1% of the anhydro-subunits in the oligosaccharide preparation are caramelization products. In some embodiments, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 15%, less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of the anhydro-subunits in the oligosaccharide preparation are caramelization products.

[00114] In some embodiments, from about 0.01% to about 50%, from about 0.01% to about 40%, from about 0.01% to about 30%, from about 0.01% to about 20%, from about 0.01% to about 10%, from about 0.01% to about 5%, from about 0.01% to about 4%, from about 0.01% to about 3%, from about 0.01% to about 2%, from about 0.01% to about 1%, from about 0.01% to about 0.5%, from about 0.1% to about 50%, from about 0.1% to about 40%, from about 0.1% to about 30%, from about 0.1% to about 20%, from about 0.1% to about 10%, from about 0.1% to about 5%, from about 0.1% to about 4%, from about 0.1% to about 3%, from about 0.1% to about 2%, from about 0.1% to about 1%, or from about 0.1% to about 0.5% of the anhydro-subunits in at least one fraction (e.g., DP1, DP2 and / or DP3) of a herein described preparation are caramelization products. In some embodiments, from about 0.1% to about 5%, from about 0.1% to about 2 %, or from about 0.1% to about 1% of the anhydro-subunits in at least one fraction (e.g., DP1, DP2) and / or DP3) of the preparation are caramelization products. In some embodiments, less than 50%, 40%, 30%, 25%, 20%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the anhydro-subunits in at least one fraction of the preparation are caramelization products. In some embodiments, less than 20%, less than 15%, less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of the anhydro-subunits in the DP1, DP2, and / or DP3 fractions of a herein described oligosaccharide preparation are caramelization products.

[00115] In some embodiments, from about 0.01% to about 50%, from about 0.01% to about 40%, from about 0.01% to about 30%, from about 0.01% to about 20%, from about 0.01% to about 10%, from about 0.01% to about 5%, from about 0.01% to about 4%, from about 0.01% to about 3%, from about 0.01% to about 2%, from about 0.01% to about 1%, from about 0.01% to about 0.5%, from about 0.1% to about 50%, from about 0.1% to about 40%, from about 0.1% to about 30%, from about 0.1% to about 20%, from about 0.1% to about 10%, from about 0.1% to about 5%, from about 0.1% to about 4%, from about 0.1% to about 3%, from about 0.1% to about 2%, from about 0.1% to about 1%, or from about 0.1% to about 0.5% of the anhydro-subunits in each fraction of a herein described oligosaccharide preparation are caramelization products. In some embodiments, from about 0.1% to about 5%, from about 0.1% to about 2%, or from about 0.1% to about 1% of the anhydro-subunits in each fraction of the preparation are caramelization products. In some embodiments, less than 50%, less than 40%, less than 30%, less than 20%, less than 25%, less than 20%, less than 15%, less than 14%, less than 13%, less than 12%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of the anhydro-subunits in each fraction of the preparation are caramelization products.

[00116] In some embodiments, each of the oligosaccharides in a herein described oligosaccharide preparation independently and optionally comprises an anhydro-subunit. In some embodiments, two or more independent oligosaccharides comprise the same or different anhydro-subunits. In some embodiments, two or more independent oligosaccharides comprise different anhydro- subunits. For example, in some embodiments, the oligosaccharide preparation comprise a DP1 anhydro-subunit containing oligosaccharide that comprises a 1,6-anhydro-β-D-glucopyranose and a DP2 anhydro-subunit containing oligosaccharide that comprises a 1,6-anhydro-β-D- glucofuranose subunit. In some embodiments, one or more oligosaccharides in the oligosaccharide preparation comprise two or more the same or different anhydro-subunits.

[00117] In some embodiments, in any fraction of the oligosaccharide preparation that has a degree of polymerization equal or greater than 2 (i.e., DP2 to DPn fractions), an anhydro-subunit can be linked to one or more regular or anhydro-subunits. In some embodiments, in the DP2 to DPn fractions, at least one anhydro-subunit is linked to one, two, or three other regular or anhydro- subunits. In some embodiments, in the DP2 to DPn fractions, at least one anhydro-subunit is linked to one or two regular subunits. In some embodiments, in the DP2 to DPn fractions, at least one anhydro-subunit is linked to one regular subunit. In some embodiments, in any of the DP2 to DPn fractions, more than 99%, 90%, 80%, 70%, 60%, 50%, 40%, or 30% of anhydro-subunits are linked to one regular subunit. In some embodiments, in each of the DP2 to DPn fraction, more than 99%, 90%, 80%, 70%, 60%, 50%, 40%, or 30% of anhydro-subunits are linked to one regular subunit.

[00118] In some embodiments, in any fraction of the oligosaccharide preparation that has a degree of polymerization equal or greater than 2 (i.e., DP2 to DPn fractions), an anhydro-subunit can be located at a chain-end of an oligosaccharide. In some embodiments, in any fraction of the oligosaccharide preparation that has a degree of polymerization equal or greater than 3 (i.e., DP3) to DPn fractions), an anhydro-subunit can be located at a position that is not a chain-end of an oligosaccharide. In some embodiments, greater than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, or 30% of the anhydro-subunits in the DP2 to DPn fractions are located at the chain-end of the oligosaccharides. In some embodiments, greater than 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the anhydro-subunit containing oligosaccharides comprise a chain-end anhydro-subunit. In some embodiments, greater than 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the anhydro-subunit containing oligosaccharides comprise a chain-end anhydro-subunit.

[00119] In some embodiments, greater than 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the anhydro-subunit containing oligosaccharides comprise a chain-end anhydro-subunit. In some embodiments, greater than 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the anhydro-subunit containing oligosaccharides comprise a chain-end anhydro- subunit. Glycosidic Linkages

[00120] In some embodiments, a herein described oligosaccharide preparation used in the methods described herein comprises a variety of glycosidic linkages. The type and distribution of the glycosidic linkages can depend on the source and manufacturing method of the oligosaccharide preparation. In some embodiments, the type and distribution of various glycosidic linkages can be determined and / or detected by any suitable methods known in the art such as NMR. For example, in some embodiments, the glycosidic linkages are determined and / or detected by 1H NMR, 13C NMR, 2D NMR such as 2D JRES, HSQC, HMBC, DOSY, COSY, ECOSY, TOCSY, NOESY, or ROESY, or any combination thereof. In some embodiments, the glycosidic linkages are determined and / or detected, at least in part, by proton NMR. In some embodiments, the glycosidic linkages are determined and / or detected, at least in part, by 13C NMR. In some embodiments, the glycosidic linkages are determined and / or detected, at least in part, by 2D 1H, 13C- HSQC NMR.

[00121] In some embodiments, a herein described oligosaccharide preparation comprises one or more <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, or any combination thereof.

[00122] In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of about from 0 to about 60 mol%, from about 5% to about 55 mol%, from about 5% to about 50 mol%, from about 5% to about 45 mol%, from about 5% to about 40 mol%, from about 5% to about 35 mol%, from about 5% to about 30 mol%, from about 5% to about 25 mol%, from about 10% to about 60 mol%, from about 10% to about 55 mol%, from about 10% to about 50 mol%, from about 10% to about 45 mol%, from about 10% to about 40 mol%, from about 10% to about 35 mol%, from about 15% to about 60 mol%, from about 15% to about 55 mol%, from about 15% to about 50 mol%, from about 15% to about 45 mol%, from about 15% to about 40 mol%, from about 15% to about 35 mol%, from about 20% to about 60 mol%, from about 20% to about 55 mol%, from about 20% to about 50 mol%, from about 20% to about 45 mol%, from about 20% to about 40 mol%, from about 20% to about 35 mol%, from about 25% to about 60 mol%, from about 25% to about 55 mol%, from about 25% to about 50 mol%, from about 25% to about 45 mol%, from about 25% to about 40 mol%, or from about 25% to about 35 mol% of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>- (1,6) glycosidic linkages.

[00123] In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of from about 0 to about 50 mol%, from about 0 to about 40 mol%, from about 0 to about 35 mol%, from about 0 to about 30 mol%, from about 0 to about 25 mol%, from about 0 to about 20 mol%, from about 5% to about 40 mol%, from about 5% to about 35 mol%, from about 5% to about 30 mol%, from about 5% to about 25 mol%, from about 5% to about 20 mol%, from about 10% to about 40 mol%, from about 10% to about 35 mol%, from about 10% to about 20 mol%, from about 15% to about 40 mol%, from about 15% to about 35 mol%, from about 15% to about 30 mol%, from about 15% to about 25 mol%, or from about 15% to about 20 mol% of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>- (1,3) glycosidic linkages.

[00124] In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of from about 0 to about 40 mol%, from about 0 to about 35 mol%, from about 0 to about 30 mol%, from about 0 to about 25 mol%, from about 0 to about 20 mol%, from about 0 to about 15 mol%, from about 0 to about 10 mol%, from about 2% to about 30 mol%, from about 2% to about 25 mol%, from about 2% to about 20 mol%, from about 2% to about 15 mol%, from about 2% to about 10 mol%, from about 3% to about 30 mol%, from about 3% to about 25 mol%, from about 3% to about 20 mol%, from about 3% to about 15 mol%, from about 3% to about 10 mol%, from about 5% to about 30 mol%, from about 5% to about 25 mol%, from about 5% to about 20 mol\%, from about 5\% to about 15 mol\%, or from about 5\% to about 10 mol\% of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages.

[00125] In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of from about 0 to about 40 mol%, from about 0 to about 30 mol%, from about 0 to about 25 mol%, from about 0 to about 20 mol%, from about 0 to about 15 mol%, from about 0 to about 10 mol\%, or from about 0 to about 5 mol\% of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkages. In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of less than 40 mol%, less than 30 mol%, less than 20 mol%, less than 15 mol%, less than 10 mol%, less than 9 mol%, less than 8 mol%, less than 7 mol%, less than 6 mol%, less than 5 mol%, less than 4 mol%, less than 3 mol%, or less than 2 mol% of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkages.

[00126] In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of from about 0 to about 40 mol%, from about 0 to about 35 mol%, from about 0 to about 30 mol%, from about 0 to about 25 mol%, from about 0 to about 20 mol%, from about 0 to about 15 mol%, from about 0 to about 10 mol%, from about 2% to about 30 mol%, from about 2% to about 25 mol%, from about 2% to about 20 mol%, from about 2% to about 15 mol%, from about 2% to about 10 mol%, from about 5% to about 30 mol%, from about 5% to about 25 mol%, from about 5% to about 20 mol%, from about 5% to about 15 mol%, from about 5% to about 10 mol%, from about 8% to about 30 mol%, from about 8% to about 25 mol%, from about 8% to about 20 mol%, from about 8% to about 15 mol%, or from about 10% to about 15 mol% of β- (1,6) glycosidic linkages.

[00127] In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of from about 0 to about 40 mol%, from about 0 to about 35 mol%, from about 0 to about 30 mol%, from about 0 to about 25 mol%, from about 0 to about 20 mol%, from about 0 to about 15 mol%, from about 0 to about 10 mol%, from about 2% to about 30 mol%, from about 2% to about 25 mol%, from about 2% to about 20 mol%, from about 2% to about 15 mol%, from about 2% to about 10 mol%, from about 3% to about 30 mol%, from about 3% to about 25 mol%, from about 3% to about 20 mol%, from about 3% to about 15 mol%, from about 3% to about 10 mol%, from about 5% to about 30 mol%, from about 5% to about 25 mol%, from about 5% to about 20 mol%, from about 5% to about 15 mol%, or from about 5% to about 10 mol% of <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages.

[00128] In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of from about 0 to about 40 mol%, from about 0 to about 30 mol%, from about 0 to about 25 mol%, from about 0 to about 20 mol%, from about 0 to about 15 mol%, from about 0 to about 10 mol%, from about 0 to about 5 mol%, from about 1% to about 20 mol%, from about 1% to about 15 mol%, from about 1% to about 10 mol%, from about 1% to about 5 mol%, from about 2% to about 20 mol%, from about 2% to about 15 mol%, from about 2% to about 10 mol%, or from about 2% to about 5 mol% of <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages. In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of less than 40 mol%, less than 30 mol%, less than 20 mol%, less than 15 mol%, less than 10 mol%, less than 9 mol%, less than 8 mol%, less than 7 mol%, less than 6 mol%, less than 5 mol%, less than 4 mol%, less than 3 mol%, or less than 2 mol% of <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages.

[00129] In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of from about 0 to about 40 mol%, from about 0 to about 30 mol%, from about 0 to about 25 mol%, from about 0 to about 20 mol%, from about 0 to about 15 mol%, from about 0 to about 10 mol%, from about 0 to about 5 mol%, from about 1% to about 20 mol%, from about 1% to about 15 mol%, from about 1% to about 10 mol%, from about 1% to about 5 mol%, from about 2% to about 20 mol%, from about 2% to about 15 mol%, from about 2% to about 10 mol%, or from about 2% to about 5 mol% of <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages. In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution of less than 40 mol%, less than 30 mol%, less than 20 mol%, less than 15 mol%, less than 10 mol%, less than 9 mol%, less than 8 mol%, less than 7 mol%, less than 6 mol%, less than 5 mol%, less than 4 mol%, less than 3 mol%, or less than 2 mol% of <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages.

[00130] In some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution that is different from a glycosidic bond type distribution of non-synthetic oligosaccharide preparations. For example, in some embodiments, the oligosaccharide preparations have a glycosidic bond type distribution that is different from that of the base nutritional compositions. In some embodiments, the base nutritional compositions comprise a natural carbohydrate source, such as starch and plant fibers. Some of the natural carbohydrate sources have a high percentage of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4), <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6), and / or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages. Accordingly, in some embodiments, the oligosaccharide preparations have a lower percentage of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkages than the base nutritional composition. In some embodiments, the oligosaccharide preparations have a lower percentage of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages than the base nutritional composition. In other embodiments, the oligosaccharide preparations have a higher percentage of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages than the base nutritional composition. In some embodiments, the oligosaccharide preparations have a lower percentage of <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages than the base nutritional composition. In some embodiments, the oligosaccharide preparation comprises glycosidic linkages that are not readily digestible or hydrolysable by enzymes.

[00131] Specifically, in some embodiments, the <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2), <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3), <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4), <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6), <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2), <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3), <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4), and / or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages in the glycosidic bond type distribution of a herein described oligosaccharide preparations is at least 50 mol%, at least 40 mol%, at least 30 mol%, at least 20 mol%, at least 15 mol%, at least 10 mol%, at least 5 mol%, at least 2 mol%, or at least 1 mol% lower than that of the base nutritional composition. In some embodiments, the <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2), <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>- <semantics>(1,3)<annotation encoding="application / x-tex">(1,3)< / annotation>< / semantics>, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-<semantics>(1,4)<annotation encoding="application / x-tex">(1,4)< / annotation>< / semantics>, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-<semantics>(1,6)<annotation encoding="application / x-tex">(1,6)< / annotation>< / semantics>, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-<semantics>(1,2)<annotation encoding="application / x-tex">(1,2)< / annotation>< / semantics>, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-<semantics>(1,3)<annotation encoding="application / x-tex">(1,3)< / annotation>< / semantics>, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-<semantics>(1,4)<annotation encoding="application / x-tex">(1,4)< / annotation>< / semantics>, and / or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-<semantics>(1,6)<annotation encoding="application / x-tex">(1,6)< / annotation>< / semantics> glycosidic linkages in the glycosidic bond type distribution of the oligosaccharide preparations is at least 50 mol%, at least 40 mol%, at least 30 mol%, at least 20 mol%, at least 15 mol%, at least 10 mol%, at least 5 mol%, at least 2 mol%, or at least 1 mol% higher than that of the base nutritional composition.

[00132] It should be understood by one of skill in the art that certain types of glycosidic linkages may not be applicable to oligosaccharides comprising certain type of monosaccharides. For example, in some embodiments, the oligosaccharide preparation comprises <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages and <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages. In other embodiments, the oligosaccharide preparation comprises <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages and <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages. In some embodiments, the oligosaccharide preparation comprises <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, and <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages. In some embodiments, the oligosaccharide preparation comprises <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, and <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages. Molecular Weight

[00133] The molecular weight and molecular weight distribution of the herein described oligosaccharide preparations can be determined by any suitable analytical means and instrumentation, such as end group method, osmotic pressure (osmometry), ultracentrifugation, viscosity measurements, light scattering method, SEC, SEC-MALLS, FFF, A4F, HPLC, and mass spectrometry. In some embodiments, the molecular weight and molecular weight distribution are determined by mass spectrometry, such as MALDI-MS, LC-MS, or GC-MS. In some embodiments, the molecular weight and molecular weight distribution are determined by size exclusion chromatography (SEC), such as gel permeation chromatography (GPC). In other embodiments, the molecular weight and molecular weight distribution are determined by HPLC. In some embodiments, the molecular weight and molecular weight distribution are determined by MALDI-MS.

[00134] In some embodiments, a herein described oligosaccharide preparation has a weight average molecular weight of from about 100 to about 10000 g / mol, from about 200 to about 8000 g / mol, from about 300 to about 5000 g / mol, from about 500 to about 5000 g / mol, from about 700 to about 5000 g / mol, from about 900 to about 5000 g / mol, from about 1100 to about 5000 g / mol, from about 1300 to about 5000 g / mol, from about 1500 to about 5000 g / mol, from about 1700 to about 5000 g / mol, from about 300 to about 4500 g / mol, from about 500 to about 4500 g / mol, from about 700 to about 4500 g / mol, from about 900 to about 4500 g / mol, from about 1100 to about 4500 g / mol, from about 1300 to about 4500 g / mol, from about 1500 to about 4500 g / mol, from about 1700 to about 4500 g / mol, from about 1900 to about 4500 g / mol, from about 300 to about 4000 g / mol, from about 500 to about 4000 g / mol, from about 700 to about 4000 g / mol, from about 900 to about 4000 g / mol, from about 1100 to about 4000 g / mol, from about 1300 to about 4000 g / mol, from about 1500 to about 4000 g / mol, from about 1700 to about 4000 g / mol, from about 1900 to about 4000 g / mol, from about 300 to about 3000 g / mol, from about 500 to about 3000 g / mol, from about 700 to about 3000 g / mol, from about 900 to about 3000 g / mol, from about 1100 to about 3000 g / mol, from about 1300 to about 3000 g / mol, from about 1500 to about 3000 g / mol, from about 1700 to about 3000 g / mol, from about 1900 to about 3000 g / mol, from about 2100 to about 3000 g / mol, from about 300 to about 2500 g / mol, from about 500 to about 2500 g / mol, from about 700 to about 2500 g / mol, from about 900 to about 2500 g / mol, from about 1100 to about 2500 g / mol, from about 1300 to about 2500 g / mol, from about 1500 to about 2500 g / mol, from about 1700 to about 2500 g / mol, from about 1900 to about 2500 g / mol, from about 2100 to about 2500 g / mol, from about 300 to about 1500 g / mol, from about 500 to about 1500 g / mol, from about 700 to about 1500 g / mol, from about 900 to about 1500 g / mol, from about 1100 to about 1500 g / mol, from about 1300 to about 1500 g / mol, from about 2000 to about 2800 g / mol, from about 2100 to about 2700 g / mol, from about 2200 to about 2600 g / mol, from about 2300 to about 2500 g / mol, or from about 2320 to about 2420 g / mol. In some embodiments, the weight average molecular weight of the oligosaccharide preparation is from about 2000 to about 2800 g / mol, from about 2100 to about 2700 g / mol, from about 2200 to about 2600 g / mol, from about 2300 to about 2500 g / mol, or from about 2320 to about 2420 g / mol. In some embodiments, the oligosaccharide preparation has a weight average molecular weight in a range from at least 500 g / mol, 750 g / mol, 1000 g / mol, or 1500 g / mol to at most 1750 g / mol, 2000 g / mol, 2250 g / mol, 2500 g / mol, or 3000 g / mol. In some embodiments, the weight average molecular weight of a herein described oligosaccharide preparation is determined by HPLC according to Example 9.

[00135] In some embodiments, a herein described oligosaccharide preparation has a number average molecular weight of from about 100 to about 10000 g / mol, from about 200 to about 8000 g / mol, from about 300 to about 5000 g / mol, from about 500 to about 5000 g / mol, from about 700 to about 5000 g / mol, from about 900 to about 5000 g / mol, from about 1100 to about 5000 g / mol, from about 1300 to about 5000 g / mol, from about 1500 to about 5000 g / mol, from about 1700 to about 5000 g / mol, from about 300 to about 4500 g / mol, from about 500 to about 4500 g / mol, from about 700 to about 4500 g / mol, from about 900 to about 4500 g / mol, from about 1100 to about 4500 g / mol, from about 1300 to about 4500 g / mol, from about 1500 to about 4500 g / mol, from about 1700 to about 4500 g / mol, from about 1900 to about 4500 g / mol, from about 300 to about 4000 g / mol, from about 500 to about 4000 g / mol, from about 700 to about 4000 g / mol, from about 900 to about 4000 g / mol, from about 1100 to about 4000 g / mol, from about 1300 to about 4000 g / mol, from about 1500 to about 4000 g / mol, from about 1700 to about 4000 g / mol, from about 1900 to about 4000 g / mol, from about 300 to about 3000 g / mol, from about 500 to about 3000 g / mol, from about 700 to about 3000 g / mol, from about 900 to about 3000 g / mol, from about 1100 to about 3000 g / mol, from about 1300 to about 3000 g / mol, from about 1500 to about 3000 g / mol, from about 1700 to about 3000 g / mol, from about 1900 to about 3000 g / mol, from about 2100 to about 3000 g / mol, from about 300 to about 2500 g / mol, from about 500 to about 2500 g / mol, from about 700 to about 2500 g / mol, from about 900 to about 2500 g / mol, from about 1100 to about 2500 g / mol, from about 1300 to about 2500 g / mol, from about 1500 to about 2500 g / mol, from about 1700 to about 2500 g / mol, from about 1900 to about 2500 g / mol, from about 2100 to about 2500 g / mol, from about 300 to about 2000 g / mol, from about 500 to about 300 to 2000 g / mol, from about 700 to about 2000 g / mol, from about 900 to about 2000 g / mol, from about 1100 to about 2000 g / mol, from about 300 to about 1500 g / mol, from about 500 to about 1500 g / mol, from about 700 to about 1500 g / mol, from about 900 to about 1500 g / mol, from about 1100 to about 1500 g / mol, from about 1300 to about 1500 g / mol, from about 1000 to about 2000 g / mol, from about 1100 to about 1900 g / mol, from about 1200 to about 1800 g / mol, from about 1300 to about 1700 g / mol, from about 1400 to about 1600 g / mol, or from about 1450 to about 1550 g / mol. In some embodiments, the number average molecular weight of the oligosaccharide preparation is from about 1000 to about 2000 g / mol, from about 1100 to about 1900 g / mol, from about 1200 to about 1800 g / mol, from about 1300 to about 1700 g / mol, 1400 to 1600 g / mol, or 1450-1550 g / mol. In some embodiments, the oligosaccharide preparation has a number average molecular weight in a range from at least 500 g / mol, 750 g / mol, 1000 g / mol, or 1500 g / mol to at most 1750 g / mol, 2000 g / mol, 2250 g / mol, 2500 g / mol, or 3000 g / mol. In some embodiments, the number average molecular weight of a herein described oligosaccharide preparation is determined by HPLC according to Example 9. Types of Oligosaccharides

[00136] In some embodiments, a herein described oligosaccharide preparations comprises one or more species of monosaccharide subunits. In some embodiments, the oligosaccharide preparation can comprise oligosaccharides with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more different species of monosaccharides subunits.

[00137] In some embodiments, a herein described oligosaccharide preparation comprises one or more monosaccharide subunits selected from a group consisting of: triose, tetrose, pentose, hexose, heptose, and any combination thereof, wherein each of the said triose, tetrose, pentose, hexose, or heptose subunit is independently and optionally functionalized and / or replaced with one of its corresponding anhydro-subunits. In some embodiments, the corresponding anhydro- subunit is a product of reversible thermal dehydration of the monosaccharide subunit. In some embodiments, the corresponding anhydro-subunit is a caramelization product of the monosaccharide subunit.

[00138] In some embodiments, a herein described oligosaccharide preparation comprises pentose subunits, hexose subunits, or any combination thereof, wherein each of the said pentose or hexose subunit is independently and optionally functionalized and / or replaced with one of its corresponding anhydro-subunits. In some embodiments, the oligosaccharide preparation comprises hexose subunits, wherein each of the said hexose subunits is independently and optionally replaced with one of its corresponding anhydro-subunits.

[00139] As used herein, a tetrose refers to a monosaccharide with four carbon atoms, such as erythrose, threose, and erythrulose. As used herein, a pentose refers to a monosaccharide with five carbon atoms, such as arabinose, lyxose, ribose, and xylose. As used herein, a hexose refers to a monosaccharide with six carbon atoms, such as allose, altrose, glucose, mannose, gulose, idose, galactose, talose, psicose, fructose, sorbose, and tagatose. As used herein, a heptose refers to a monosaccharide with seven carbon atoms, such as sedoheptulose and mannoheptulose.

[00140] In some embodiments, a herein described oligosaccharide preparation comprises glucose subunit, wherein at least one glucose subunit is optionally replaced with an anhydro-glucose subunit. In some embodiments, a herein described oligosaccharide preparation comprises galactose subunit, wherein at least one galactose subunit is optionally replaced with anhydro- galactose subunit. In some embodiments, a herein described oligosaccharide preparation comprises galactose and glucose subunits, wherein at least one galactose subunit or at least one glucose subunit is optionally replaced with one of its corresponding anhydro-subunits. In some embodiments, a herein described oligosaccharide preparation comprises fructose and glucose subunits, wherein at least one fructose subunit or at least one glucose subunit is optionally replaced with one of its corresponding anhydro-subunits. In some embodiments, a herein described oligosaccharide preparation comprises mannose and glucose subunit, wherein at least one mannose subunit or at least one glucose subunit is optionally replaced with one of its corresponding anhydro-subunits.

[00141] In some embodiments, a herein described oligosaccharide preparation comprises a gluco- galactose-oligosaccharide preparation, a gluco-oligosaccharide preparation, a galacto- oligosaccharide preparation, a fructo-oligosaccharide preparation, a manno-oligosaccharide preparation, an arabino-oligosaccharide preparation, a xylo-oligosaccharide preparation, a gluco- fructo-oligosaccharide preparation, a gluco-manno-oligosaccharide preparation, a gluco-arabino- oligosaccharide preparation, a gluco-xylo-oligosaccharide preparation, a galacto-fructo- oligosaccharide preparation, a galacto-manno-oligosaccharide preparation, a galacto-arabino- oligosaccharide preparation, a galacto-xylo-oligosaccharide preparation, a fructo-manno- oligosaccharide preparation, a fructo-arabino-oligosaccharide preparation, a fructo-xylo- oligosaccharide preparation, a manno-arabino-oligosaccharide preparation, a manno-xylo- oligosaccharide preparation, an arabino-xylo-oligosaccharide preparation, a galacto-arabino-xylo- oligosaccharide preparation, a fructo-galacto-xylo-oligosaccharide preparation, an arabino-fructo- manno-xylo-oligosaccharide preparation, a gluco-fructo-galacto-arabino-oligosaccharide preparation, a fructo-gluco-arabino-manno-xylo oligosaccharide preparation, a gluco-galacto- fructo-manno-arabinoxylo-oligosaccharide preparation, or any combinations thereof; wherein each of the monosaccharide subunit within the preparation is independently and optionally functionalized and / or replaced with one of its corresponding anhydro-subunits.

[00142] In certain embodiments, a herein described oligosaccharide preparation comprises more than 99% of glucose subunits by weight. In some embodiments, the oligosaccharide preparation comprises only glucose subunits.

[00143] In some embodiments, a herein described oligosaccharide preparation comprises about 45% to 55% of glucose subunits and about 55% to 45% of galactose subunits by weight. In some embodiments, the oligosaccharide preparation comprises about 50% glucose and 50% galactose subunits by weight.

[00144] In some embodiments, a herein described oligosaccharide preparation comprises about 80% to 95% of glucose subunits and about 20% to 5% of mannose subunits by weight. In some embodiments, the oligosaccharide preparation comprises about 85 % to 90% of glucose subunits and about 15% to 10% of mannose subunits by weight.

[00145] In some embodiments, a herein described oligosaccharide preparation comprises about 80% to 95% of glucose subunits and about 20% to 5% of galactose subunits by weight. In some embodiments, the oligosaccharide preparation comprises about 85 % to 90% of glucose subunits and about 15% to 10% of galactose subunits by weight.

[00146] In some embodiments, a herein described oligosaccharide preparation comprises about 80% to 95% of glucose subunits, 0% to 8% of galactose subunits, and 5% to 20% of mannose subunits by weight. In some embodiments, the oligosaccharide preparation comprises about 80 % to 90% of glucose subunits, 1% to 5% of galactose subunits, and 10% to 15% of mannose subunits by weight.

[00147] In some embodiments, an oligosaccharide preparation described herein comprises from about 1 wt% to about 100 wt%, from about 50 wt% to about 100 wt%, from about 80 wt% to about 98 wt%, or from about 85 wt% to about 95 wt% of glucose subunits, or any ranges therebetween. In some embodiments, galactose subunits are present in an oligosaccharide preparation described herein at an amount of from about 0 wt% to about 90 wt%, from about 1 wt% to about 50 wt%, from about 2 wt% to about 20wt%, or from about 5 wt% to about 15 wt%, or any ranges therebetween. In some embodiments, mannose subunits are present in an oligosaccharide preparation described herein at an amount of from about 0 wt% to about 90 wt%, from about 1 wt% to about 50 wt%, from about 2 wt% to about 20wt%, or from about 5 wt% to about 15 wt%, or any ranges therebetween. D- vs. L- Form

[00148] In some embodiments, at least one monosaccharide subunit in an oligosaccharide is in L- form. In some embodiments, at least one monosaccharides subunit in an oligosaccharide is in D- form. In some embodiments, the monosaccharide subunits in a herein described oligosaccharide preparation are in their naturally-abundant form, for example, D-glucose, D-xylose, and L- arabinose.

[00149] In some embodiments, a herein described oligosaccharide preparation comprises a mixture of L- and D-forms of monosaccharide subunits. In some embodiments, the ratio of monosaccharide subunits in L- to D- or in D- to L- form is about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:12, about 1:14, about 1:16, about 1:18, about 1:20, about 1:25, about 1:30, about 1:35, about 1:40, about 1:45, about 1:50, about 1:55, about 1:60, about 1:65, about 1:70, about 1:75, about 1:80, about 1:85, about 1:90, about 1:100 or about 1:150. Functionalized Oligosaccharides

[00150] In some embodiments, one or more oligosaccharides in a herein described oligosaccharide preparation are independently functionalized. Functionalized oligosaccharides can be produced by, for example, combining one or more sugars with one or more functionalizing compounds in the presence of a catalyst. Methods of producing functionalized oligosaccharides are described in WO 2012 / 118767, WO 2014 / 031956, and WO / 2016 / 122887.

[00151] In some embodiments, the functionalizing compound comprises one or more acid groups (e.g., -COOH), hydroxyl groups, or N-containing groups (e.g., -CN, -NO2, and -N(Ra)2, wherein Ra is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, aryl, heterocycloalkyl, or heteroaryl groups), S-containing groups (e.g., thiol and sulfates), halides (e.g., -Cl), P- containing groups (e.g., phosphate), or any combination thereof. In some embodiments, the functionalizing compound is linked to at least one monosaccharide subunit via an ether, ester, oxygen-sulfur, amine, or oxygen-phosphorous bond. In some embodiments, one or more functionalizing compounds are linked to a monosaccharide subunit via a single linkage. In some embodiments, at least one functionalizing compound is linked to one or two oligosaccharides via two or more linkages.

[00152] It is to be understood that for each oligosaccharide in the oligosaccharide preparation, each of the described embodiments is independent and can be combined as if each and every combination were listed separately; thus, any combination of the embodiments, are encompassed by the present disclosure. For instance, the various embodiments can be grouped into several categories that include but are not limited to (i) the presence or absence of anhydro-subunit; (ii) the number and level of anhydro-subunit, (iii) the type of species of anhydro-subunit, (iv) the location of anhydro-subunit, (v) the degree of polymerization, (vi) the molecular weight, (vii) the presence or absence of any functional groups, (viii) the type of the oligosaccharide, (ix) the type of glycosidic linkage, and (x) the L- versus D-form. Accordingly, the described oligosaccharide preparation comprises a plurality of oligosaccharides of different species. In some embodiments, In some embodiments, a herein described oligosaccharide preparation comprises at least 10, 102, 103, 104, 105, 106, 107, 108, 109, or 1010 different oligosaccharide species. In some embodiments, the preparation comprises at least 103, 104, 105, 106, or 109 different oligosaccharide species. In some embodiments, the preparation comprises at least 103 different oligosaccharide species. IV. Methods of Manufacturing Oligosaccharide Preparations

[00153] In one aspect, provided herein are methods of manufacturing oligosaccharide preparations. In some embodiments, provided herein are methods of manufacturing oligosaccharide preparations suitable for use in a nutritional composition, such as an animal feed composition, or being fed directly to an animal. In one aspect, provided herein are methods of manufacturing an oligosaccharide preparation, the method comprising heating an aqueous composition comprising one or more feed sugars and a catalyst to a temperature and for a time sufficient to induce polymerization, wherein the catalyst is selected from the group consisting of: (+)-camphor-10-sulfonic acid; 2-pyridinesulfonic acid; 3-pyridinesulfonic acid; 8-hydroxy-5- quinolinesulfonic acid hydrate; <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-hydroxy-2-pyridinemethanesulfonic acid; (<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>)-camphor-10- sulfonic acid; butylphosphonic acid; diphenylphosphinic acid; hexylphosphonic acid; methylphosphonic acid; phenylphosphinic acid; phenylphosphonic acid; tert-butylphosphonic acid; SS)-VAPOL hydrogenphosphate; 6-quinolinesulfonic acid, 3-(1-pyridinio)-1- propanesulfonate; 2-(2-pyridinyl)ethanesulfonic acid; 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine- p,p'-disulfonic acid monosodium salt hydrate; 1,1'-binaphthyl-2,2'-diyl-hydrogenphosphate; bis(4-methoxyphenyl)phosphinic acid; phenyl(3,5-xylyl)phosphinic acid; L-cysteic acid monohydrate; poly(styrene sulfonic acid -co- divinylbenzene); lysine; Ethanedisulfonic acid; Ethanesulfonic acid; Isethionic acid; Homocysteic acid; HEPBS (N-(2-Hydroxyethyl)piperazine- N'-(4-butanesulfonic acid)); HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid); 2- Hydroxy-3-morpholinopropanesulfonic acid; 2-(N-morpholino)ethanesulfonic acid; Methanesulfonic acid; Methaniazide; Naphthalene-1-sulfonic acid; Naphthalene-2-sulfonic acid; Perfluorobutanesulfonic acid; 6-sulfoquinovose; Triflic acid; 2-aminoethanesulfonic acid; Benzoic acid; Chloroacetic acid; Trifluoroacetic acid; Caproic acid; Enanthic acid; Caprylic acid; Pelargonic acid; Lauric acid; Pamitic acid; Stearic acid; Arachidic acid; Aspartic acid; Glutamic acid; Serine; Threonine; Glutamine; Cysteine; Glycine; Proline; Alanine; Valine; Isoleucine; Leucine; Methionine; Phenylalanine; Tyrosine; Tryptophan,, and wherein the oligosaccharide preparation comprises at least n fractions of oligosaccharides each having a distinct degree of polymerization selected from 1 (DP1 fraction) to n (DPn fraction), wherein n is an integer greater than or equal to 2. In some embodiments, n is an integer greater than or equal to 3. In some embodiments, n is an integer within a range of 1 to 100, such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, or 50. In some embodiments, the polymerization of the feed sugars is achieved by a step-growth polymerization. In some embodiments, the polymerization of the feed sugars is achieved by polycondensation. Feed Sugar

[00154] In some embodiments, a method of manufacturing oligosaccharide preparations described herein comprises heating one or more types of feed sugars. In some embodiments, the one or more feed sugars comprise monosaccharides, disaccharides, trisaccharides, tetrasaccharides, or any mixtures thereof.

[00155] In some embodiments, the one or more feed sugars comprise glucose. In some embodiments, the one or more feed sugars comprise glucose and galactose. In some embodiments, the one or more feed sugars comprise glucose, xylose, and galactose. In some embodiments, the one or more feed sugars comprise glucose and mannose. In some embodiments, the one or more feed sugars comprise glucose and fructose. In some embodiments, the one or more feed sugars comprise glucose, fructose, and galactose. In some embodiments, the one or more feed sugars comprise glucose, galactose, and mannose.

[00156] In some embodiments, the one or more feed sugars comprise disaccharides such as lactose, sucrose and cellobiose. In some embodiments, the one or more feed sugars comprise trisaccharides, such as maltotriose or raffinose. In certain embodiments, the one or more feed sugar comprise glucose, mannose, galactose, xylose, malto-dextrin, arabinose, or galactose, or any combinations thereof. In certain embodiments, the one or more feed sugars comprise sugar syrup such as corn syrup. In some embodiments, the one or more feed sugars comprise glucose and lactose. In some embodiments, the one or more feed sugars comprise glucose and sucrose.

[00157] In some embodiments, the type of feed sugars can impact the resulting manufactured oligosaccharide preparations. For example, in some variations where the one or more feed sugars are all glucose, the resulting oligosaccharide preparations comprise gluco-oligosaccharides preparations. In other embodiments, where the one or more feed sugars are all mannose, the resulting oligosaccharide preparations comprise manno-oligosaccharide preparations. In some embodiments, wherein the one or more feed sugars comprise glucose and galactose, the resulting oligosaccharide preparations comprise gluco-galacto-oligosaccharide preparations. In yet other embodiments, where the one or more feed sugars comprise xylose, glucose and galactose, the resulting oligosaccharide preparations comprise gluco-galacto-xylo-oligosaccharide preparations.

[00158] In some embodiments, each of the one or more feed sugars can be independently in its de-hydrate or hydrate form. In some embodiments, the one or more feed sugars comprise glucose, galactose, fructose, mannose, or any combination thereof, and wherein each of the glucose, galactose, fructose, or mannose is independently in its mono-hydrate or de-hydrate form. In some embodiments, the one or more feed sugars comprise a monosaccharide mono-hydrate such as glucose monohydrate. In some embodiments, the one or more feed sugars comprise a saccharide di-hydrate such as trehalose di-hydrate. In some embodiments, the one or more feed sugars comprise at least one sugar in its de-hydrate form and at least one sugar in its hydrate form.

[00159] In some embodiments, the one or more feed sugars can be provided as a sugar solution, in which the sugars are combined with water and fed into the reactor. In some embodiments, the sugars can be fed into the reactor in a solid form and combined with water in the reactor. In some embodiments, the one or more feed sugars are combined and mixed before the addition of water. In some embodiments, the one or more feed sugars are combined into water and mixed thereafter.

[00160] In some embodiments, the method comprises combining two or more feed sugars with the catalyst to produce an oligosaccharide preparation. In some embodiments, the two or more feed sugars comprise from glucose, galactose, fructose, mannose, lactose, or any combination thereof. In some embodiments, the method comprises combining a mixture of sugars (e.g., monosaccharides, disaccharides, and / or trisaccharides) with the catalyst to produce an oligosaccharide preparation. In other embodiments, the method comprises combining a mixture of sugars and sugar alcohols with the catalyst to produce an oligosaccharide preparation.

[00161] In some embodiments, the one or more feed sugars comprise functionalized or modified sugars. Functionalized or modified sugars can comprise amino sugars, sugar acids, sugar alcohols, sugar amides, sugar ethers, or any combination thereof. In some embodiments, amino sugars refer to sugar molecules in which a hydroxyl group is replaced with an amine group. Exemplary amino sugars include, but are not limited to, N-Acetyl-d-glucosamine, mannosamine, neuraminic acid, muramic acid, N-acetyl-neuramin, N-acetyl-muramic, N-acetyl-galactosamine, N-acetyl- mannosa, N-glycolylneuram, acarviosin, D-glucosamine, and D-galactosamine.

[00162] In some embodiments, sugar acids refer to sugars with a carboxyl group. Exemplary sugar acids include, but are not limited to, aldonic acids (such as glyceric acid, xylonic acid, gluconic acid, and ascorbic acid), ulosonic acids (such as neuraminic acid and ketodeoxyoctulosonic acid), uronic acids (such as glucuronic acid, galacturonic acid, and iduronic acid), and aldaric acids (such as tartaric acid, mucic acid, and saccharic acid).

[00163] In some embodiments, sugar alcohols refer to sugar-derived polyols. Exemplary sugar alcohols include, but are not limited to, ethylene glycol, arabitol, glycerol, erythritol, threitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fucitol, iditol, inositol, and volemitol.

[00164] In embodiments, sugar amides refer to sugar molecules that contain a -C(=O)-N- group. In embodiments, sugar ethers refer to sugar molecules that contain an ether bond, such as glucosides.

[00165] In some embodiments, the functionalized or modified sugar acids comprise glucosamine, N-acetylglucosamine, glucuronic acid, galacturonic acid, glucitol, xylitol, mannitol, sorbitol. In some embodiments, the one of more feed sugars comprise deoxysugars, such as fucose, rhamnose, deoxyribose, or fuculose.

[00166] In some embodiments, a herein described method of manufacturing oligosaccharide preparation is performed at gram scale. In some embodiments, a herein described method of manufacturing oligosaccharide preparation is performed at kilogram or higher scale. Accordingly, in some embodiments, the method comprises heating an aqueous composition comprising one or more feed sugars at a quantity of more than 0.5, more than 1, more than 2, more than 3, more than 4, more than 5, more than 6, more than 7, more than 9, more than 10, more than 100, or more than 1000 kg. In some embodiments, the method comprises heating an aqueous composition comprising one or more feed sugars at a quantity of no more than 0.5, 1, 2, 3, 4, 5, 6, 7, 9, 10, 100, 1000, or 1500 kg. In some embodiments, the method comprises heating an aqueous composition comprising one or more feed sugars at a quantity of more than 1 kg. Catalyst

[00167] In some embodiments, a herein described method of manufacturing oligosaccharide preparation comprises the addition of one or more catalysts. In some embodiments, the catalyst provided herein comprises one or more acids. In some embodiments, the catalyst provided herein comprises mineral acid, carboxylic acid; amino acid; sulfonic acid; boronic acid; phosphonic acid; phosphinic acid; sulfuric acid; phosphoric acid; poly(styrene sulfonic acid-co-vinylbenzyl- imidazolium sulfate-co-divinylbenzene); poly(styrene sulfonic acid-co-divinylbenzene); (+)- camphor-10-sulfonic acid; 2-pyridinesulfonic acid; 3-pyridinesulfonic acid; 8-hydroxy-5- quinolinesulfonic acid hydrate; <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-hydroxy-2-pyridinemethanesulfonic acid; (<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>)-camphor-10- sulfonic acid; butylphosphonic acid; diphenylphosphinic acid; hexylphosphonic acid; methylphosphonic acid; phenylphosphinic acid; phenylphosphonic acid; tert-butylphosphonic acid; SS)-VAPOL hydrogenphosphate; 6-quinolinesulfonic acid; 3-(1-pyridinio)-1- propanesulfonate; 2-(2-pyridinyl)ethanesulfonic acid; 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine- p,p'-disulfonic acid monosodium salt hydrate; 1,1'-binaphthyl-2,2'-diyl-hydrogenphosphate; bis(4-methoxyphenyl)phosphinic acid; phenyl(3,5-xylyl)phosphinic acid; L-cysteic acid monohydrate; acetic acid; propionic acid; butanoic acid; glutamic acid; lysine; Ethanedisulfonic acid; Ethanesulfonic acid; Isethionic acid; Homocysteic acid; HEPBS <semantics>(N−(2−<annotation encoding="application / x-tex">(N-(2-< / annotation>< / semantics> Hydroxyethyl)piperazine-N'-(4-butanesulfonic acid)); HEPES (4-(2-hydroxyethyl)-1- piperazineethanesulfonic acid); 2-Hydroxy-3-morpholinopropanesulfonic acid; 2-(N- morpholino)ethanesulfonic acid; Methanesulfonic acid; Methaniazide; Naphthalene-1-sulfonic acid; Naphthalene-2-sulfonic acid; Perfluorobutanesulfonic acid; 6-sulfoquinovose; Triflic acid; 2-aminoethanesulfonic acid; Benzoic acid; Chloroacetic acid; Trifluoroacetic acid; Caproic acid; Enanthic acid; Caprylic acid; Pelargonic acid; Lauric acid; Pamitic acid; Stearic acid; Arachidic acid; Aspartic acid; Glutamic acid; Serine; Threonine; Glutamine; Cysteine; Glycine; Proline; Alanine; Valine; Isoleucine; Leucine; Methionine; Phenylalanine; Tyrosine; Tryptophan; polymeric acid; carbon-supported acid; or any combination thereof.

[00168] In some embodiments, the catalyst provided herein comprises: (+)-camphor-10-sulfonic acid; 2-pyridinesulfonic acid; 3-pyridinesulfonic acid; 8-hydroxy-5-quinolinesulfonic acid hydrate; <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-hydroxy-2-pyridinemethanesulfonic acid; (<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>)-camphor-10-sulfonic acid; butylphosphonic acid; diphenylphosphinic acid; hexylphosphonic acid; methylphosphonic acid; phenylphosphinic acid; phenylphosphonic acid; tert-butylphosphonic acid; SS)-VAPOL hydrogenphosphate; 6-quinolinesulfonic acid, 3-(1-pyridinio)-1-propanesulfonate; 2-(2- pyridinyl)ethanesulfonic acid; 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine-p,p'-disulfonic acid monosodium salt hydrate; 1,1'-binaphthyl-2,2'-diyl-hydrogenphosphate; bis(4- methoxyphenyl)phosphinic acid; phenyl(3,5-xylyl)phosphinic acid; L-cysteic acid monohydrate; poly(styrene sulfonic acid -co- divinylbenzene); lysine; Ethanedisulfonic acid; Ethanesulfonic acid; Isethionic acid; Homocysteic acid; HEPBS (N-(2-Hydroxyethyl)piperazine-N'-(4- butanesulfonic acid)); HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid); 2-Hydroxy- 3-morpholinopropanesulfonic acid; 2-(N-morpholino)ethanesulfonic acid; Methanesulfonic acid; Methaniazide; Naphthalene-1-sulfonic acid; Naphthalene-2-sulfonic acid; Perfluorobutanesulfonic acid; 6-sulfoquinovose; Triflic acid; 2-aminoethanesulfonic acid; Benzoic acid; Chloroacetic acid; Trifluoroacetic acid; Caproic acid; Enanthic acid; Caprylic acid; Pelargonic acid; Lauric acid; Pamitic acid; Stearic acid; Arachidic acid; Aspartic acid; Glutamic acid; Serine; Threonine; Glutamine; Cysteine; Glycine; Proline; Alanine; Valine; Isoleucine; Leucine; Methionine; Phenylalanine; Tyrosine; Tryptophan; or any combination thereof.

[00169] In some embodiments, the catalyst provided herein is (+)-camphor-10-sulfonic acid. In some embodiments, the catalyst provided herein is 2-pyridinesulfonic acid. In some embodiments, the catalyst provided herein is 3-pyridinesulfonic acid. In some embodiments, the catalyst provided herein is 8-hydroxy-5-quinolinesulfonic acid hydrate. In some embodiments, the catalyst provided herein is <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-hydroxy-2-pyridinemethanesulfonic acid. In some embodiments, the catalyst provided herein is <semantics>(β)<annotation encoding="application / x-tex">(\beta)< / annotation>< / semantics>-camphor-10-sulfonic acid. In some embodiments, the catalyst provided herein is butylphosphonic acid. In some embodiments, the catalyst provided herein is diphenylphosphinic acid. In some embodiments, the catalyst provided herein is hexylphosphonic acid. In some embodiments, the catalyst provided herein is methylphosphonic acid. In some embodiments, the catalyst provided herein is phenylphosphinic acid. In some embodiments, the catalyst provided herein is phenylphosphonic acid. In some embodiments, the catalyst provided herein is tert-butylphosphonic acid. In some embodiments, the catalyst provided herein is SS)- VAPOL hydrogenphosphate. In some embodiments, the catalyst provided herein is 6- quinolinesulfonic acid. In some embodiments, the catalyst provided herein is 3-(1-pyridinio)-1- propanesulfonate. In some embodiments, the catalyst provided herein is 2-(2- pyridinyl)ethanesulfonic acid. In some embodiments, the catalyst provided herein is 3-(2-pyridyl)- 5,6-diphenyl-1,2,4-triazine-p,p'-disulfonic acid monosodium salt hydrate. In some embodiments, the catalyst provided herein is 1,1'-binaphthyl-2,2'-diyl-hydrogenphosphate. In some embodiments, the catalyst provided herein is bis(4-methoxyphenyl)phosphinic acid. In some embodiments, the catalyst provided herein is phenyl(3,5-xylyl)phosphinic acid. In some embodiments, the catalyst provided herein is L-cysteic acid monohydrate. In some embodiments, the catalyst provided herein is poly(styrene sulfonic acid -co- divinylbenzene). In some embodiments, the catalyst provided herein is lysine.

[00170] In some embodiments, the catalyst is Ethanedisulfonic acid. In some embodiments, the catalyst is Ethanesulfonic acid. In some embodiments, the catalyst is Isethionic acid. In some embodiments, the catalyst is Homocysteic acid. In some embodiments, the catalyst is HEPBS (N- (2-Hydroxyethyl)piperazine-N'-(4-butanesulfonic acid)). In some embodiments, the catalyst is HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid). In some embodiments, the catalyst is 2-Hydroxy-3-morpholinopropanesulfonic acid. In some embodiments, the catalyst is 2-(N- morpholino) ethanesulfonic acid. In some embodiments, the catalyst is Methanesulfonic acid. In embodiments, the catalyst is Naphthalene-1-sulfonic acid. In some embodiments, the catalyst is some embodiments, the catalyst is Methaniazide. In some Naphthalene-2-sulfonic acid. In some embodiments, the catalyst is Perfluorobutanesulfonic acid. In some embodiments, the catalyst is 6-sulfoquinovose. In some embodiments, the catalyst is Triflic acid. In some embodiments, the catalyst is 2-aminoethanesulfonic acid. In some embodiments, the catalyst is Benzoic acid. In some embodiments, the catalyst is Chloroacetic acid. In some embodiments, the catalyst is Trifluoroacetic acid. In some embodiments, the catalyst is Caproic acid. In some embodiments, the catalyst is Enanthic acid. In some embodiments, the catalyst is Caprylic acid. In some embodiments, the catalyst is Pelargonic acid. In some embodiments, the catalyst is Lauric acid. In some embodiments, the catalyst is Pamitic acid. In some embodiments, the catalyst is Stearic acid. In some embodiments, the catalyst is Arachidic acid. In some embodiments, the catalyst is Aspartic acid. In some embodiments, the catalyst is Glutamic acid. In some embodiments, the catalyst is Serine. In some embodiments, the catalyst is Threonine. In some embodiments, the catalyst is Glutamine. In some embodiments, the catalyst is Cysteine. In some embodiments, the catalyst is Glycine. In some embodiments, the catalyst is Proline. In some embodiments, the catalyst is Alanine. In some embodiments, the catalyst is Valine. In some embodiments, the catalyst is Isoleucine. In some embodiments, the catalyst is Leucine. In some embodiments, the catalyst is Methionine. In some embodiments, the catalyst is Phenylalanine. In some embodiments, the catalyst is Tyrosine. In some embodiments, the catalyst is Tryptophan. In some embodiments, the catalyst provided herein is a polymeric catalyst or a carbon-supported catalyst disclosed in WO 2016122887.

[00171] In some embodiments, the catalyst provided herein is present in an amount of from about 0.01% to about 5%, from about 0.02% to about 4%, from about 0.03% to about 3%, or from about 0.05% to about 2% of the one or more feed sugars by dry weight. In some embodiments, the catalyst provided herein is present in an amount of from about 1% to 2% of the one or more feed sugars by dry weight. In some embodiments, the catalyst provided herein is present in an amount of about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 1.1%, about 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1.6%, about 1.7%, about 1.8%, about 1.9%, about 2.0%, about 2.1%, about 2.2%, about 2.3%, about 2.4%, about 2.5%, about 2.6%, about 2.7%, about 2.8%, about 2.9%, or about 3.0% of the one or more feed sugars by dry weight.

[00172] In some embodiments, the catalyst provided herein is present in an amount of from about 0.01% to about 5%, from about 0.02% to about 4%, from about 0.03% to about 3%, or from about 0.05% to about 2% of the aqueous composition by dry weight. In some embodiments, the catalyst provided herein is present in an amount of from about 1% to 2% of the aqueous composition by dry weight. In some embodiments, the catalyst provided herein is present in an amount of about 0.8%, about 0.9%, about 1.0%, about 1.1%, about 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1.6%, about 1.7%, about 1.8%, about 1.9%, about 2.0%, about 2.1%, about 2.2%, about 2.3%, about 2.4%, about 2.5%, about 2.6%, about 2.7%, about 2.8%, about 2.9%, or about 3.0% of the aqueous composition by dry weight.

[00173] In some embodiments, the catalyst provided herein is a combination of two or more different catalysts. In some embodiments, the catalyst comprises a recyclable catalyst such as resins and polymeric catalysts and a non-recyclable catalyst. In some embodiments, where the catalyst comprises at least two different catalysts, each of the catalyst is present in an amount provided herein. In other embodiments, where the catalyst comprises at least two different catalysts, the at least two different catalysts are present in aggregate in an amount provided herein.

[00174] In some embodiments, the catalyst is added into the aqueous composition in a dry form. In other embodiments, the catalyst is added into the aqueous composition in a wet form such as in an aqueous solution. In some embodiment, the catalyst is combined with the one or more feed sugars before the addition of water. In other embodiments, the catalyst is dissolved into water before its combining with the one or more feed sugars. In some embodiments, the method provided herein comprises producing an aqueous composition by combining the one or more feed sugars in the de-hydrate form and the catalyst in a wet form (e.g., as an aqueous solution). Addition of Water

[00175] In some embodiments, a herein described method of manufacturing oligosaccharide preparations comprises adding water to form an aqueous composition. In some embodiments, all or part of the water in the aqueous composition is added as free water. In other embodiments, all of the water in the aqueous composition is added as bonded water, for example, in saccharide mono- or di-hydrate. In some embodiments, all of the water in the aqueous composition is added as bonded water in monosaccharide mono-hydrate, such as glucose mono-hydrate. In certain embodiments, all or part of the water in the aqueous composition is added with the catalyst, i.e., via a catalyst solution. Water Content

[00176] As the methods of manufacturing the oligosaccharide preparations proceed, water can be produced through reaction. For example, in some embodiments, water is produced (i) with the formation of a glycosidic bond, (ii) with the formation of an anhydro-subunit, or (iii) through other mechanisms or sources. As the sugar condensation and dehydration reactions both involve water, in some embodiments, the water content influences the composition of the oligosaccharide preparation.

[00177] Further, in some embodiments, water content influences the viscosity of the aqueous composition, which in turn can affect the effectiveness of mixing of the aqueous composition. For example, in some embodiments, an overly viscous aqueous composition can lead to an undesirable heterogeneous catalyst distribution in the aqueous composition. Moreover, in some embodiments, very low water content can lead to the solidification of the aqueous composition, which prevents effective mixing. On the other hand, in some other embodiments, exceedingly high water content can impede sugar condensation reaction and lower the level of the anhydro-subunits. Accordingly, the present disclosure describes suitable water content for the manufacturing of oligosaccharide preparations.

[00178] In some embodiments, a herein described method of manufacturing oligosaccharide preparation comprises forming and / or heating an aqueous composition. In some embodiments, the aqueous composition comprises from about 0% to about 80%, from about 0% to about 70%, from about 0% to about 60%, from about 0% to about 50%, from about 0% to about 40%, from about 0% to about 35%, from about 0% to about 30%, from about 0% to about 25%, from about 0% to about 20%, from about 0% to about 19%, from about 0% to about 18%, from about 0% to about 17%, from about 0% to about 16%, from about 0% to about 15%, from about 0% to about 14%, from about 0% to about 13%, from about 0% to about 12%, from about 0% to about 11%, from about 0% to about 10%, from about 0% to about 9%, from about 0% to about 8%, from about 0% to about 7%, from about 0% to about 6%, from about 0% to about 5%, from about 0% to about 4%, from about 0% to about 3%, from about 0% to about 2%, or from about 0% to about 1% of water by total weight. In some embodiments, the aqueous composition comprises from about 1% to about 20%, from about 1% to about 18%, from about 1% to about 16%, from about 1% to about 14%, from about 1% to about 12%, from about 1% to about 10%, from about 1% to about 8%, from about 1% to about 6%, or from about 1% to about 4% of water by total weight. In some embodiments, the aqueous composition comprises from about 3% to about 16%, from about 3% to about 14%, from about 3% to about 12%, from about 3% to about 10%, from about 3% to about 8%, from about 3% to about 6%, from about 5% to about 16%, from about 5% to about 14%, from about 5% to about 12%, from about 5% to about 10%, from about 7% to about 16%, from about 7% to about 14%, from about 7% to about 12%, from about 7% to about 10%, or from about 8% to about 10% of water by total weight. In some embodiments, the aqueous composition comprises about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15% of water by total weight. In some embodiments, the aqueous composition comprises about 9% water by total weight. It should be understood, however, that the amount of water in the aqueous composition can be adjusted based on the reaction conditions and specific catalyst used. In some embodiments, the water content in the aqueous composition as disclosed above is measured at the beginning of the reaction, for example, before heating the feed sugars. In some embodiments, the water content in the aqueous composition as disclosed above is measured at the end of the polymerization or condensation reaction. In some embodiments, the water content in the aqueous composition as disclosed above is measured as an average water content of the beginning of the reaction and at the end of the reaction.

[00179] In certain embodiments, a method described herein can further comprise monitoring the content of water present in the aqueous composition and / or the ratio of water to sugars or catalyst over a period of time. In some embodiments, the method further comprises removing at least a portion of water in the aqueous composition, for example, by distillation. Any method known in the art can be used to remove water from the aqueous composition, including, for example, by vacuum filtration, vacuum distillation, heating, steam, hot air, and / or evaporation.

[00180] In some embodiments, herein described oligosaccharide preparations are hygroscopic. Thus, in some embodiments, the hygroscopicity of the feed sugars and the oligosaccharides formed in the polymerization can affect the rate by which the water can be removed from the aqueous composition.

[00181] In some embodiments, a herein described method comprises removing at least a portion of water in the aqueous composition such that the water content in the aqueous composition is from about 1% to about 20%, from about 1% to about 18%, from about 1% to about 16%, from about 1% to about 14%, from about 1% to about 12%, from about 1% to about 10%, from about 1% to about 8%, from about 2% to about 16%, from about 2% to about 14%, from about 2% to about 12%, from about 2% to about 10%, from about 2% to about 8%, from about 2% to about 6%, from about 4% to about 16%, from about 4% to about 14%, from about 4% to about 12%, from about 4% to about 10%, from about 4% to about 8%, from about 6% to about 16%, from about 6% to about 12%, from about 6% to about 10%, or from about 6% to about 8% by total weight. In some embodiments, the method comprises removing at least a portion of water in the aqueous composition such that the water content in the aqueous composition is from about 2% to about 10%, from about 2% to about 8%, or from about 4% to about 8% by total weight. In some embodiments, the method comprises removing at least a portion of water in the aqueous composition such that the water content in the aqueous composition is about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% by total weight. In some embodiments, the method comprises removing at least a portion of water in the aqueous composition such that the water content in the aqueous composition is from about 4% to about 8 % by total weight. In some embodiments, the method comprises removing at least a portion of water in the aqueous composition such that, at the end of the polymerization and / or condensation reaction, the water content in the aqueous composition is a water content as disclosed above. In some embodiments, the method comprises removing at least a portion of water in the aqueous composition such that, at the beginning of the polymerization and / or condensation reaction, the water content in the aqueous composition is a water content as disclosed above. In some embodiments, the method comprises removing at least a portion of water in the aqueous composition such that, the average water content in the aqueous composition at the beginning and the end of the polymerization and / or condensation reaction is within a range as disclosed above. In some embodiments, the method comprises removing at least a portion of water in the aqueous composition such that, throughout the polymerization and / or condensation reaction, the water content in the aqueous composition remains within a range as disclosed above.

[00182] In some embodiments, a herein described method comprises adding at least a portion of water in the aqueous composition such that the water content in the aqueous composition is from about 1% to about 20%, from about 1% to about 18%, from about 1% to about 16%, from about 1% to about 14%, from about 1% to about 12%, from about 1% to about 10%, from about 1% to about 8%, from about 2% to about 16%, from about 2% to about 14%, from about 2% to about 12%, from about 2% to about 10%, from about 2% to about 8%, from about 2% to about 6%, from about 4% to about 16%, from about 4% to about 14%, from about 4% to about 12%, from about 4% to about 10%, from about 4% to about 8%, from about 6% to about 16%, from about 6% to about 12%, from about 6% to about 10%, or from about 6% to about 8% by total weight. In some embodiments, the method comprises adding at least a portion of water in the aqueous composition such that the water content in the aqueous composition is from about 2% to about 10%, from about 2% to about 8%, or from about 4% to about 8% by total weight. In some embodiments, the method comprises adding at least a portion of water in the aqueous composition such that the water content in the aqueous is about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% by total weight. In some embodiments, the method comprises adding at least a portion of water in the aqueous composition such that the water content in the aqueous composition is from about 4% to about 8% by total weight. In some embodiments, the method comprises adding at least a portion of water in the aqueous composition such that, at the end of the polymerization and / or condensation reaction, the water content in the aqueous composition is a water content as disclosed above. In some embodiments, the method comprises adding at least a portion of water in the aqueous composition such that, at the beginning of the polymerization and / or condensation reaction, the water content in the aqueous composition is a water content as disclosed above. In some embodiments, the method comprises adding at least a portion of water in the aqueous composition such that, the average water content in the aqueous composition at the beginning and the end of the polymerization and / or condensation reaction is within a range as disclosed above. In some embodiments, the method comprises adding at least a portion of water in the aqueous composition such that, throughout the polymerization and / or condensation reaction, the water content in the aqueous composition remains within a range as disclosed above.

[00183] In some embodiments, the degrees of polymerization of the oligosaccharides and / or the amount and type of the anhydro-subunits within the oligosaccharide preparation can be regulated by adjusting or controlling the content of water present in the aqueous composition throughout the manufacturing process. For example, in some embodiments, the degrees of polymerization of the oligosaccharides and the amount of the anhydro-subunits are increased by decreasing the water content.

[00184] Accordingly, in some embodiments, a herein described method comprises in-process control (IPC) of the water content, which can comprise monitoring water content, maintaining water content, increasing water content, decreasing water content, or any combination thereof. In some embodiments, an IPC process comprises maintaining the water content while the aqueous composition is heated to a temperature described herein. In some embodiments, the method comprises maintaining the water content for the time sufficient to induce polymerization. In some embodiments, the method comprises maintaining the water content within a disclosed range by either adding water or removing water from the aqueous composition, or both. In some embodiments, the method comprises maintaining the water content within a disclosed range by distillation. In some embodiments, the method comprises maintaining the water content within a disclosed range by vacuum distillation. In some embodiments, the method comprises maintaining the water content within a disclosed range by distillation under atmosphere pressure.

[00185] In some embodiments, the water content of the aqueous composition is maintained within a range of from about 1% to about 20%, from about 1% to about 18%, from about 1% to about 16%, from about 1% to about 14%, from about 1% to about 12%, from about 1% to about 10%, from about 1% to about 8%, from about 2% to about 16%, from about 2% to about 14%, from about 2% to about 12%, from about 2% to about 10%, from about 2% to about 8%, from about 2% to about 6%, from about 4% to about 16%, from about 4% to about 14%, from about 4% to about 12%, from about 4% to about 10%, from about 4% to about 8%, from about 6% to about 16%, from about 6% to about 12%, from about 6% to about 10%, or from about 6% to about 8% by total weight. In some embodiments, the water content of the aqueous composition is maintained within a range of from about 2% to about 10%, from about 2% to about 8%, or from about 4% to about 8% by total weight. In some embodiments, the water content of the aqueous composition is maintained within a range of from about 2% to about 8% by total weight.

[00186] The water content of the aqueous composition can be determined by a variety of analytical methods and instruments. In some embodiments, the water content is determined by an evaporation method (e.g., loss on drying technique), a distillation method, or a chemical reaction method (e.g., Karl Fischer titration). In some embodiments, the water content is determined by an analytical instrument such as a moisture analyzer. In some embodiments, the water content is determined by Karl Fischer titration.

[00187] In some embodiments, the water content of the aqueous composition is measured during the reaction and used to implement in-process control (IPC) of the water content. In certain embodiments, the water content of the reaction is measured by Karl-Fisher titration, IR spectroscopy, NIR spectroscopy, conductivity, viscosity, density, mixing torque, or mixing energy. In some embodiments, the measurement of the water content of the reaction is used to control an apparatus that actively adjusts the water content of the reaction, such as a water addition pump or flow valve.

[00188] Without being bound by theory, it is believed that water content during the sugar polymerization and / or condensation reaction can affect the level of the anhydro-subunits in a herein described oligosaccharide preparation. For example, as illustrated in FIG. 29, in some embodiments, a higher water content correlates with a lower level of anhydro-subunits. In some embodiments, a lower reaction temperature can correlate with a lower level of anhydro-subunits content. Temperature

[00189] In some embodiments, the degrees of polymerization of the oligosaccharides and / or the amount and type of the anhydro-subunits within the oligosaccharide preparation can be regulated by adjusting the temperature, to which the aqueous composition is heated. In some embodiments, a herein described method of manufacturing an oligosaccharide preparation comprises heating the aqueous composition to a temperature of from about 80 °C to about 250 °C, from about 90°C to about 200 °C, from about 100 °C to about 200 °C, from about 100 °C to about 180 °C, from about 110 °C to about 170 °C, from about 120 °C to about 160 °C, from about 130 °C to about 150 °C, or from about 135 °C to about 145 °C. In some embodiments, the method of manufacturing an oligosaccharide preparation comprises heating the aqueous composition to a temperature of from about 100 °C to about 200 °C, from about 100 °C to about 180 °C, from about 110 °C to about 170 °C, from about 120 °C to about 160 °C, from about 130 °C to about 150 °C, or from about 135 °C to about 145 °C. In some embodiments, the method of manufacturing an oligosaccharide preparation comprises heating the aqueous composition to a temperature of from about 135 °C to about 145 °C. In other embodiments, the method of manufacturing an oligosaccharide preparation comprises heating the aqueous composition to a temperature of from about 125 °C to about 135 °C. Reaction Time

[00190] In some embodiments, a herein described method of manufacturing an oligosaccharide preparation comprises heating the aqueous composition for a sufficient time. In some embodiments, the degrees of polymerization of the oligosaccharides manufactured according to the methods described herein can be regulated by the reaction time.

[00191] In some embodiments, the sufficient time is prescribed by a number of hours. For example, in some embodiments, the sufficient time is at least 30 minutes, at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hour, at least 8 hours, at least 9 hours, or at least 10 hours. In some embodiments, the sufficient time is from about 1 to about 24 hours, from about 1 to about 16 hours, from about 1 to about 8 hours, from about 1 to about 4 hours, from about 1 to about 3 hours, from about 1 to about 2 hours, from about 2 to about 12 hours, from about 2 to about 10 hours, from about 2 to about 8 hours, from about 2 to about 6 hours, from about 2 to about 4 hours, from about 3 to about 8 hours, from about 3 to about 6 hours, from about 3 to about 5 hours, or from about 3 to about 4 hours.

[00192] In some embodiments, the sufficient time is determined by measuring one or more chemical or physical properties of the oligosaccharide preparation, for example, water content, viscosity, molecular weight, anhydro-subunit content, and / or the distribution of degree of polymerization.

[00193] In some embodiments, the molecular weight of the oligosaccharide preparation is monitored during polymerization. In some embodiments, the method comprises heating the aqueous composition for a time sufficient for the aqueous composition to reach a number average molecular weight or weight average molecular weight as described herein. In certain embodiments, the method comprises heating the aqueous composition for a time sufficient for the aqueous composition to reach a number average molecular weight within a range of from about 300 to about 5000 g / mol, from about 500 to about 5000 g / mol, from about 700 to about 5000 g / mol, from about 500 to about 2000 g / mol, from about 700 to about 2000 g / mol, from about 700 to about 1500 g / mol, from about 300 to about 1500 g / mol, from about 300 to about 2000 g / mol, from about 400 to about 1000 g / mol, from about 400 to about 900 g / mol, from about 400 to about 800 g / mol, from about 500 to about 900 g / mol, or from about 500 to about 800 g / mol. In certain embodiments, the method comprises heating the aqueous composition for a time sufficient for the aqueous composition to reach a number average molecular weight of from about 500 to about 2000 g / mol. In certain embodiments, the method comprises heating the aqueous composition for a time sufficient for the aqueous composition to reach a weight average molecular weight within a range of from about 300 to about 5000 g / mol, from about 500 to about 5000 g / mol, from about 700 to about 5000 g / mol, from about 500 to about 2000 g / mol, from about 700 to about 2000 g / mol, from about 700 to about 1500 g / mol, from about 300 to about 1500 g / mol, from about 300 to about 2000 g / mol, from about 400 to about 1300 g / mol, from about 400 to about 1200 g / mol, from about 400 to about 1100 g / mol, from about 500 to about 1300 g / mol, from about 500 to about 1200 g / mol, from about 500 to about 1100 g / mol, from about 600 to about 1300 g / mol, from about 600 to about 1200 g / mol, or from about 600 to about 1100 g / mol. In certain embodiments, the method comprises heating the aqueous composition for a time sufficient for the aqueous composition to reach a weight average molecular weight of from about 700 to about 3000 g / mol.

[00194] In some embodiments, the sufficient time is the time required for the aqueous composition to reach reaction equilibrium at the respective reaction temperature. Accordingly, in some embodiments, the method comprises heating the aqueous composition for a time sufficient for the aqueous composition to reach equilibrium. For example, in some embodiments, the equilibrium is determined by measuring the molecular weight, viscosity, or DP distribution of the aqueous composition.

[00195] In certain embodiments, the equilibrium is determined by measuring the number average or weight average molecular weight of the aqueous composition. In some embodiments, the equilibrium is determined by the number or weight average molecular weight of the aqueous composition that remains essentially unchanged over time. In some embodiments, the equilibrium is determined by a change of the number or weight average molecular weight of the aqueous composition that is less than certain percentage over a period of time. In some embodiments, the molecular weight of the aqueous composition is measured by HPLC or SEC.

[00196] In some embodiments, the equilibrium is determined by a change of the number or weight average molecular weight of the aqueous composition of less than 25%, less than 20%, less than 15%, less than 10%, or less than 5% over a period of time. In some embodiments, the equilibrium is determined by a change of the number or weight average molecular weight of the aqueous composition over a period of 3 hours, 2 hours, 1 hour, 30 minutes, 20 minutes, or 10 minutes. In some embodiments, the equilibrium is determined by a change of the weight average molecular weight of the aqueous composition of less than 15% over the period of 1 hour.

[00197] In certain embodiments, the equilibrium is determined by measuring the viscosity of the aqueous composition. In some embodiments, the equilibrium is determined by the viscosity of the aqueous composition that remains essentially unchanged over time. In some embodiments, the equilibrium is determined by a change of the viscosity of the aqueous composition that is less than certain percentage over a period of time. In some embodiments, the viscosity of the aqueous composition is measured by a viscometer or rheometer.

[00198] In some embodiments, the equilibrium is determined by a change of the viscosity of the aqueous composition of less than 25%, less than 20%, less than 15%, less than 10%, or less than 5% over a period of time. In some embodiments, the equilibrium is determined by a change of the viscosity of the aqueous composition over a period of 3 hours, 2 hours, 1 hour, 30 minutes, 20 minutes, or 10 minutes. In some embodiments, the equilibrium is determined by a change of the viscosity of the aqueous composition of less than 15% over the period of 1 hour.

[00199] In certain embodiments, the equilibrium is determined by measuring the DP distribution of the aqueous composition. In some embodiments, the equilibrium is determined by the DP distribution of the aqueous composition that remains essentially unchanged over time. In some embodiments, a change of the DP distribution of the aqueous composition is determined by calculating a series of Km, wherein <semantics>Km=[DPm][H2O][DPm−1][DP1]<annotation encoding="application / x-tex">Km = \frac{[DP_m][H_2O]}{[DP_{m-1}][DP_1]}< / annotation>< / semantics>, wherein [H2O] represents the molar water concentration (mol / L), and [DP1], [DPm-1], and [DPm] represent the molar concentrations of oligosaccharides (mol / L) in the DP1, DPm-1, and DPm fractions respectively. For example, K2 equals [DP2][H2O] / [DP1][DP1] according to the above formula. In some embodiments, m is an integer larger than 1 and smaller than n. In other embodiments, m equals n. In some embodiment, m is an integer larger than 1 and less than or equal to n. In some embodiments, m is 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[00200] In some embodiments, the concentration of the oligosaccharides in the DP1, DPm-1, and DPm fractions are determined by SEC, HPLC, FFF, A4F, mass spectrometry, or any other suitable method. In some embodiments, the concentration of the oligosaccharides in the DP1, DPm-1, and DPm fractions are determined by SEC such as GPC. In some embodiments, the concentration of the oligosaccharides in the DP1, DPm-1, and DPm fractions are determined by mass spectrometry such as GC-MS, LC-MS / MS, and MALDI-MS. In some embodiments, the concentration of the oligosaccharides in the DP1, DPm-1, and DPm fractions are determined by HPLC. In some embodiments, the water concentration is determined by an evaporation method (e.g., loss on drying technique), a distillation method, or by a chemical reaction method (e.g., Karl Fischer titration). In some embodiments, the water concentration is determined by any suitable analytical instrument such as a moisture analyzer.

[00201] In some embodiments, the method comprises calculating a series of at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 30, at least 40, or at least 50 Km numbers. In some embodiments, the method comprises calculating a series of at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, or at least 15 Km numbers. In some embodiments, the method comprises calculating about 3, 4, 5, 6, 7, 8, 9, 10, or 15 Km numbers. In some embodiments, the method comprises calculating K2 to K4, K2 to K5, K2 to K6, K2 to K7, K2 to K8, K2 to K9, K2 to K10, K2 to K11, K2 to K12, K2 to K13, K2 to K14, K2 to K15, K3 to K5, K3 to K6, K3 to K7, K3 to K8, K3 to K9, K3 to K10, K3 to K11, K3 to K12, K3 to K13, K3 to K14, or K3 to K15. In certain embodiments, the method comprises calculating K2 to K4 or K3 to K5.

[00202] In some embodiments, the value of Km depends on the temperature, water concentration, and / or the amount and type of the feed sugars. In some embodiments, Km is from about 0.1 to about 100, from about 0.1 to about 90, from about 0.1 to about 80, from about 0.1 to about 70, from about 0.1 to about 60, from about 0.1 to about 50, from about 0.1 to about 40, from about 0.1 to about 30, from about 0.1 to about 25, from about 0.1 to about 20, or from about 0.1 to about 15. In some embodiments, Km is from about 1 to about 100, from about 1 to about 90, from about 1 to about 80, from about 1 to about 70, from about 1 to about 60, from about 1 to about 50, from about 1 to about 40, from about 1 to about 30, from about 1 to about 25, from about 1 to about 20, from about 1 to about 15, from about 1 to about 10, from about 5 to about 50, from about 5 to about 40, from about 5 to about 30, from about 5 to about 20, from about 5 to about 15, or from about 5 to about 10. In some embodiments, Km is from about 1 to about 15 or from about 5 to about 15.

[00203] In some embodiments, an average, a standard deviation, and / or a relative standard deviation are determined for the series of Km calculated. As used herein, a relative standard deviation is expressed in percentage, and is obtained by multiplying the standard deviation by 100 and dividing this product by the average.

[00204] In some embodiments, the equilibrium is determined by the relative standard deviation of the series of Km of less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%. In some embodiments, the equilibrium is determined by the relative standard deviation of the series of Km of less than 15%, less than 10%, or less than 5%. Post-Reaction Steps

[00205] In some embodiments, a herein described method of manufacturing oligosaccharide preparations further comprises one or more additional processing steps after heating the aqueous composition at a temperature and for a sufficient time. In some embodiments, the additional processing steps comprise, for example, separation (such as chromatographic separation), dilution, concentration, drying, filtration, demineralization, extraction, decolorization, or any combination thereof. For example, in some embodiments, the method comprises a dilution step and a decolorization step. In some embodiments, the method comprises a filtration step and a drying step.

[00206] In some embodiments, the method comprises a dilution step, where water is added into the oligosaccharide preparation to make a syrup of oligosaccharide preparation. In some embodiments, the concentration of oligosaccharide preparation in the syrup is from about 5% to about 80%, from about 10% to about 70%, from about 10% to about 60%, from about 10% to about 50%, from about 10% to about 40%, from about 10% to about 30%, or from about 15% to about 25%. In other embodiments, the method does not comprise a dilution step, but rather, the oligosaccharide preparation is allowed to solidify. In some embodiments, the method comprises a filtration step. In some embodiments, the method comprises recycling the catalyst by filtration.

[00207] In some embodiments, the method described comprises a decolorization step. In some embodiments, the oligosaccharide preparation can undergo a decolorization step using any method known in the art, including, for example, treatment with an absorbent, activated carbon, chromatography (e.g., using ion exchange resin), hydrogenation, and / or filtration (e.g., microfiltration).

[00208] In some embodiments, the oligosaccharide preparation is contacted with a material to remove salts, minerals, and / or other ionic species. In certain embodiments, the oligosaccharide preparation is flowed through an anionic / cationic exchange column pair. In one embodiment, the anionic exchange column contains a weak base exchange resin in a hydroxide form and the cationic exchange column contains a strong acid exchange resin in a protonated form.

[00209] In some embodiments, the method comprises a concentration step. In some embodiments, the centration step produces an oligosaccharide preparation with increased concentration. For example, in some embodiments, the concentration step comprises evaporation (e.g., vacuum evaporation), drying (e.g., freeze-drying and spray drying) or any combination thereof.

[00210] In some embodiments, the method comprises an isolation step, wherein at least a portion of the oligosaccharide preparation is separated. In some embodiments, the isolation step comprises crystallization, precipitation, filtration (e.g., vacuum filtration), and centrifugation, or any combination thereof.

[00211] In some embodiments, the method comprises a separation step. In some embodiments, the separation step comprises separating at least a portion of the oligosaccharide preparation from at least a portion of the catalyst, from at least a portion of the unreacted feed sugars, or from both. In some embodiments, the separation step comprises filtration, chromatography, differential solubility, precipitation, extraction, or centrifugation. Reactors

[00212] The methods described herein can comprise the use of one or more reactors suitable for sugar condensation, considering the reaction temperature, pH, pressure, and other factors. In some embodiments, the one or more suitable reactors comprise a fed-batch stirred reactor, a batch stirred reactor, a continuous flow stirred reactor, a continuous plug-flow column reactor, an attrition reactor, or a reactor with stirring induced by an electromagnetic field. In some embodiments, the one or more suitable reactors comprise a reactor described in Ryu, S. K., and Lee, J. M., Bioconversion of waste cellulose by using an attrition bioreactor, Biotechnol. Bioeng. 25: 53- 65(1983); Gusakov, A. V., and Sinitsyn, A. P., Kinetics of the enzymatic hydrolysis of cellulose: 1. A mathematical model for a batch reactor process, Enz. Microb. Technol., 7: 346-352 (1985); Gusakov, A. V., Sinitsyn, A. P., Davydkin, I. Y., Davydkin, V. Y., Protas, O. V., Enhancement of enzymatic cellulose hydrolysis using a novel type of bioreactor with intensive stirring induced by electromagnetic field, Appl. Biochem. Biotechnol., 56: 141-153(1996); or Fernanda de Castilhos Corazza, Flavio Faria de Moraes, Gisella Maria Zanin and Ivo Neitzel, Optimal control in fed- batch reactor for the cellobiose hydrolysis, Acta Scientiarum. Technology, 25: 33-38 (2003).

[00213] In some embodiments, the one or more suitable reactors comprise fluidized bed, upflow blanket, immobilized, or extruder type reactors for hydrolysis and / or fermentation. In some embodiments, the one or more suitable reactors comprise an open reactor, a closed reactor, or both. In some embodiments, where the method comprises a continuous process, the one or more suitable reactors can include a continuous mixer such as a screw mixer. Process

[00214] In some embodiments, a herein described method of manufacturing oligosaccharide preparations comprises a batch process, a continuous process, or both. In some embodiments, the method of manufacturing the oligosaccharide preparation comprises a batch process. For example, in some embodiments of the batch process, manufacturing of subsequent batches of the oligosaccharide preparation does not start until the completion of the current batch. In some embodiments, during the batch process, all or a substantial amount of oligosaccharide preparation is removed from the reactor. In some embodiments, during the batch process, all the feed sugars and the catalyst are combined in a reactor before the aqueous composition is heated to the described temperature or before the polymerization is induced. In some embodiments, during the batch process, the feed sugars are added before, after, or simultaneous with the addition of the catalyst.

[00215] In some embodiments, the batch process is a fed-batch process, wherein all the feed sugars are not added into the reactor at the same time. In some embodiments of the fed-batch process, at least a portion of the feed sugars are added into the reactor during polymerization or after the aqueous composition is heated to the described temperature. In some embodiments of the fed-batch process, at least 10%, 20%, 30%, 40%, 50%, or 60% by weight of the feed sugars are added into the reactor during polymerization or after the aqueous composition is heated to the described temperature.

[00216] In some embodiments, the method of manufacturing the oligosaccharide preparation comprises a continuous process. For example, in some embodiments of the continuous process, the contents of the reactor continuously flow through the reactor. In some embodiments, the combination of the feed sugars with the catalyst and the removal of at least a portion of the oligosaccharide preparation are performed concurrently.

[00217] In some embodiments, the method of manufacturing the oligosaccharide preparation comprises a single-pot or multi-pot process. For example, in some embodiments of the single-pot process, the polymerization is performed in a single reactor. For another example, in some embodiments of the multi-pot process, the polymerization is performed in more than one reactor. In some embodiments of the multi-pot process, the method comprises 2, 3, or more reactors. In some embodiments of the multi-pot process, the method comprises a combination step, where the polymerization products from two or more reactors are combined. V. Nutritional Composition Comprising Anhydro-subunit

[00218] Provided herein are nutritional compositions comprising an oligosaccharide preparation. In certain embodiments, provided herein are nutritional compositions comprising a described oligosaccharide preparation, wherein the presence and / or concentration of the oligosaccharide preparation within the nutritional compositions can be selectively determined and / or detected. Oligosaccharide preparations, which exhibit complex functional modulation of a microbial community, can be important components of nutritional compositions. Thus, the presence and / or concentration of an oligosaccharide preparation within nutritional compositions can be one of the factors that need to be measured in the quality control and manufacturing process of the nutritional compositions. Accordingly, the provided nutritional compositions are advantageous in terms of quality control and manufacturing purposes as the presence and / or concentration of the oligosaccharide preparation can be selectively determined and / or detected. For example, in some embodiments, the presence and concentration of the oligosaccharide preparation can be determined and / or detected by measuring a signal associated with the anhydro-subunit containing oligosaccharides.

[00219] In some embodiments, the nutritional composition is an animal feed composition. In some embodiments, the nutritional composition comprises a base nutritional composition. Base Nutritional Compositions

[00220] In some embodiments, a herein described nutritional composition comprises a base nutritional composition and a disclosed oligosaccharide preparation. In some embodiments, the base nutritional composition comprises a carbohydrate source that is different from the oligosaccharide preparation. For example, in some embodiments, the base nutritional composition comprises a naturally occurring carbohydrate source (or naturally occurring oligosaccharide composition) such as starch and plant fibers. In some embodiments, the base nutritional composition comprises starch. In some embodiments, the base nutritional composition comprises plant fibers.

[00221] In some embodiments, the base nutritional composition comprises one or more carbohydrate sources that are derived from: seeds, roots, tubers, corn, tapioca, arrowroot, wheat, rice, potatoes, sweet potato, sago, beans (e.g., favas, lentils, mung beans, peas, and chickpeas.), maize, cassava, or other starchy foods (e.g., acorns, arrowroot, arracacha, bananas, barley, breadfruit, buckwheat, canna, colacasia, katakuri, kudzu, malanga, millet, oats, oca, polynesian arrowroot, sorghum, rye, taro, chestnuts, water chestnuts, and yams).

[00222] In some embodiments, the base nutritional composition comprises one or more carbohydrate sources that are derived from: legumes (e.g., peas, soybeans, lupins, green beans, and other beans), oats, rye, chia, barley, fruits (e.g., figs, avocados, plums, prunes, berries, bananas, apple skin, quinces, and pears), vegetables (e.g., broccoli, carrots, cauliflower, zucchini, celery, nopal, and Jerusalem artichokes), root tubers, root vegetables (e.g., sweet potatoes and onions), psyllium seed husks, seeds (e.g., flax seeds), nuts (e.g., almonds), whole grain foods, wheat, corn bran, lignans, or any combination thereof. In some embodiments, the base nutritional composition comprises one or more plant fibers derived from wheat bran, sugar beet pulp, fuzzy cottonseeds, soy hulls, or any combination thereof.

[00223] In some embodiments, the base nutritional composition comprises less than 500 ppm, less than 400 ppm, less than 300 ppm, less than 200 ppm, less than 100 ppm, less than 50 ppm, less than 10 ppm, less than 5 ppm, or less than 1 ppm anhydro-subunits or anhydro-subunit containing oligosaccharides. In some embodiments, the base nutritional composition comprises less than 50 ppm, less than 10 ppm, less than 5 ppm, or less than 1 ppm anhydro-subunits or anhydro-subunit containing oligosaccharides. In some embodiments, the base nutritional composition is essentially free of anhydro-subunits.

[00224] In some embodiments, the base nutritional composition lacks a detectable level of anhydro-subunits. Depending on the methods of detecting or determination, an anhydro-subunit level below a certain threshold can be undetectable. For example, in some embodiments, a detectable level of anhydro-subunit can refer to at least 1000 ppm, at least 500ppm, at least 400 ppm, at least 300 ppm, at least 200 ppm, at least 100 ppm, at least 50 ppm, at least 10 ppm, at least 5 ppm, or at least 1 ppm of anhydro-subunit or anhydro-subunit containing oligosaccharides in the base nutritional composition.

[00225] In some embodiments, the base nutritional composition comprises a plurality of oligosaccharides. In some embodiments, the base nutritional composition comprises a glycosidic bond type distribution that is different from the oligosaccharide preparation. For example, in some embodiments, the base nutritional composition comprises a higher percentage of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkages than the oligosaccharide preparation. In some embodiments, the glycosidic linkages such as the <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkages in the base nutritional compositions are digestible by one or more enzymes. In some embodiments, the glycosidic linkages in the base nutritional composition are more readily digestible and / or hydrolysable than the glycosidic linkages in the oligosaccharide preparation.

[00226] In some embodiments, the level of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkage, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkage, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkage, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkage in the base nutritional composition is at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, or at least 15% lower than the level of the respective glycosidic linkage in the oligosaccharide preparation. In some embodiments, the level of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkage, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkage, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkage, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkage in the base nutritional composition is at least 10% lower than the level of the respective glycosidic linkage in the oligosaccharide preparation.

[00227] In some embodiments, the level of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkage in the base nutritional composition is at least 50%, at least 40%, at least 35%, at least 30%, at least 25%, at least 20%, at least 15%, at least 10%, at least 5%, or at least 2% higher than the level of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkage in the oligosaccharide preparation. In some embodiments, the level of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkage in the base nutritional composition is at least 10% higher than the level of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkage in the oligosaccharide preparation. Animal Feed Composition

[00228] Depending on the type and age of an animal, a nutritional composition can comprise the oligosaccharide preparation and the base nutritional composition at different ratio. For example, the oligosaccharide preparation can be combined with the base nutritional composition at various ratios suitable for the type and age of an animal. In some embodiments, the oligosaccharide preparation is present in the nutritional composition at a concentration of from about 1 to about 10000 ppm, from about 1 to about 5000 ppm, from about 1 to about 3000 ppm, from about 1 to about 2000 ppm, from about 1 to about 1500 ppm, from about 1 to about 1000 ppm, from about 1 to about 500 ppm, from about 1 to about 250 ppm, from about 1 to about 100 ppm, from about 10 to about 5000 ppm, from about 10 to about 3000 ppm, from about 10 to about 2000 ppm, from about 10 to about 1500 ppm, from about 10 to about 1000 ppm, from about 10 to about 500 ppm, from about 10 to about 250 ppm, from about 10 to about 100 ppm, from about 50 to about 5000 ppm, from about 50 to about 3000 ppm, from about 50 to about 2000 ppm, from about 50 to about 1500 ppm, from about 50 to about 1000 ppm, from about 50 to about 500 ppm, from about 50 to about 250 ppm, from about 50 to about 100 ppm, from about 100 to about 5000 ppm, from about 100 to about 3000 ppm, from about 100 to about 2000 ppm, from about 100 to about 1500 ppm, from about 100 to about 1000 ppm, from about 100 to about 500 ppm, from about 100 to about 400 ppm, from about 100 to about 300 ppm, from about 100 to about 200 ppm, from about 200 to about 5000 ppm, from about 200 to about 3000 ppm, from about 200 to about 2500 ppm, from about 200 to about 2000 ppm, from about 200 to about 1500 ppm, from about 200 to about 1000 ppm, from about 200 to about 500 ppm, from about 500 to about 5000 ppm, from about 500 to about 3000 ppm, from about 500 to about 2500 ppm, from about 500 to about 2000 ppm, from about 500 to about 1500 ppm, or from about 500 to about 1000 ppm. In some embodiments, the oligosaccharide preparation is present in the nutritional composition at a concentration of from about 1 to about 5000 ppm, from about 1 to about 1000 ppm, from about 1 to about 500 ppm, from about 10 to about 5000 ppm, from about 10 to about 2000 ppm, from about 10 to about 1000 ppm, from about 10 to about 500 ppm, from about 10 to about 250 ppm, from about 10 to about 100 ppm, from about 50 to about 5000 ppm, from about 50 to about 2000 ppm, from about 50 to about 1000 ppm, from about 50 to about 500 ppm, from about 50 to about 250 ppm, or from about 50 to about 100 ppm. In some embodiments, the oligosaccharide preparation is present in the nutritional composition at a concentration of from about 1 to about 5000 ppm, from about 10 to about 1000 ppm, from about 10 to about 500 ppm, or from about 50 to about 500 ppm.

[00229] In some embodiments, the oligosaccharide preparation is present in the nutritional composition at a concentration of greater than 10 ppm, greater than 50 ppm, greater than 100 ppm, greater than 200 ppm, greater than 300 ppm, greater than 400 ppm, greater than 500 ppm, greater than 600 ppm, greater than 1000 ppm, or greater than 2000 ppm. In some embodiments, the oligosaccharide preparation is present in the nutritional composition at a concentration of greater than 10 ppm, greater than 50 ppm, greater than 100 ppm, greater than 200 ppm, or greater than 500 ppm.

[00230] In some embodiments, depending on the type and age of an animal, the nutritional composition can further comprise proteins, minerals (such as copper, calcium, and zinc), salts, essential amino acids, vitamins, and / or antibiotics.

[00231] In some embodiments, described herein are methods of manufacturing nutritional compositions for animal feed. In some embodiments, the animal is selected from cattle (e.g., beef cattle and dairy cattle), swine, aquatic animal, poultry, and human. In some embodiments, the animal is swine, such as sows, piglets, and hogs. In other embodiments, the animal is poultry such as chicken, duck, turkey, goose, quail, and hen. In embodiments, the poultry is a broiler, a breeder, or a layer. In some embodiments, the animal is an aquatic animal such salmon, catfish, bass, eel, tilapia, flounder, shrimp, and crab. In some embodiments, the nutritional composition is administered to an animal in a dry form, a liquid form, a paste, or a combination thereof. In some embodiments, the form of administration, the feeding rate, and the feeding schedule can vary depending on the type and age of the animal. Methods of Producing Nutritional Compositions

[00232] Provided herein are methods of manufacturing a nutritional composition comprising: combining an oligosaccharide preparation with a base nutritional composition. In some embodiments, the oligosaccharide preparation comprises anhydro-subunit containing oligosaccharides. In some embodiments, the oligosaccharide preparation comprises a glycosidic bond type distribution that is different from that of the base nutritional composition.

[00233] In some embodiments, the oligosaccharide preparation is a synthetic oligosaccharide preparation. In some embodiments, the synthetic oligosaccharide preparation comprises at least n fractions of oligosaccharides each having a distinct degree of polymerization selected from 1 to n (DP1 to DPn fractions). In some embodiments, n is an integer greater than or equal to 2. In some embodiments, n is an integer greater than 2. In some embodiments, n is an integer greater than or equal to 3. In some embodiments, n is an integer within a range of 1 to 100, such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, or 50. In some embodiments, each of the DP1 to DPn fraction comprises from 0.1% to 90% anhydro-subunit containing oligosaccharides by relative abundance as measured by mass spectrometry. In some embodiments, the DP1 and DP2 fractions of the oligosaccharide preparation each independently comprises from about 0.1% to about 15% or from about 0.5% to about 10% of anhydro-subunit containing oligosaccharides by relative abundance as measured by mass spectrometry. In some embodiments, the DP1 and DP2 fractions of the oligosaccharide preparation each independently comprises anhydro-subunit containing oligosaccharides within a range of from about 0.1%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, or 1.5% to about 8%, 9%, 10%, 11%, 12%, 15% or 20% by relative abundance as measured by mass spectrometry. In some embodiments, the relative abundance of oligosaccharides in each of the n fractions decreases monotonically with its degree of polymerization. In some embodiments, the relative abundance of oligosaccharides in at least 5, 10, 20, or 30 DP fractions decreases monotonically with its degree of polymerization.

[00234] In some embodiments, the method of manufacturing a nutritional composition comprises mixing the oligosaccharide preparation with the base nutritional composition. For example, in some embodiments, the mixing can be performed by an industrial blender and / or mixer such as drum blender, double cone blender, ribbon blender, V blender, shear mixer, and paddle mixer.

[00235] In some embodiments, the method of manufacturing a nutritional composition further comprises a herein described quality control step. In some embodiments, the herein described quality control step comprises determining a level of a signal in a sample of the nutritional composition and calculating a concentration of the oligosaccharide preparation in the nutritional composition based on the level of the signal. In some embodiments, the herein described quality control step comprises detecting a signal in a sample of the nutritional composition through analytical instrumentation, and accepting or rejecting a batch of the nutritional composition based on the presence or absence of the signal. In some embodiments, the herein described quality control step comprises detecting, through analytical instrumentation, the presence or absence of a first signal in a first sample of the nutritional composition, and a second signal in a second sample of the nutritional composition, and comparing the first signal and the second signal. In some embodiments, the signal, the first signal, and / or the second signal is / are (i) indicative of one or more anhydro-subunit containing oligosaccharides, (ii) associated with a degree of polymerization (DP) distribution of oligosaccharides, or (iii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides.

[00236] Additionally, in some embodiments, the method of manufacturing a nutritional composition comprises, after performing the quality control step, further mixing the oligosaccharide preparation with the base nutritional composition, adjusting the level of the oligosaccharide preparation, or a combination thereof. In some embodiments, adjusting the level of the oligosaccharide preparation comprises adding additional oligosaccharide preparation into the nutritional composition or removing a portion of the oligosaccharide preparation from the nutritional composition. In some embodiments, adjusting the level of the oligosaccharide preparation comprises adding additional base nutritional composition into the nutritional composition or removing a portion of the base nutritional composition from the nutritional composition. In some embodiments, adjusting the level of the oligosaccharide preparation comprises adding additional oligosaccharide preparation into the nutritional composition. VI. Method of Correlating Oligosaccharide Preparation

[00237] Provided herein are methods of correlating the presence, absence, and / or concentration of a described oligosaccharide preparation in a nutritional composition. Provided herein are methods of performing a quality control method in the manufacturing process of nutritional compositions comprising an oligosaccharide preparation and a base nutritional composition. Provided herein are methods of determining the quality of nutritional compositions comprising an oligosaccharide preparation and a base nutritional composition. In some embodiments, described herein are methods of correlating a synthetic oligosaccharide preparation in a nutritional composition. In some embodiments, a method of correlating can refer to the establishing, relating, and / or connecting a relationship between two things, and it can also refer to comparing the presence and amount of two things and evaluating a ratio of two things. In some embodiments, described herein are methods of detecting the concentration, the presence, and / or absence of a synthetic oligosaccharide preparation in a nutritional composition. In some embodiments, the nutritional composition comprises a described synthetic oligosaccharide preparation and a naturally occurring oligosaccharide composition (e.g., base nutritional composition).

[00238] In some embodiments, as used herein, the term "quality" can refer to the level of an oligosaccharide preparation (e.g., synthetic oligosaccharide preparation) in the nutritional composition; for example, whether the level is within a specified range such as from 1 to 5000 ppm, 10 to 1000 ppm, 10 to 500 ppm, or 50 to 500 ppm. In some embodiments, the term "quality" can refer to the level of an oligosaccharide preparation in the nutritional composition as evidenced by a signal shown by an analytical instrument; for example, whether a specified peak is present or absent on an NMR, GC-MS, LC-MS / MS, MALDI-MS, or HPLC chromatogram or spectrum and / or a weight determination.

[00239] In some embodiments, the term "quality" can refer to the distribution of an oligosaccharide preparation (e.g., synthetic oligosaccharide preparation) in the nutritional composition, e.g., whether the oligosaccharide preparation is distributed consistently and homogeneously in the nutritional composition. Accordingly, in some embodiments, the quality of a batch of nutritional composition can be determined by comparing the levels and / or the signals of the oligosaccharide preparation in two or more samples of the nutritional composition taken from the same batch. In some embodiments, the quality of a batch of nutritional composition can be determined by comparing the levels and / or the signals of the oligosaccharide preparation in two or more samples of the nutritional composition taken from different batches.

[00240] Thus, provided herein is a method of quantifying an oligosaccharide preparation in a nutritional composition comprising: determining a level of a signal in a sample of the nutritional composition, and calculating a concentration of the oligosaccharide preparation in the nutritional composition based on the level of the signal. Provided herein is a method of performing quality control of a nutritional composition comprising: detecting a signal in a sample of the nutritional composition through analytical instrumentation, and accepting or rejecting a batch of the nutritional composition based on the presence or absence of the signal. Further provided herein is a method of performing quality control of a nutritional composition comprising: detecting, through analytical instrumentation, the presence or absence of a first signal in a first sample of the nutritional composition, and a second signal in a second sample of the nutritional composition, and comparing the first signal and the second signal. In some embodiments, the signal, the first signal, and / or the second signal is / are (i) indicative of one or more anhydro-subunit containing oligosaccharides, (ii) associated with a degree of polymerization (DP) distribution of oligosaccharides, or (iii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides.

[00241] In some embodiments, described herein is a method of quantifying a synthetic oligosaccharide preparation in a nutritional composition, wherein the nutritional composition comprises the synthetic oligosaccharide preparation and a naturally occurring oligosaccharide composition, the method comprising: (a) determining a level of a signal in a sample of the nutritional composition, and (b) correlating a concentration of the oligosaccharide preparation in the nutritional composition based on the level of the signal, wherein the signal is (i) indicative of one or more anhydro-subunit containing oligosaccharides, (ii) associated with a degree of polymerization (DP) distribution of oligosaccharides, or (iii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In some embodiments, described herein is a method of performing quality control of a nutritional composition comprising: (a) providing a batch of a nutritional composition, wherein the nutritional composition comprises a synthetic oligosaccharide preparation and a naturally occurring oligosaccharide composition, (b) obtaining a sample of the nutritional composition from the batch, (c) detecting a signal of at least a portion of oligosaccharides in the sample of the nutritional composition through analytical instrumentation, and (d) accepting or rejecting the batch of the nutritional composition, wherein the signal is (i) indicative of one or more anhydro-subunit containing oligosaccharides, (ii) associated with a degree of polymerization (DP) distribution of oligosaccharides, or (iii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In some embodiments, described herein is a method of performing quality control of a nutritional composition comprising: (a) providing a sample of a nutritional composition, wherein the nutritional composition comprises a naturally occurring oligosaccharide composition, (b) detecting a signal of at least a portion of oligosaccharides in the sample of the nutritional composition through analytical instrumentation, and wherein the signal is indicative of one or more anhydro-subunit containing oligosaccharides, (ii) associated with a degree of polymerization (DP) distribution of oligosaccharides, or (iii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In some embodiments, the signal is indicative of one or more anhydro-subunit containing oligosaccharides. In some embodiments, described herein is a method of performing quality control of a nutritional composition comprising a synthetic oligosaccharide preparation and a naturally occurring oligosaccharide composition, the method comprising: (a) detecting, through analytical instrumentation, the presence or absence of a first signal in a first sample of the nutritional composition, and a second signal in a second sample of the nutritional composition, and (b) comparing the first signal and the second signal, wherein the first signal and the second signal are (i) indicative of one or more anhydro-subunit containing oligosaccharides, (ii) associated with a degree of polymerization (DP) distribution of oligosaccharides, or (iii) associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. In some embodiments, the first signal and / or the second signal are indicative of one or more anhydro-subunit containing oligosaccharides.

[00242] Depending on their types, the signal (e.g., the level of the signal), the first signal, and / or the second signal can be determined or detected by any suitable analytical methods, including without limitation, NMR, HPLC, SEC, FFF, A4F, GC, LC, GC-MS, LC-MS / MS, MALDI-MS, IR, or CD spectroscopy. The level of the signal, the first signal, and / or the second signal can be determined or detected by purifying the sample using LC and weighing the sample. In some embodiments, a signal of a described oligosaccharide preparation is determined or detected by MALDI-MS. In some embodiments, a signal of a described oligosaccharide preparation is determined or detected by LC-MS / MS. In some embodiments, a signal of a described oligosaccharide preparation is determined or detected by GC-MS. In some embodiments, a signal of a described oligosaccharide preparation is determined or detected by NMR such as 2D HSQC NMR. In some embodiments, the relative abundance or the concentration of a described oligosaccharide preparation, or both, are calculated based on the level of a signal of the oligosaccharide preparation, e.g., the relative abundance of one or more peaks on a mass spectrum or the intensity of one or more peaks on an NMR spectrum. Anhydro-subunit Signal

[00243] In some embodiments, a method described herein comprises the detecting of a signal of an oligosaccharide preparation. In some embodiments, a herein described signal of an oligosaccharide preparation is indicative of one or more anhydro-subunit containing oligosaccharides. In some embodiments, a method described herein comprises the detecting of two or more signals, i.e., a first signal, a second signal, etc. In some embodiments, at least one of the first signal and the second signal is indicative of one or more anhydro-subunit containing oligosaccharides. In some embodiments, both the first signal and the second signal are indicative of one or more anhydro-subunit containing oligosaccharides.

[00244] In some embodiments, a signal that is indicative of one or more anhydro-subunit containing oligosaccharides can be determined or detected by any suitable analytical instrumentation, including without limitation, liquid chromatography (such as HPLC), FFF, A4F, NMR, SEC, mass spectrometry such as LC-MS / MS, GC-MS, and MALDI-MS. As used herein, in some embodiments, a signal that is indicative of one or more anhydro-subunit containing oligosaccharides can comprise a signal is attributed to the one or more anhydro-subunit containing oligosaccharides.

[00245] In some embodiments, a signal that is indicative of one or more anhydro-subunit containing oligosaccharides comprises one or more peaks on mass spectrum, NMR, or liquid chromatography (such as HPLC) and / or a weight determination that are attributed to anhydro- subunit containing oligosaccharides. In some embodiments, the one or more peaks on mass spectrum, NMR, or liquid chromatography (such as HPLC) and / or a weight determination are attributed to anhydro-subunit containing oligosaccharides that originate from the oligosaccharide preparation. In some embodiments, the one or more peaks on mass spectrum, NMR, or liquid chromatography (such as HPLC) and / or a weight determination are attributed to anhydro-subunit containing oligosaccharides in any of the DP1 to DPn fraction in the oligosaccharide preparation. In some embodiments, the one or more peaks on mass spectrum, NMR, or liquid chromatography (such as HPLC) and / or a weight determination are attributed to anhydro-subunit containing oligosaccharides in the DP1 fraction. In some embodiments, the one or more peaks on mass spectrum, NMR, or liquid chromatography (such as HPLC) and / or a weight determination are attributed to levoglucosan, 1,6-anhydro-β-D-glucofuranose, or a combination thereof.

[00246] In some embodiments, a signal that is indicative of one or more anhydro-subunit containing oligosaccharides comprises one or more peaks on mass spectrum, NMR, or liquid chromatography (such as HPLC) and / or a weight determination that are attributed to anhydro- subunit containing oligosaccharides in the DP2 fraction. In some embodiments, the one or more peaks on mass spectrum, NMR, or liquid chromatography (such as HPLC) and / or a weight determination are attributed to anhydro-cellobiose. In some embodiments, the one or more peaks on mass spectrum, NMR, or liquid chromatography (such as HPLC) and / or a weight determination are attributed to anhydro-subunit containing oligosaccharides in the DP3, DP4, DP5, or DP6 fraction.

[00247] In some embodiments, the one or more peaks on mass spectrum, NMR, or liquid chromatography (such as HPLC), and / or a weight determination are attributed to anhydro-subunit containing oligosaccharides from more than one fraction. For example, in some embodiments, the signal is attributed to anhydro-subunit containing oligosaccharides from DP1 and DP2 fractions. In some embodiments, the signal is attributed to anhydro-subunit containing oligosaccharides from DP1, DP2, and DP3 fractions.

[00248] For example, in some embodiments, the signal that is indicative of one or more anhydro- subunit containing oligosaccharides comprises one or more peaks on mass spectrum that are attributed to anhydro-subunit containing oligosaccharides in the DP2 fraction. In some embodiments, the signal that is indicative of one or more anhydro-subunit containing oligosaccharides comprises one or more peaks on mass spectrum that are attributed to anhydro- subunit containing oligosaccharides in the DP1 fraction. In some embodiments, the signal that is indicative of one or more anhydro-subunit containing oligosaccharides comprises a weight determination of DP2 anhydro-subunit containing oligosaccharides. In some embodiments, the weight determination is performed for at least a portion of DP2 anhydro-subunit containing oligosaccharides isolated and / or purified by preparative chromatography.

[00249] In some embodiments, a method described herein comprises the detecting of two or more signals, i.e., a first signal, a second signal, a third signal, etc. In some embodiments, the first signal and the second signal are attributed to anhydro-subunit containing oligosaccharides in the same fraction. For example, in some embodiments, the first signal is attributed to levoglucosan and the second signal is attributed to 1,6-anhydro-β-D-glucofuranose. In some embodiments, the first signal and the second signal are attributed to the same species of an anhydro-subunit containing oligosaccharide. For example, in some embodiments, the first signal and the second signal are attributed to 1,6-anhydro-β-D-glucofuranose. In some embodiments, the first signal and the second signal are attributed to anhydro-subunit containing oligosaccharides from different fractions. In some embodiments, the first signal and the second signal are attributed to different species of anhydro-subunit containing oligosaccharides. For example, in some embodiments, the first signal is attributed to levoglucosan and the second signal is attributed to anhydro-cellobiose. In some embodiments, at least one of the first signal and the second signal that are indicative of one or more anhydro-subunit containing oligosaccharides comprises one or more peaks on mass spectrum that are attributed to anhydro-subunit containing oligosaccharides in the DP2 fraction. In some embodiments, at least one of the first signal and the second signal that are indicative of one or more anhydro-subunit containing oligosaccharides comprises one or more peaks on mass spectrum that are attributed to anhydro-subunit containing oligosaccharides in the DP1 fraction. In some embodiments, at least one of the first signal and the second signal that are indicative of one or more anhydro-subunit containing oligosaccharides comprises a weight determination of DP1 and / or DP2 anhydro-subunit containing oligosaccharides. In some embodiments, both the first signal and the second signal that are indicative of one or more anhydro-subunit containing oligosaccharides comprise one or more peaks on mass spectrum that are attributed to anhydro- subunit containing oligosaccharides in the DP2 fraction such as anhydro-cellobiose. In some embodiments, both the first signal and the second signal that are indicative of one or more anhydro-subunit containing oligosaccharides comprise one or more peaks on mass spectrum that are attributed to anhydro-subunit containing oligosaccharides in the DP1 fraction such as levoglucosan, 1,6-anhydro-β-D-glucofuranose, or a combination thereof.

[00250] In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to anhydro-subunit containing oligosaccharides in the DP1 fraction. In some embodiments, among the signal, the first signal, and the second signal, all of them are attributed to anhydro-subunit containing oligosaccharides in the DP1 fraction. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to levoglucosan, 1,6-anhydro-β-D-glucofuranose, or a combination thereof. In some embodiments, among the signal, the first signal, and the second signal, all of them are attributed to levoglucosan, 1,6-anhydro-β-D-glucofuranose, or a combination thereof. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to anhydro-subunit containing oligosaccharides in the DP2 fraction. In some embodiments, among the signal, the first signal, and the second signal, all of them are attributed to anhydro-subunit containing oligosaccharides in the DP2 fraction. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to anhydro- cellobiose. In some embodiments, among the signal, the first signal, and the second signal, all of them are attributed anhydro-cellobiose. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are attributed to anhydro-subunit containing oligosaccharides in the DP3 fraction. In some embodiments, among the signal, the first signal, and the second signal, all of them are attributed to anhydro-subunit containing oligosaccharides in the DP3 fraction.

[00251] In some embodiments, a level of a described signal can be determined by any suitable analytical instrumentation, including without limitation, a balance, liquid chromatography (such as HPLC), NMR, SEC, mass spectrometry such as LC-MS / MS, GC-FID, GC-MS, and MALDI- MS. For example, in some embodiments, the level of a signal is the relative abundance of one or more species of anhydro-subunit containing oligosaccharides represented by the signal. In some embodiments, the level of a signal is the concentration of one or more species of anhydro-subunit containing oligosaccharides represented by the signal. In some embodiments, the level of a signal is the weight of at least a portion of an isolated fraction of the described oligosaccharide preparation (e.g., DP2 anhydro-subunit containing oligosaccharides). Glycosidic Linkages Signal

[00252] In some embodiments, a herein described signal is associated with one or more glycosidic linkages. For example, in some embodiments, the signal is associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>- (1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, or any combination thereof. In some embodiments, the signal is associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages.

[00253] In some embodiments, a method described herein comprises the detecting of two or more signals, i.e., a first signal, a second signal, a third signal, etc. In some embodiments, at least one of the first signal and the second signal is associated with one or more glycosidic linkages. For example, in some embodiments, at least one of the first signal and the second signal is associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, or any combination thereof. In some embodiments, at least one of the first signal and the second signal is associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages.

[00254] In some embodiments, both the first signal and the second signal are associated with one or more glycosidic linkages. For example, in some embodiments, both the first signal and the second signal are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, or any combination thereof. In some embodiments, both the first signal and the second signal are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages.

[00255] In some embodiments, a signal that is associated with one or more glycosidic linkages can be determined or detected by any suitable analytical instrumentation such as NMR, including without limitation, 1D 1H NMR, 1D 13C NMR, 2D NMR such as 2D JRES, HSQC, DOSY, HMBC, COSY, ECOSY, TOCSY, NOESY, or ROESY, or any combination thereof. In some embodiments, a signal that is associated with one or more glycosidic linkages can refer that the signal is attributed to the one or more glycosidic linkages.

[00256] In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, all of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, all of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, all of them are associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, one or more of them are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1, 2) glycosidic linkages. In some embodiments, among the signal, the first signal, and the second signal, all of them are associated with <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1, 2) glycosidic linkages. In some embodiments, among the signal, the first signal,...

Claims

<pat:ClaimStatement>CLAIMS< / pat:ClaimStatement> <pat:Claims com:id="claims"> <pat:Claim com:id="CLM-00001"> <pat:ClaimNumber>1< / pat:ClaimNumber> <pat:ClaimText>1. A method of correlating a synthetic oligosaccharide preparation in a nutritional composition, wherein the nutritional composition comprises the synthetic oligosaccharide preparation and a naturally occurring oligosaccharide composition, the method comprising: a. providing a sample of the nutritional composition, b. detecting a signal of at least a portion of oligosaccharides in the sample of the nutritional composition, and c. correlating a concentration of the synthetic oligosaccharide preparation in the nutritional composition, wherein the signal is indicative of one or more anhydro-subunit containing oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00002"> <pat:ClaimNumber>2< / pat:ClaimNumber> <pat:ClaimText>2. A method of performing quality control of a nutritional composition comprising: a. providing a batch of a nutritional composition, wherein the nutritional composition comprises a synthetic oligosaccharide preparation and a naturally occurring oligosaccharide composition, b. obtaining a sample of the nutritional composition from the batch, c. detecting a signal of at least a portion of oligosaccharides in the sample of the nutritional composition through analytical instrumentation, and d. accepting or rejecting the batch of the nutritional composition, wherein the signal is indicative of one or more anhydro-subunit containing oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00003"> <pat:ClaimNumber>3< / pat:ClaimNumber> <pat:ClaimText>3. The method of claim 1 or 2, wherein the one or more anhydro-subunit containing oligosaccharides have a degree of polymerization of 2 (DP2). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00004"> <pat:ClaimNumber>4< / pat:ClaimNumber> <pat:ClaimText>4. A method of performing quality control of a nutritional composition comprising: a. providing a sample of a nutritional composition, wherein the nutritional composition comprises a naturally occurring oligosaccharide composition, and b. detecting a signal of at least a portion of oligosaccharides in the sample of the nutritional composition through analytical instrumentation, wherein the signal is indicative of one or more anhydro-subunit containing oligosaccharides having a degree of polymerization of 2 (DP2). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00005"> <pat:ClaimNumber>5< / pat:ClaimNumber> <pat:ClaimText>5. The method of claim 4, wherein the nutritional composition comprises a synthetic oligosaccharide preparation. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00006"> <pat:ClaimNumber>6< / pat:ClaimNumber> <pat:ClaimText>6. The method of claim 2 or 5, comprising correlating a concentration of the synthetic oligosaccharide preparation in the nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00007"> <pat:ClaimNumber>7< / pat:ClaimNumber> <pat:ClaimText>7. The method of claim 1 or 2, wherein the signal is indicative of anhydro-subunit containing oligosaccharides having a degree of polymerization of 1 (DP1). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00008"> <pat:ClaimNumber>8< / pat:ClaimNumber> <pat:ClaimText>8. The method of claim 7, wherein the signal is attributed to levoglucosan, 1,6-anhydro-β-D- glucofuranose, or a combination thereof. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00009"> <pat:ClaimNumber>9< / pat:ClaimNumber> <pat:ClaimText>9. The method of claim 1 or 2, wherein the signal is indicative of anhydro-subunit containing oligosaccharides having a degree of polymerization of 3 (DP3). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00010"> <pat:ClaimNumber>10< / pat:ClaimNumber> <pat:ClaimText>10. The method of claim 1 or 2, wherein the signal is indicative of DP1, DP2, or DP3 anhydro- subunit containing oligosaccharides, or a combination thereof. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00011"> <pat:ClaimNumber>11< / pat:ClaimNumber> <pat:ClaimText>11. The method of claim 1 or 2, wherein the signal is attributed to anhydro-cellobiose. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00012"> <pat:ClaimNumber>12< / pat:ClaimNumber> <pat:ClaimText>12. The method of any one of claims 1 to 3 or 5 to 11, wherein the one or more anhydro-subunit containing oligosaccharides originate from the synthetic oligosaccharide preparation. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00013"> <pat:ClaimNumber>13< / pat:ClaimNumber> <pat:ClaimText>13. The method of any one of claims 1 to 12, wherein the signal is detected by high- performance liquid chromatography (HPLC), gas chromatography (GC), mass spectrometry (MS), nuclear magnetic resonance (NMR) spectroscopy, size exclusion chromatography (SEC), field flow fractionation (FFF), asymmetric flow field flow fractionation (A4F), weight determination of fractions by preparative chromatography, or any combination thereof. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00014"> <pat:ClaimNumber>14< / pat:ClaimNumber> <pat:ClaimText>14. The method of any one of claims 1 to 13, wherein the nutritional composition comprises a base nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00015"> <pat:ClaimNumber>15< / pat:ClaimNumber> <pat:ClaimText>15. The method of claim 14, wherein the base nutritional composition comprises the naturally occurring oligosaccharide composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00016"> <pat:ClaimNumber>16< / pat:ClaimNumber> <pat:ClaimText>16. The method of claim 14 or 15, wherein the base nutritional composition lacks a detectable level of anhydro-subunit containing oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00017"> <pat:ClaimNumber>17< / pat:ClaimNumber> <pat:ClaimText>17. The method of any one of claims 14 to 16, wherein the base nutritional composition is essentially free of anhydro-subunit containing oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00018"> <pat:ClaimNumber>18< / pat:ClaimNumber> <pat:ClaimText>18. The method of any one of claims 1 to 17, wherein the detecting comprises a weight determination of one or more degree of polymerization (DP) fractions of oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00019"> <pat:ClaimNumber>19< / pat:ClaimNumber> <pat:ClaimText>19. The method of any one of claims 1 to 18, wherein the detecting comprises a weight determination of at least a portion of anhydro-subunit containing oligosaccharides from the sample. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00020"> <pat:ClaimNumber>20< / pat:ClaimNumber> <pat:ClaimText>20. The method of claim 19, wherein the at least a portion of anhydro-subunit containing oligosaccharides have a degree of polymerization of 1, 2, or 3. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00021"> <pat:ClaimNumber>21< / pat:ClaimNumber> <pat:ClaimText>21. The method of any one of claims 1 to 17, wherein the detecting comprises a weight determination of one or more degree of polymerization (DP) fractions of oligosaccharides and / or a weight determination of at least a portion of anhydro-subunit containing oligosaccharides from the sample, and the one or more DP fractions of oligosaccharides or the at least a portion of anhydro-subunit containing oligosaccharides are isolated by preparative chromatography, and / or wherein the at least a portion of anhydro-subunit containing oligosaccharides have a degree of polymerization of 1, 2, or 3. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00022"> <pat:ClaimNumber>22< / pat:ClaimNumber> <pat:ClaimText>22. The method of any one of claims 1 to 21, wherein the signal is detected, at least in part, by matrix-assisted laser desorption / ionization-mass spectrometry (MALDI-MS). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00023"> <pat:ClaimNumber>23< / pat:ClaimNumber> <pat:ClaimText>23. The method of any one of claims 1 to 21, wherein the signal is detected, at least in part, by liquid chromatography—mass spectrometry (LC-MS) / MS. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00024"> <pat:ClaimNumber>24< / pat:ClaimNumber> <pat:ClaimText>24. The method of any one of claims 1 to 21, wherein the signal is detected, at least in part, by GC-flame ionization detector (GC-FID) or gas chromatography-mass spectrometry (GC- MS). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00025"> <pat:ClaimNumber>25< / pat:ClaimNumber> <pat:ClaimText>25. The method of any one of claims 1 to 21, wherein the signal is detected, at least in part, by NMR spectroscopy. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00026"> <pat:ClaimNumber>26< / pat:ClaimNumber> <pat:ClaimText>26. The method of any one of claims 1 to 25, wherein the detecting comprises determining the presence or absence of the signal. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00027"> <pat:ClaimNumber>27< / pat:ClaimNumber> <pat:ClaimText>27. The method of claim 26, wherein the detecting comprises determining the presence or absence of DP1 or DP2 anhydro-subunit containing oligosaccharides, or both. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00028"> <pat:ClaimNumber>28< / pat:ClaimNumber> <pat:ClaimText>28. The method of any one of claims 1 to 27, wherein the detecting comprises determining or correlating a level of the signal. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00029"> <pat:ClaimNumber>29< / pat:ClaimNumber> <pat:ClaimText>29. The method of claim 28, wherein the detecting comprises correlating a level of DP1 anhydro-subunit containing oligosaccharides in the nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00030"> <pat:ClaimNumber>30< / pat:ClaimNumber> <pat:ClaimText>30. The method of claim 28 or 29, wherein the detecting comprises correlating a level of DP2 anhydro-subunit containing oligosaccharides in the nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00031"> <pat:ClaimNumber>31< / pat:ClaimNumber> <pat:ClaimText>31. A method of performing quality control of a nutritional composition comprising a synthetic oligosaccharide preparation and a naturally occurring oligosaccharide composition, the method comprising: a. providing a first sample of the nutritional composition, b. providing a second sample of the nutritional composition, c. detecting a first signal of at least a portion of oligosaccharides in the first sample, d. detecting a second signal of at least a portion of oligosaccharides in the second sample, and e. comparing the first signal and the second signal, wherein the first signal and the second signal are independently indicative of one or more anhydro-subunit containing oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00032"> <pat:ClaimNumber>32< / pat:ClaimNumber> <pat:ClaimText>32. The method of claim 31, comprising correlating a concentration of the synthetic oligosaccharide preparation in the nutritional composition of the first sample, a concentration of the synthetic oligosaccharide preparation in the nutritional composition of the second sample, or both. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00033"> <pat:ClaimNumber>33< / pat:ClaimNumber> <pat:ClaimText>33. The method of claim 32, wherein the first signal and the second signal are attributed to the same species of an anhydro-subunit containing oligosaccharide. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00034"> <pat:ClaimNumber>34< / pat:ClaimNumber> <pat:ClaimText>34. The method of claim 32, wherein the first signal and the second signal are attributed to different species of anhydro-subunit containing oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00035"> <pat:ClaimNumber>35< / pat:ClaimNumber> <pat:ClaimText>35. The method of any one of claims 31 to 34, wherein the first sample and the second sample are taken from different batches of the nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00036"> <pat:ClaimNumber>36< / pat:ClaimNumber> <pat:ClaimText>36. The method of any one of claims 31 to 35, wherein the first signal and the second signal are each independently detected by high-performance liquid chromatography (HPLC), gas chromatography (GC), mass spectrometry (MS), nuclear magnetic resonance (NMR) spectroscopy, size exclusion chromatography (SEC), field flow fractionation (FFF), asymmetric flow field flow fractionation (A4F), weight determination of fractions by preparative chromatography, or any combination thereof. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00037"> <pat:ClaimNumber>37< / pat:ClaimNumber> <pat:ClaimText>37. The method of any one of claims 31 to 36, wherein the nutritional composition comprises a base nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00038"> <pat:ClaimNumber>38< / pat:ClaimNumber> <pat:ClaimText>38. The method of claim 37, wherein the base nutritional composition comprises the naturally occurring oligosaccharide composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00039"> <pat:ClaimNumber>39< / pat:ClaimNumber> <pat:ClaimText>39. The method of claim 37 or 38, wherein the base nutritional composition lacks a detectable level of anhydro-subunit containing oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00040"> <pat:ClaimNumber>40< / pat:ClaimNumber> <pat:ClaimText>40. The method of any one of claims 37 to 39, wherein the base nutritional composition is essentially free of anhydro-subunit containing oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00041"> <pat:ClaimNumber>41< / pat:ClaimNumber> <pat:ClaimText>41. The method of any one of claims 31 to 40, wherein the one or more anhydro-subunit containing oligosaccharides originate from the synthetic oligosaccharide preparation. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00042"> <pat:ClaimNumber>42< / pat:ClaimNumber> <pat:ClaimText>42. The method of any one of claims 31 to 41, wherein the first signal, the second signal, or both are independently attributed to anhydro-subunit containing oligosaccharides having a degree of polymerization of 1 (DP1). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00043"> <pat:ClaimNumber>43< / pat:ClaimNumber> <pat:ClaimText>43. The method of claim 42, wherein the first signal, the second signal, or both are independently attributed to levoglucosan, 1,6-anhydro-β-D-glucofuranose, or a combination thereof. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00044"> <pat:ClaimNumber>44< / pat:ClaimNumber> <pat:ClaimText>44. The method of any one of claims 31 to 41, wherein the first signal, the second signal, or both are independently attributed to anhydro-subunit containing oligosaccharides having a degree of polymerization of 2 (DP2). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00045"> <pat:ClaimNumber>45< / pat:ClaimNumber> <pat:ClaimText>45. The method of claim 44, wherein the signal is attributed to anhydro-cellobiose. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00046"> <pat:ClaimNumber>46< / pat:ClaimNumber> <pat:ClaimText>46. The method of any one of claims 31 to 41, wherein the first signal, the second signal, or both are independently attributed to anhydro-subunit containing oligosaccharides having a degree of polymerization of 3 (DP3). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00047"> <pat:ClaimNumber>47< / pat:ClaimNumber> <pat:ClaimText>47. The method of any one of claims 31 to 41, wherein the first signal, the second signal, or both are independently attributed to DP1, DP2, or DP3 anhydro-subunit containing oligosaccharides, or a combination thereof. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00048"> <pat:ClaimNumber>48< / pat:ClaimNumber> <pat:ClaimText>48. The method of any one of claims 31 to 47, wherein the detecting comprises a weight determination of one or more degree of polymerization (DP) fractions from the sample or the second sample. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00049"> <pat:ClaimNumber>49< / pat:ClaimNumber> <pat:ClaimText>49. The method of any one of claims 31 to 48, wherein the detecting comprises a weight determination of at least a portion of anhydro-subunit containing oligosaccharides from the first sample or the second sample. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00050"> <pat:ClaimNumber>50< / pat:ClaimNumber> <pat:ClaimText>50. The method of claim 49, wherein the at least a portion of anhydro-subunit containing oligosaccharides have a degree of polymerization of 1, 2, or 3. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00051"> <pat:ClaimNumber>51< / pat:ClaimNumber> <pat:ClaimText>51. The method of any one of claims 48 to 50, wherein the one or more DP fractions of oligosaccharides or the at least a portion of anhydro-subunit containing oligosaccharides are isolated by preparative chromatography. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00052"> <pat:ClaimNumber>52< / pat:ClaimNumber> <pat:ClaimText>52. The method of any one of claims 31 to 51, wherein the first signal, the second signal, or both are independently detected, at least in part, by matrix-assisted laser desorption / ionization-mass spectrometry (MALDI-MS). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00053"> <pat:ClaimNumber>53< / pat:ClaimNumber> <pat:ClaimText>53. The method of any one of claims 31 to 51, wherein the first signal, the second signal, or both are independently detected, at least in part, by liquid chromatography-mass spectrometry (LC-MS) / MS. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00054"> <pat:ClaimNumber>54< / pat:ClaimNumber> <pat:ClaimText>54. The method of any one of claims 31 to 51, wherein the first signal, the second signal, or both are independently detected, at least in part, by GC-flame ionization detector (GC-FID) or gas chromatography-mass spectrometry (GC-MS). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00055"> <pat:ClaimNumber>55< / pat:ClaimNumber> <pat:ClaimText>55. The method of any one of claims 31 to 51, wherein the first signal, the second signal, or both are independently detected, at least in part, by NMR spectroscopy. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00056"> <pat:ClaimNumber>56< / pat:ClaimNumber> <pat:ClaimText>56. The method of any one of claims 31 or 36 to 55, wherein the first signal is indicative of one or more anhydro-subunit containing oligosaccharides and the second signal is associated with <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,6) glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics> glycosidic linkages, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,1)-<semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics> glycosidic linkages of oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00057"> <pat:ClaimNumber>57< / pat:ClaimNumber> <pat:ClaimText>57. The method of any one of claims 31 to 56, wherein the detecting comprises determining the presence or absence of the first signal and the second signal. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00058"> <pat:ClaimNumber>58< / pat:ClaimNumber> <pat:ClaimText>58. The method of claim 57, wherein the detecting comprises determining the presence or absence of DP1 or DP2 anhydro-subunit containing oligosaccharides, or both. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00059"> <pat:ClaimNumber>59< / pat:ClaimNumber> <pat:ClaimText>59. The method of any one of claims 31 to 56, wherein the detecting comprises determining or correlating a level of the first signal and the second signal. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00060"> <pat:ClaimNumber>60< / pat:ClaimNumber> <pat:ClaimText>60. The method of claim 59, wherein the detecting comprises correlating a level of DP1 anhydro-subunit containing oligosaccharides in the nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00061"> <pat:ClaimNumber>61< / pat:ClaimNumber> <pat:ClaimText>61. The method of claim 59 or 60, wherein the detecting comprises correlating a level of DP2 anhydro-subunit containing oligosaccharides in the nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00062"> <pat:ClaimNumber>62< / pat:ClaimNumber> <pat:ClaimText>62. The method of any one of claims 1, 4, 5 or 31 to 61, further comprising accepting or rejecting a batch of the nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00063"> <pat:ClaimNumber>63< / pat:ClaimNumber> <pat:ClaimText>63. The method of any one of claims 1 to 3 or 31 to 61, further comprising adjusting a level of the synthetic oligosaccharide preparation in the nutritional composition after the detecting. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00064"> <pat:ClaimNumber>64< / pat:ClaimNumber> <pat:ClaimText>64. The method of any one of claims 8 to 11 or 37 to 40, wherein the base nutritional composition comprises a plurality of oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00065"> <pat:ClaimNumber>65< / pat:ClaimNumber> <pat:ClaimText>65. The method of claim 64, wherein the base nutritional composition comprises starch or plant fibers. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00066"> <pat:ClaimNumber>66< / pat:ClaimNumber> <pat:ClaimText>66. The method of any one of the claims 8 to 11, 37 to 40 or 64 to 65, wherein a level of <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>- (1,2) glycosidic linkage, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-(1,3) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,2) glycosidic linkage, <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,3) glycosidic linkage, or <semantics>β<annotation encoding="application / x-tex">\beta< / annotation>< / semantics>-(1,4) glycosidic linkage in the base nutritional composition is at least 10% lower than a level of the same glycosidic linkage in the synthetic oligosaccharide preparation. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00067"> <pat:ClaimNumber>67< / pat:ClaimNumber> <pat:ClaimText>67. The method of any one of claims 1 to 66, comprising a derivatization step prior to the detecting. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00068"> <pat:ClaimNumber>68< / pat:ClaimNumber> <pat:ClaimText>68. The method of any one of claims 1 to 67, further comprising extracting oligosaccharides from the sample of the nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00069"> <pat:ClaimNumber>69< / pat:ClaimNumber> <pat:ClaimText>69. The method of claim 68, further comprising filtering or clarifying the extracted oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00070"> <pat:ClaimNumber>70< / pat:ClaimNumber> <pat:ClaimText>70. The method of claim 68 or 69, further comprising concentrating the extracted oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00071"> <pat:ClaimNumber>71< / pat:ClaimNumber> <pat:ClaimText>71. The method of claim 70, wherein the concentrating comprises nano-filtration. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00072"> <pat:ClaimNumber>72< / pat:ClaimNumber> <pat:ClaimText>72. The method of claim 70, wherein the concentrating comprises freeze-drying. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00073"> <pat:ClaimNumber>73< / pat:ClaimNumber> <pat:ClaimText>73. The method of any one of claims 70 to 72, further comprising introducing an internal standard into the extracted or concentrated oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00074"> <pat:ClaimNumber>74< / pat:ClaimNumber> <pat:ClaimText>74. The method of any one of claims 70 to 73, further comprising reducing the extracted or concentrated oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00075"> <pat:ClaimNumber>75< / pat:ClaimNumber> <pat:ClaimText>75. The method of any one of the claims 70 to 73, further comprising digesting the extracted or concentrated oligosaccharides with one or more hydrolytic enzymes. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00076"> <pat:ClaimNumber>76< / pat:ClaimNumber> <pat:ClaimText>76. The method of claim 75, wherein the one or more hydrolytic enzymes comprise carbohydratase, protease, lipase, or any combination thereof. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00077"> <pat:ClaimNumber>77< / pat:ClaimNumber> <pat:ClaimText>77. The method of claim 75 or 76, wherein the one or more hydrolytic enzymes comprise <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>- amylase, amyloglycosidase, invertase, <semantics>α<annotation encoding="application / x-tex">\alpha< / annotation>< / semantics>-galactosidase, or any combination thereof. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00078"> <pat:ClaimNumber>78< / pat:ClaimNumber> <pat:ClaimText>78. The method of any one of claims 75 to 77, wherein the one or more hydrolytic enzymes cleave one or more naturally occurring glycosidic bonds. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00079"> <pat:ClaimNumber>79< / pat:ClaimNumber> <pat:ClaimText>79. The method of any one of claims 75 to 78, further comprising isolating the oligosaccharides that are not digested. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00080"> <pat:ClaimNumber>80< / pat:ClaimNumber> <pat:ClaimText>80. The method of any one of claims 75 to 78, further comprising separating the extracted, concentrated, or digested oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00081"> <pat:ClaimNumber>81< / pat:ClaimNumber> <pat:ClaimText>81. The method of claim 74, further comprising separating the extracted, concentrated, or reduced oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00082"> <pat:ClaimNumber>82< / pat:ClaimNumber> <pat:ClaimText>82. The method of claim 80 or 81, wherein the oligosaccharides are separated chromatographically. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00083"> <pat:ClaimNumber>83< / pat:ClaimNumber> <pat:ClaimText>83. The method of claim 80 or 81, wherein the oligosaccharides are separated by nano- filtration. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00084"> <pat:ClaimNumber>84< / pat:ClaimNumber> <pat:ClaimText>84. The method of claim 82 or 83, further comprising isolating the separated oligosaccharides. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00085"> <pat:ClaimNumber>85< / pat:ClaimNumber> <pat:ClaimText>85. The method of claim 84, wherein the oligosaccharides are separated or isolated by their degrees of polymerization. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00086"> <pat:ClaimNumber>86< / pat:ClaimNumber> <pat:ClaimText>86. The method of claim 85, the method comprises isolating or separating the oligosaccharides with a degree of polymerization of 1, 2, 3, 4, or 5. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00087"> <pat:ClaimNumber>87< / pat:ClaimNumber> <pat:ClaimText>87. The method of claim 86, wherein the DP1 oligosaccharides are isolated or separated. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00088"> <pat:ClaimNumber>88< / pat:ClaimNumber> <pat:ClaimText>88. The method of claim 86, wherein the DP2 oligosaccharides are isolated or separated. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00089"> <pat:ClaimNumber>89< / pat:ClaimNumber> <pat:ClaimText>89. The method of claim 86, wherein the DP3 oligosaccharides are isolated or separated. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00090"> <pat:ClaimNumber>90< / pat:ClaimNumber> <pat:ClaimText>90. The method of any one of claims 1 to 86, comprising isolating at least a portion of the DP1 anhydro-subunit containing oligosaccharides from the sample. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00091"> <pat:ClaimNumber>91< / pat:ClaimNumber> <pat:ClaimText>91. The method of any one of claims 1 to 86, comprising isolating at least a portion of the DP2 anhydro-subunit containing oligosaccharides from the sample. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00092"> <pat:ClaimNumber>92< / pat:ClaimNumber> <pat:ClaimText>92. The method of any one of claims 1 to 86, comprising isolating at least a portion of the DP3 anhydro-subunit containing oligosaccharides from the sample. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00093"> <pat:ClaimNumber>93< / pat:ClaimNumber> <pat:ClaimText>93. The method of any one of claims 84 to 89, wherein the isolated or separated oligosaccharides are quantified. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00094"> <pat:ClaimNumber>94< / pat:ClaimNumber> <pat:ClaimText>94. The method of any one of claims 90 to 92, wherein the isolated oligosaccharides are quantified. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00095"> <pat:ClaimNumber>95< / pat:ClaimNumber> <pat:ClaimText>95. The method of claim 93 or 94, wherein a majority of the quantified oligosaccharides originate from the synthetic oligosaccharide preparation. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00096"> <pat:ClaimNumber>96< / pat:ClaimNumber> <pat:ClaimText>96. The method of any one of claims 93 to 95, wherein more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, or more than 99% by weight of the quantified oligosaccharide originate from the synthetic oligosaccharide preparation. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00097"> <pat:ClaimNumber>97< / pat:ClaimNumber> <pat:ClaimText>97. A method of correlating a synthetic oligosaccharide preparation in a nutritional composition, wherein the nutritional composition comprises (i) a synthetic oligosaccharide preparation that comprises anhydro-subunit containing oligosaccharides and (ii) a naturally occurring oligosaccharide composition, the method comprising: a. providing a sample of the nutritional composition, b. isolating one or more anhydro-subunit containing oligosaccharides from the sample, c. detecting a signal that is indicative of the one or more anhydro-subunit containing oligosaccharides, wherein the detecting comprises (i) a weight determination of at least a portion of anhydro-subunit containing oligosaccharides from the sample or (ii) analyzing at least a portion of anhydro-subunit containing oligosaccharides from the sample by matrix-assisted laser desorption / ionization-mass spectrometry (MALDI-MS), liquid chromatography—mass spectrometry (LC-MS) / MS, or gas chromatography (GC)- MS, and d. correlating a concentration of the synthetic oligosaccharide preparation in the nutritional composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00098"> <pat:ClaimNumber>98< / pat:ClaimNumber> <pat:ClaimText>98. The method of claim 97, wherein the detecting comprises a weight determination of at least a portion of anhydro-subunit containing oligosaccharides having a degree of polymerization of 1 (DP1) from the sample. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00099"> <pat:ClaimNumber>99< / pat:ClaimNumber> <pat:ClaimText>99. The method of claim 97, wherein the detecting comprises a weight determination of at least a portion of anhydro-subunit containing oligosaccharides having a degree of polymerization of 2 (DP2) from the sample. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00100"> <pat:ClaimNumber>100< / pat:ClaimNumber> <pat:ClaimText>100. The method of claim 97, wherein the detecting comprises analyzing at least a portion of DP1 or DP2 anhydro-subunit containing oligosaccharides from the sample by matrix- assisted laser desorption / ionization-mass spectrometry (MALDI-MS). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00101"> <pat:ClaimNumber>101< / pat:ClaimNumber> <pat:ClaimText>101. The method of claim 97, wherein the detecting comprises analyzing at least a portion of DP1 or DP2 anhydro-subunit containing oligosaccharides from the sample by liquid chromatography—mass spectrometry (LC-MS) / MS. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00102"> <pat:ClaimNumber>102< / pat:ClaimNumber> <pat:ClaimText>102. The method of claim 97, wherein the detecting comprises analyzing at least a portion of DP1 or DP2 anhydro-subunit containing oligosaccharides from the sample by gas chromatography / mass spectrometry (GC / MS). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00103"> <pat:ClaimNumber>103< / pat:ClaimNumber> <pat:ClaimText>103. The method of claim 97, wherein the detecting comprises analyzing at least a portion of DP3 anhydro-subunit containing oligosaccharides from the sample by weight determination, matrix-assisted laser desorption / ionization-mass spectrometry (MALDI- MS), liquid chromatography—mass spectrometry (LC-MS) / MS, or gas chromatography- mass spectrometry (GC-MS). < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00104"> <pat:ClaimNumber>104< / pat:ClaimNumber> <pat:ClaimText>104. The method of any one of claims 97 to 103, wherein the isolating comprises separating the one or more anhydro-subunit containing oligosaccharides by preparative chromatography. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00105"> <pat:ClaimNumber>105< / pat:ClaimNumber> <pat:ClaimText>105. The method of any one of claims 1 to 3, 31 to 61, 63 or 97 to 104, wherein the synthetic oligosaccharide preparation is present in the nutritional composition at a concentration of from about 1 to about 5000 ppm, from about 1 to about 1000 ppm, from about 1 to about 500 ppm, from about 10 to about 5000 ppm, from about 10 to about 2000 ppm, from about 10 to about 1000 ppm, from about 10 to about 500 ppm, from about 10 to about 250 ppm, from about 10 to about 100 ppm, from about 50 to about 5000 ppm, from about 50 to about 2000 ppm, from about 50 to about 1000 ppm, from about 50 to about 500 ppm, from about 50 to about 250 ppm, or from about 50 to about 100 ppm. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00106"> <pat:ClaimNumber>106< / pat:ClaimNumber> <pat:ClaimText>106. The method of any one of the claims 1 to 3, 31 to 61, 63 or 97 to 104, wherein the synthetic oligosaccharide preparation is present in the nutritional composition at a concentration of greater than 10 ppm, greater than 50 ppm, greater than 100 ppm, greater than 200 ppm, greater than 300 ppm, greater than 400 ppm, greater than 500 ppm, greater than 600 ppm, greater than 1000 ppm, or greater than 2000 ppm. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00107"> <pat:ClaimNumber>107< / pat:ClaimNumber> <pat:ClaimText>107. The method of any one of the claims 1 to 106, wherein the nutritional composition is an animal feed composition. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00108"> <pat:ClaimNumber>108< / pat:ClaimNumber> <pat:ClaimText>108. A method of manufacturing a nutritional composition comprising: a. combining a base nutritional composition with a synthetic oligosaccharide preparation comprising anhydro-subunit containing oligosaccharides, and b. performing a quality control method as provided in any one of claims 2, 4 to 6 and 31 to 61. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00109"> <pat:ClaimNumber>109< / pat:ClaimNumber> <pat:ClaimText>109. The method of claim 108, wherein the synthetic oligosaccharide preparation comprises at least n fractions of oligosaccharides each having a distinct degree of polymerization selected from 1 to n (DP1 to DPn fractions), wherein n is an integer greater than or equal to 3; and wherein the DP1 and DP2 fractions each independently comprises from about < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00110"> <pat:ClaimNumber>110< / pat:ClaimNumber> <pat:ClaimText>110. The method of claim 109, wherein the relative abundance is determined by liquid chromatography—mass spectrometry (LC-MS) / MS. < / pat:ClaimText> < / pat:Claim> <pat:Claim com:id="CLM-00111"> <pat:ClaimNumber>111< / pat:ClaimNumber> <pat:ClaimText>111. The method of claim 109 or 110, wherein the relative abundance of oligosaccharides in each of the n fractions decreases monotonically with degree of polymerization of the respective fraction. < / pat:ClaimText> < / pat:Claim> < / pat:Claims>