Unsubstituted modified starch and preparation method thereof

Abstract

An unsubstituted crosslinked starch has a hot paste viscosity at pH 3 and 5.5 wt% solids in the range of 100 cP to 1500 cP, a retrogradation enthalpy at 1 week or less and 7 J / g or less at 4 weeks, and a retrogradation end temperature of less than 75 DEG C. The starch may be prepared from waxy cassava, waxy rice, waxy sacchar-2 corn mutant, short-chain waxy potato, waxy wheat, highly phosphorylated waxy starch, or a combination of any two or more thereof. The starch may be cross-linked. The starch may be used in food products, nutritional products, pharmaceutical products, personal care products, paper products, and the like. A method of making starch includes mixing native starch with water to form a slurry; adjusting the pH of the slurry to be alkaline; and mixing a cross-linking agent with the slurry to realize a cross-linking reaction.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 492,269, filed on March 27, 2023, which is incorporated herein by reference in its entirety. Technical Field

[0003] The present disclosure relates to modified starches. Specifically, the present disclosure relates to unsubstituted cross-linked waxy starches and to methods of making such modified starches. Background Art

[0004] Starch typically contains two types of polymers: essentially linear amylose and branched amylopectin. Starches low in amylose are often referred to as "waxy" starch. Exemplary sources of waxy starch include waxy cassava, waxy rice, the waxy sugar-2 corn mutant, short-chain waxy potatoes, waxy wheat, and highly phosphorylated waxy starches. In nature, root and tuber starches typically have higher amounts of phosphate monoesters covalently bound to the starch (ranging from 50 ppm to 980 ppm). Cereal starches such as maize, rice, and wheat, on the other hand, typically have lower amounts of phosphate monoesters (typically ranging from 0 ppm to 80 ppm). Starch phosphorylation, catalyzed by proteins with glucan water dikinase (GWD) or phospho-glucan water dikinase (PWD) activity, creates additional phosphate monoester bonds in the glucan chains. Covalently bound phosphate monoesters can be enhanced through selective breeding programs or genetic engineering.

[0005] Starch can be used to thicken (e.g., increase viscosity) foods and other products, such as cosmetics and pharmaceutical products. Starches from different sources differ in texture, flavor, and viscosity-increasing properties. Native starch can be modified to better control the properties provided by the starch and improve the stability of the starch. Such properties include, for example, viscosity, texture (smoothness, non-cohesiveness), processing stability, cold storage stability, etc. Typically, substitution with various alkyl or hydroxyalkyl groups (e.g., acetyl or hydroxypropyl) has been used to provide cold storage stability and increased viscosity. Cross-linking has been used to control processing stability and maintain viscosity.

[0006] Improvements in modified starches and methods of making them are desirable. Summary of the Invention

[0007] Modified starches, and specifically unsubstituted crosslinked waxy starches, are provided. In one aspect, the technology provides an unsubstituted crosslinked starch having a hot paste viscosity in the range of 100 cP to 1500 cP (measured at pH 3 and 5.5 wt% solids) and a retrogradation enthalpy measured at 4°C for 1 week, and a gelatinization enthalpy, wherein the retrogradation enthalpy is about 25% or less, 20% or less, 15% or less, 10% or less, or 5% or less of the gelatinization enthalpy. The unsubstituted crosslinked starch can have a retrogradation enthalpy of 2.3 J / g or less measured at 4°C for 1 week. The unsubstituted crosslinked starch can have a retrogradation enthalpy of 50% or less of the gelatinization enthalpy measured at 4°C for 4 weeks.

[0008] In one aspect, the technology provides an unsubstituted crosslinked starch that can have a hot paste viscosity in the range of about 100 cP to about 1500 cP measured at pH 3 and 5.5 wt% solids. The unsubstituted crosslinked starch can have a retrogradation enthalpy of 3 J / g or less at 1 week and 7 J / g or less at 4 weeks, and a retrogradation end temperature of less than 75°C. The unsubstituted crosslinked starch can have a hot paste viscosity in the range of about 300 cP to about 1300 cP, about 400 cP to about 1200 cP, about 500 cP to about 1200 cP, about 600 cP to about 1200 cP, about 600 cP to about 1000 cP, about 400 cP to about 900 cP, or about 600 cP to about 900 cP. The unsubstituted crosslinked starch can have a retrogradation enthalpy of about 2.8 J / g or less, about 2.5 J / g or less, about 2.4 J / g or less, about 2.3 J / g or less, about 2.2 J / g or less, about 2.1 J / g or less, or about 2.0 J / g or less measured at 1 week of cold storage. The unsubstituted crosslinked starch can have a retrogradation enthalpy of about 0.7 J / g or more, about 0.8 J / g or more, about 0.9 J / g or more, or about 1.0 J / g or more measured at 1 week of cold storage. At 4 weeks of cold storage, the unsubstituted crosslinked starch can have a retrogradation enthalpy of about 7 J / g or less, about 6.7 J / g or less, about 6.5 J / g or less, about 6.3 J / g or less, about 6.2 J / g or less, 6.1 J / g or less, or about 6.0 J / g or less.

[0009] The unsubstituted cross-linked starch can be prepared from any waxy starch source, including waxy cassava, waxy rice, waxy sugar-2 corn mutant, short chain waxy potato, waxy wheat, highly phosphorylated waxy starch, or a combination of any two or more thereof. The unsubstituted cross-linked starch can be cross-linked. In some aspects, the unsubstituted cross-linked starch can comprise or can have unsubstituted cross-linked waxy cassava starch. The unsubstituted cross-linked starch can be cross-linked using any suitable cross-linking agent, such as sodium trimetaphosphate, phosphorous oxychloride, adipate salt, epichlorohydrin, citric acid, or a combination of any two or more thereof.

[0010] The unsubstituted cross-linked starch can comprise about 5% or less amylose by weight. In some aspects, the unsubstituted cross-linked starch can be free or substantially free of amylose.

[0011] The unsubstituted cross-linked starch can be prepared from a cassava plant, including but not limited to a cassava plant modified using CRISPER / Cas9 technology. Additionally or alternatively, in any aspect herein, the unsubstituted cross-linked starch can be prepared from a naturally occurring waxy cassava plant or a waxy cassava plant developed through non-genetic modification or conventional breeding.

[0012] According to some aspects of the present technology, the unsubstituted cross-linked starch is not hydroxypropylated or acetylated.

[0013] According to some aspects of the present technology, the unsubstituted cross-linked starch can be granular (unpregelatinized).

[0014] The unsubstituted cross-linked starch can be stable in 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 8 or more, or 10 or more freeze-thaw cycles. According to some aspects of the present technology, the unsubstituted cross-linked starch is stable in 4 or more freeze-thaw cycles.

[0015] In another aspect, the present technology provides a pregelatinized non-granular or partially gelatinized or agglomerated starch prepared using the unsubstituted cross-linked starch described herein. In another aspect, the present technology provides a food product comprising the unsubstituted cross-linked starch. The food product can be a dairy product. The dairy product can be or can comprise a yogurt, a sour cream, a fruit topping, a dairy-based dessert, a pudding, or a combination of any two or more thereof. The food product can be a non-dairy yogurt or a non-dairy pudding. The food product can be a sauce, a gravy, a sauce, a dip, a spread, a baked good filling, or a dry mix. The food product can be a pet food, a retort pouch food, or a canned pet food.

[0016] The food product can include unsubstituted cross-linked starch at a rate of about 1 wt% or greater, about 2 wt% or greater, about 5 wt% or greater, about 7.5 wt% or greater, about 10 wt% or greater, about 12.5 wt% or greater, about 15 wt% or greater, or about 20 wt% or greater, based on the total weight of the food product. The unsubstituted cross-linked starch can be included at about 35 wt% or less, about 30 wt% or less, about 25 wt% or less, about 20 wt% or less, about 15 wt% or less, or about 10 wt% or less, based on the total weight of the food product. In some aspects, the unsubstituted cross-linked starch comprises about 1 wt% to about 35 wt%, or about 5 wt% to about 25 wt% of the food product.

[0017] In another aspect, the present technology provides a method of making unsubstituted cross-linked starch as described herein in any aspect. The method can include mixing native starch with water to form a slurry; adjusting the pH of the slurry to an alkaline pH; and mixing a cross-linking agent with the slurry to effect a cross-linking reaction. The resulting unsubstituted cross-linked starch exhibits a hot paste viscosity in the range of about 100 cP to about 1500 cP (measured at pH 3 and 5.5 wt% solids) and a retrogradation enthalpy measured at 4°C for 1 week, and a gelatinization enthalpy, wherein the retrogradation enthalpy is about 25% or less, 20% or less, 15% or less, 10% or less, or 5% or less of the gelatinization enthalpy. The resulting unsubstituted cross-linked starch can exhibit a retrogradation enthalpy of about 2.3 J / g or less at 1 week and about 7 J / g or less at 4 weeks, and a retrogradation end temperature of less than about 75°C.

[0018] The slurry can include about 20 wt% to about 45 wt% native starch dry solids. The pH of the slurry can be adjusted to a range of about 10 to about 13. The cross-linking reaction can be conducted at a reaction temperature of about 24°C to about 45°C and have a duration of 0.5 h to about 10 h.

[0019] The cross-linking agent added to the slurry can include sodium trimetaphosphate, phosphorous oxychloride, adipate, epichlorohydrin, citric acid, or a combination of any two or more thereof. The method can further include mixing a salt into the slurry. The slurry can include 0.5 wt% to 10 wt% of the salt.

[0020] The method can further include adjusting the pH to a range of 5 to 6 after cross-linking.

[0021] The method can further include dewatering and drying the starch.

[0022] Native starches can include waxy tapioca, waxy rice, waxy sugar-2 corn mutant, short chain waxy potato, waxy wheat, highly phosphorylated waxy starch, or combinations of any two or more thereof. Native starches can comprise 5% or less by weight amylopectin. In some aspects, native starches are free or substantially free of amylopectin.

[0023] According to some aspects of the technology, the native starch can comprise waxy tapioca starch, and the unsubstituted crosslinked starch comprises unsubstituted crosslinked waxy tapioca starch.

[0024] The unsubstituted crosslinked starch can exhibit a hot paste viscosity in the range of 400 cP to 1300 cP.

[0025] The method can be free of hydroxypropylation or acetylation. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1A is a graphical representation of the RVA viscosity profile (pH 3 buffer / 5.5% DS) for the POCI3 crosslinked sample of Example 1.

[0027] Figure 1B is a graphical representation of the RVA viscosity profile (pH 6.5 buffer / 5.5% DS) for the POCI3 crosslinked sample of Example 1.

[0028] Figure 1C is a graphical representation of the RVA viscosity profile (pH 3 buffer / 5.5% DS) for the STMP crosslinked sample of Example 1.

[0029] Figure 1D is a graphical representation of the RVA viscosity profile (pH 6.5 buffer / 5.5% DS) for the STMP crosslinked sample of Example 1.

[0030] Figure 1E is a graphical representation of the DSC retrogradation enthalpy results for the samples of Example 1.

[0031] Definitions

[0032] Unless defined otherwise, all scientific and technical terms used herein have the meanings commonly understood by one of ordinary skill in the art. Definitions provided herein are to facilitate understanding of certain terms frequently used herein and are not meant to limit the scope of the disclosure.

[0033] The term "tapioca starch" is used herein to refer to starch obtained from the roots of the cassava plant (Manihot esculenta).

[0034] The term "waxy starch" is used herein to refer to a starch with a low amylose content, such as a starch comprising 5% by weight or less, 3% by weight or less, 2% by weight or less, or no (i.e., free of) amylose.

[0035] The term "gelatinization" is used herein to refer to the phase transition of starch upon heating in excess water, when semi-crystalline starch granules undergo a change of state from an ordered structure to a disordered structure.

[0036] The term "retrogradation" refers to the re-association of disordered glucan chains of gelatinized starch into an ordered structure via hydrogen bonds. Throughout the rearrangement process, the intermolecular spacing between glucan chains decreases resulting in water removal from the gel, a phenomenon known as "syneresis". Starch retrogradation, including the retrogradation enthalpy and the melting transition temperature, can be characterized using differential scanning calorimetry (DSC). The retrogradation enthalpy reflects the melting of crystallites or the unwinding of double helices of amylopectin of retrograded starch. The melting transition temperature is described as the onset temperature (To) and the end temperature (Tc). The melting temperature range reflects the temperature between the onset and end temperature of the melting of crystallites. The end temperature Tc can be referred to as the end of retrogradation temperature or the end of the retrogradation temperature. The measurement of the retrogradation enthalpy To and Tc is also discussed in Vamadevan and Bertoft, Impact of Different Structural Types of Amylopectin on Retrogradation, Food Hydrocolloids 80 (2018) 88.

[0037] The term gelatinization enthalpy is used to refer to the melting of starch crystallites (loss of double helix order or breaking of H-bonds between glucan chains). Differential scanning calorimetry (DSC) is widely used to detect the phase transition. DSC measures the gelatinization transition temperatures (onset [To], peak [Tp] and end [Tc]) and the gelatinization enthalpy (AH). The gelatinization enthalpy (AH) is estimated by integrating the area between the thermogram curve and the baseline under the peak, and expressed as J / g dry starch. The gelatinization enthalpy refers to the amount of energy required to gelatinize starch, i.e., to melt the crystalline structure of starch (order to disorder state). The gelatinization enthalpy and its measurement are also discussed in Vamadevan et al., On the Importance of Organization of Glucan Chains on Thermal Properties of Starch, Carbohydrate Polymers 92 (2013) 1653.

[0038] The terms "stable" and "stability" are generally used to refer to the ability to maintain structure, texture, and / or viscosity. In the context of starch, the terms "stable" and "stability" can be used to refer to various aspects of stability, including processing stability under elevated heat and shear (shear conditions) and cold storage stability. Chemically crosslinked starches provide a desirable smooth texture and have viscosity stability throughout processing operations. Lack of processing stability can be inferred from loss of viscosity and development of poor texture during processing (e.g., under acidic or high shear conditions). Processing stability can include resistance to processing conditions such as heat, acid, shear stress, etc. Cold storage stability can include both cold temperature stability and freeze-thaw stability (stability during freeze-thaw cycles). Cold temperature stability can be determined by measuring the gelatinization enthalpy of the starch after storage at cold temperature (e.g., 1 week or 4 weeks). Lower gelatinization enthalpy indicates improved cold temperature stability. Freeze-thaw stability can be determined by exposing the starch to multiple freeze-thaw cycles and observing changes in the starch (e.g., water separation). Higher number of cycles without changes indicates improved freeze-thaw stability.

[0039] The term "cold temperature" is used herein to refer to the temperature range of about 2 °C to about 6 °C typically used in refrigerators. When used in the context of stability testing, the term "cold temperature" is used to mean about 4 °C.

[0040] The term "thickening starch" is used to refer to a starch that is capable of imparting increased viscosity.

[0041] The term "highly phosphorylated" is used herein to refer to a plant variety that contains at least 10% more, at least 20% more, or at least 30% more phosphomonoesters than a traditional variety. The traditional variety of the plant includes phosphomonoesters that are naturally produced during starch metabolism in the plant without selective breeding or overexpression of GWD through transgenic methods.

[0042] Unless otherwise indicated, the terms "polymer" and "polymeric material" include, but are not limited to, organic homopolymers, copolymers (such as, for example, block, graft, random, and alternating copolymers, terpolymers, and the like), and blends and modifications thereof. Furthermore, unless specifically limited otherwise, the term "polymer" shall include all possible geometric configurations of the material. These configurations include, but are not limited to, isotactic, syndiotactic, and atactic symmetries.

[0043] The term "alkylated" is used in the present disclosure to describe a compound that has reacted to replace a hydrogen atom or a negative charge of the compound with an alkyl group, such that the alkyl group is covalently bonded to the compound.

[0044] The term“alkyl” is used in the present disclosure to describe a monovalent group that is a radical of an alkane and includes straight-chain, branched, cyclic, and bicyclic alkyl groups as well as combinations thereof, including both unsubstituted alkyl groups and substituted alkyl groups. Unless otherwise indicated, alkyl groups typically contain 1 to 30 carbon atoms. In any aspect herein, an alkyl group contains 1 to 20 carbon atoms, 1 to 10 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms. Generally, an alkyl group is attached to the rest of the molecule by a single bond, e.g., an alkyl group can include, but is not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, i-butyl, t-butyl, i-propyl, n-octyl, n-heptyl, ethylhexyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like. In some aspects, an alkyl group is optionally substituted with one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, -OR a , -SR a , -OC(O)-R b , -N(R a )2, -C(O)OR a , -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , N(R a )S(O)2 R b , -S(O)2O R a , and -S(O)-2N(R a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl, and each R b is independently hydrogen, alkyl, fluoroalkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl.

[0045] As used herein, the term“substantially” has the same meaning as“significantly” and can be understood to modify the following term by at least about 95%, at least about 98%, at least about 99%, or at least about 99.5%. The term“substantially free of’ a particular compound means that a composition of the present disclosure contains less than 0.1% of the stated compound.

[0046] As used herein, the term“substantially not” has the same meaning as“not significantly” and can be understood to have the opposite meaning of“substantially,” i.e., to modify the following term by no more than 10%, no more than 5%, or no more than 2%.

[0047] Also, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading the document and is not to be interpreted as limiting; information that is relevant to a section heading can occur within or outside of that particular section. Any publication, patent, or patent document that is cited is incorporated by reference in its entirety to the same extent as if each such document was specifically and individually indicated to be incorporated by reference.

[0048] As used herein, the terms “for example,” “e.g.,” or “such as” are meant to introduce examples that further clarify more general subject matter. Unless otherwise noted, these examples are provided only as an aid to understanding the applications illustrated in the present disclosure, and are not intended to be limiting in any way.

[0049] In the methods described herein, the acts can be carried out in the particular order listed herein. Alternatively, any of the aspects disclosed herein can be carried out in any order without departing from the principles of the present disclosure, unless expressly maintained otherwise by a temporal or operational sequence. Moreover, specific acts can be performed concurrently, unless otherwise indicated by explicit claim language or the inherent meaning of a claim. For example, an act of performing X claimed and an act of performing Y claimed can be performed concurrently in a single operation, and the resulting process would fall within the literal scope of the claimed process.

[0050] The term “about” is used herein in connection with a quantity to include normal fluctuations that would be expected by persons skilled in the art to occur in a measurement, and is understood to have the same meaning as “approximately” and encompasses a typical error range, such as ±10%, ±5%, or ±1% of the stated value or range limit, and includes the exact stated value or range.

[0051] Terms such as “one” and “the” and similar referents are not intended to be singular unless the context clearly indicates otherwise. It is to be understood that the terms “a” or “an” can mean one or more than one, depending upon the context in which it is used. Stated simply, any term that is used in singular form can include its plural counterpart, and vice versa, unless the context clearly indicates otherwise.

[0052] The terms “one” and “the” are used interchangeably with the term “at least one.” The phrases “at least one of’ and “comprises at least one of’ followed by a list of two or more items mean that any of the items can be present, and also that a combination of any two or more of the items can be present.

[0053] As used herein, the term “or” is generally used in its usual sense, including “and / or”, unless the content clearly dictates otherwise. The term “and / or” means one or all of the listed elements or a combination of any two or more of the listed elements.

[0054] A range of values expressed in a terminal manner includes all values included in the range (e.g., 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.80, 4, 5, etc., or 10 or less includes 10, 9.4, 7.6, 5, 4.3, 2.9, 1.62, 0.3, etc.). Where a range of values is “up to” or “at least” a particular value, the value is included in the range and refers to a range that can be subsequently broken down into subranges as discussed above. Finally, a range includes each individual member, as will be appreciated by those skilled in the art.

[0055] As used herein, “have,” “having,” “include,” “including,” “contain,” “containing,” and the like are used in their open-ended sense, and generally mean “including, but not limited to.” It will be understood that “consisting essentially of,” “consisting of,” and the like are subsumed in “comprising,” “comprises,” and the like, and that “consisting essentially of” does not exclude the presence of additional components that do not materially affect the basic and novel characteristics of the composition, product, method, etc. The phrase “consisting of’ excludes any element not specified. As used herein, “substantially” in its reference to a given meaning indicates an extent of the given meaning that only allows for slight deviations or variations from the given meaning, such that the deviations or variations do not materially affect the overall understanding of the given meaning. As used herein, “substantially” in its reference to a given meaning indicates an extent of the given meaning that only allows for slight deviations or variations from the given meaning, such that the deviations or variations do not materially affect the overall understanding of the given meaning.

[0056] The words “preferred” and “preferably” refer to aspects of the present technology that can provide certain benefits under some circumstances. However, other aspects can also be preferred under the same or other circumstances. Furthermore, one or more preferred aspects does not imply that other aspects are not useable or exploitable and is not meant to exclude other aspects from the scope of the disclosure, including claims. DETAILED DESCRIPTION

[0057] Reference will now be made in detail to some aspects of the subject technology. While the disclosed subject technology will be described in conjunction with the enumerated claims, it will be understood that the illustrative subject matter is not intended to limit the claims to the subject technology disclosed. An aspect described in connection with a particular aspect can not necessarily be limited to that aspect and can be practiced with any other aspect.

[0058] The present disclosure relates to modified starches. In particular, the present disclosure relates to unsubstituted crosslinked waxy starches, and in particular to unsubstituted crosslinked starches. The starches of the present disclosure are cold storage stable viscosifying starches that exhibit good freeze-thaw stability. These next generation cold storage stable starches can be made without substitution (e.g., without hydroxypropyl or acetyl substitution), providing improved operational safety, reduced production costs, and environmental benefits, including less wastewater, reduced salt usage, and reduced reaction time and energy usage, thereby reducing carbon footprint and increasing sustainability of modified starches. The lack of substitution can also help avoid any supply issues of certain industrial chemicals, such as propylene oxide, which is commonly used in hydroxypropylation reactions. The cold storage stability of the unsubstituted crosslinked starches of the present disclosure can be comparable to that of hydroxypropyl-substituted crosslinked starches. The unsubstituted crosslinked starches of the present disclosure can be used as a substitute or replacement for low hydroxypropyl-substituted or medium hydroxypropyl-substituted starches or high hydroxypropyl-substituted starches.

[0059] The unsubstituted crosslinked starches of the present disclosure can be used as a substitute or replacement for acetylated starches.

[0060] Unsubstituted crosslinked starches

[0061] In one aspect, the present technology provides an unsubstituted crosslinked starch having a hot paste viscosity in the range of about 100 cP to about 1500 cP measured at pH 3 and about 5.5% solids by weight, wherein the unsubstituted crosslinked starch has a gelatinization enthalpy of about 3 J / g or less at 1 week and about 7 J / g or less at 4 weeks, and a gelatinization end temperature of less than about 75 °C.

[0062] In any aspect, the unsubstituted crosslinked starch can be prepared from waxy cassava starch. Normal cassava starch contains about 18% to about 23% amylose by weight, with the remainder being amylopectin. Waxy cassava starch contains higher levels of amylopectin and lower levels of amylose than normal cassava starch. Waxy cassava starch also contains higher levels of amylopectin and lower levels of amylose than normal cassava starch, and can be used herein to refer to cassava starch with very low amylose content. Waxy cassava starch can contain about 5% or less, about 3% or less, or about 1.5% or less amylose by weight or be a mutant that lacks the functional GBSS I gene to synthesize starch that is free of amylose. The term “GBSS I” is understood to mean any enzyme belonging to the group of granule-bound starch synthases (EC 2.4.1.21) of the subtype I.

[0063] According to an aspect of the present technology, the substituted crosslinked starch can have a chain length distribution according to the following Table 1.

[0064]

[0065] In Table 1, "Dp" refers to the degree of polymerization, and "Afp" refers to the fingerprint A chain of amylopectin. Afp chains, also known as dangling chains, are too short to participate in double helix formation during retrogradation. Fingerprint A chains (Afp) are a unique subtype of amylopectin with a Dp of 6-8, the shortest chains, and their characteristics are specific to plant sources of starch. Amylopectin molecules with a higher proportion of shorter chains are less susceptible to retrogradation.

[0066] Dp can be measured by high-performance anion exchange chromatography (HPAEC) as follows. A starch sample (2 mg) is dissolved in 90% dimethyl sulfoxide (DMSO; 50 μL), heated in a hot water bath (80°C) for 5 minutes, and then stirred for 1 hour. Warm (80°C) water (400 μL) is then added to the sample, followed by 50 μL of 0.01 M sodium acetate buffer (pH 5.5) and allowed to cool to room temperature. Isoamylase (1 μL, 465 U / mL) and 1 μL of pullulanase M1 (925 U / mL) (Megazyme) are then added, and the starch is slowly stirred overnight at room temperature (25°C) to debranch the starch. After debranching, the enzymes are inactivated by boiling for 5 minutes, the volume is adjusted to a final concentration of 1 mg / mL, and the sample is filtered through a 0.45 μm nylon filter. The filtered sample is injected into an HPAEC system equipped with a Carbopac PA-100 ion exchange column equipped with a pulsed amperometric detector. The sample was then eluted at a flow rate of 1 mL / min. The sample was eluted with the following gradient of eluent B: 15-36% B from 0-9 min; 36-45% B from 9-18 min; and 45-100% B from 18-110 min. The column was equilibrated with 15% B for 60 min between runs. Eluent A was 0.15 M NaOH (7.85 mL / 1000 mL), and eluent B was 0.15 M NaOH containing 0.50 M NaAc (7.85 mL NaOH / 41 g NaOAc for 1 L).

[0067] According to one aspect of the present technology, the unsubstituted cross-linked starch has 18 wt% or more chains with a Dp of 13 or less. The unsubstituted cross-linked starch may have 48 wt% or less chains with a Dp of 13-24. The unsubstituted cross-linked starch may have 16 wt% or more chains with a Dp of 6-12. The unsubstituted cross-linked starch may have 1.5 wt% or more chains with a Dp of 6-8.

[0068] Waxy cassava starch can be obtained from waxy cassava plants. Recessive waxy cassava mutants have been discovered in nature. Waxy cassava can be obtained through classical breeding and hybridization techniques, or through translocation, inversion, transformation, or any other genetic or chromosomal engineering method, including CRISPER / Cas9 technology. The term CRISPR as used in the art refers to Clustered Regularly Interspaced Short Palindromic Repeats. CRISPR / Cas9 refers to CRISPR associated protein 9. CRISPR / Cas9 mediated targeted mutagenesis of two genes involved in amylose biosynthesis (either targeting starch protein (PTST1) or granule-bound starch synthase (GBSS)) can reduce or eliminate amylose content in root starch. Waxy cassava starch can be extracted from the roots of cassava plants with low amylose content. Extraction can be performed by any known method, such as pulverizing the roots and extracting the starch from the pulverized roots with water. The extracted starch can be considered a native starch that has not been chemically modified.

[0069] Substitution of starch is understood to mean chemical derivatization to form ethers, esters, or half-esters, such as hydroxyalkyl ethers, acetates, phosphates, succinates (e.g., octenyl succinate), tertiary amine ethers, or quaternary amine ethers, etc., by any suitable modification technique. Typically, starch is substituted by reacting the starch with an alkylene oxide to form a hydroxyalkyl ether derivative. Etherification of starch improves the functional properties of starch in many ways. Hydroxypropylation and cross-linking are often used together to produce cross-linked, stabilized starch, which is widely used for thickening and stabilizing food applications.

[0070] According to the present disclosure, in any aspect, the starch is unsubstituted. It should be noted that cross-linking is not considered to be a substitution. In any aspect, the starch as described herein can be free of any other modification (in addition to cross-linking). The starch of the present disclosure can be characterized as an unsubstituted, cross-linked starch. For example, the starch can be free of substitution, enzymatic modification, pregelatinization, or a combination of any two or more of them. In any aspect, the starch is free of substitution, enzymatic modification, and pregelatinization. In any aspect, the starch is free of substitution and enzymatic modification, but has been pregelatinized. It has been found that the unsubstituted, cross-linked starch of the present disclosure is functionally similar to hydroxypropylated, cross-linked waxy corn starch in terms of viscosity and cold storage stability. The unsubstituted, cross-linked starch helps eliminate the typical sodium sulfate waste stream associated with the reaction of industrial propylene oxide with starch and provides energy savings and increased manufacturing capacity via a reduction in reaction time from about 22 h to less than about 8 h.

[0071] In any aspect, the starch of the present disclosure is crosslinked. The starch can be crosslinked using a crosslinking agent. Suitable crosslinking agents include epichlorohydrin, linear dicarboxylic anhydride, citric acid, acrolein, sodium trimetaphosphate, phosphorous oxychloride, adipic acid / acetate mixed anhydride, trimetaphosphate, a mixture of sodium trimetaphosphate and sodium tripolyphosphate, linear dicarboxylic anhydride, citric acid, acrolein, adipate, formaldehyde, cyanuric chloride, diisocyanate, divinyl sulfone, and combinations of any two or more thereof. In any aspect, the crosslinking agent comprises sodium trimetaphosphate (STMP), phosphorous oxychloride (POCI3), adipate, epichlorohydrin, or combinations of any two or more thereof. In any aspect, the unsubstituted crosslinked starch is suitable for use in a food product. Such starches can be crosslinked using a crosslinking agent suitable for food use. For example, a starch intended for use in a food product can be crosslinked using sodium trimetaphosphate, or phosphorous oxychloride, or a mixture of sodium trimetaphosphate and sodium tripolyphosphate. The unsubstituted crosslinked starch can be provided in any desired form. In any aspect, the unsubstituted crosslinked starch can be a non-gelatinized granular starch.

[0072] In another aspect, the unsubstituted crosslinked starch can be used to make a non-granular form of unsubstituted crosslinked starch. According to some aspects of the present technology, the unsubstituted crosslinked starch can be used to make a pregelatinized non-granular unsubstituted crosslinked starch. Pregelatinized non-granular starch can produce viscosity when dispersed in cold or warm water without the need for further heating. Pregelatinized non-granular starch is also known as pre-cooked starch, instant starch, cold water soluble starch, or cold water swelling starch. In some aspects, the unsubstituted crosslinked starch can be used to make a partially gelatinized. The starch can be partially gelatinized, for example, by spray cooking, drum drying, or extrusion. According to some aspects of the present technology, the unsubstituted crosslinked starch can be used to make a cold water swelling or instant product. The unsubstituted crosslinked starch can be used to make an agglomerated unsubstituted crosslinked starch. Crosslinked granular starch agglomerates can impart higher viscosity than non-agglomerated crosslinked starch.

[0073] Methods of making unsubstituted crosslinked starch

[0074] In any aspect, the method of making the unsubstituted crosslinked starch described herein comprises crosslinking. The method can further comprise other process steps discussed herein.

[0075] The cross-linking reaction can be performed using techniques known in the art, such as those described in U.S. Patent Nos. 2,328,537 and 2,801,242, both of which are incorporated herein in their entirety. To make unsubstituted cross-linked starch, native starch can be mixed with an aqueous solvent or water to form a slurry. The native starch can be present at a solids content of about 5% by weight or greater, about 10% by weight or greater, about 15% by weight or greater, about 20% by weight or greater, about 25% by weight or greater, about 30% by weight or greater, or about 35% by weight or greater, by weight of the slurry. The native starch can be present at a solids content of about 45% by weight or less, about 40% by weight or less, or about 35% by weight or less, by weight of the slurry. The native starch can be present at a solids content of about 20% to about 45% by weight, about 30% to about 45% by weight, or about 30% to about 40% by weight, by weight of the slurry. The pH of the slurry can be adjusted to be basic. For example, the pH of the slurry can be adjusted to a range of about 10 to about 13 or about 11 to about 12. The pH of the slurry can be adjusted with any suitable base that does not interfere with the reaction. For example, the pH of the slurry can be adjusted using sodium hydroxide (NaOH), potassium hydroxide (KOH), sodium carbonate (Na2C03), potassium carbonate (K2C03), or a combination of any two or more thereof. The slurry can also include a salt, such as sodium chloride (NaCl), sodium sulfate (Na2S04), calcium chloride (CaCl2), or a combination of any two or more thereof. The salt can be used to control the granule swelling of the starch. The inclusion of the salt can also enhance the cross-linking reaction. The salt can be included in the slurry at a concentration of 0.5% by weight or greater, about 1% by weight or greater, about 2% by weight or greater, about 3% by weight or greater, or about 4% by weight or greater. The salt can be included in the slurry at a concentration of about 10% by weight or less, about 8% by weight or less, about 7% by weight or less, or about 6% by weight or less.

[0076] The cross-linking agent can be mixed into the slurry at a suitable concentration depending on the cross-linking agent and the degree of cross-linking desired. The degree of cross-linking can be adjusted based on the viscosity desired. Starches with a higher degree of cross-linking exhibit higher viscosity than starches with a lower degree of cross-linking when compared to other similar starches. In any aspect, the amount of cross-linking agent is limited by regulatory limits on certain compounds in food products, such as phosphates. The amount of cross-linking agent in the slurry can range from 0.001 wt% to about 1.0 wt%, or 0.008 wt% to 0.1 wt%, based on dry weight. In any aspect, the amount of cross-linking agent in the slurry is 0.5 wt% or less, 0.4 wt% or less, 0.25 wt% or less, or 0.1 wt% or less, based on dry weight. The amount of cross-linking agent in the slurry can range from 0.01 wt% to 0.1 wt%. In any aspect, the amount of cross-linking agent is limited based on the amount present in the final product. For example, the amount of residual phosphate in food products and food ingredients can be limited. The amount of cross-linking agent in the final product (e.g., cross-linked starch) can be 0.4 wt% or less, 0.1 wt% or less, or 0.04 wt% or less, calculated as phosphorus. The cross-linking reaction conditions can include elevated temperatures, such as a temperature of about 25°C or higher, about 28°C or higher, or about 32°C or higher. The cross-linking temperature can be about 60°C or lower, or about 50°C or lower. The duration of the cross-linking reaction can depend on the cross-linking agent and the degree of cross-linking desired. In some cases, the duration of the cross-linking reaction can be as short as about 5-15 minutes (e.g., about 10 minutes). In some aspects, the duration can be longer than about 15 min, such as about 30 min or longer, about 60 min or longer, about 2 hours or longer, about 3 hours or longer, about 4 hours or longer, or about 5 hours or longer. The duration can be about 24 hours or shorter, about 18 hours or shorter, about 12 hours or shorter, about 10 hours or shorter, or about 8 hours or shorter. The slurry can be further held at the reaction temperature for an additional period of time (hold time). The hold time can be about 10 min or longer, about 20 min or longer, about 30 min or longer, or about 45 min or longer. The hold time can be up to about 24 hours, up to about 12 hours, up to about 6 hours, up to about 3 hours, up to about 2 hours, or up to about 1.5 hours. The hold time can be, for example, about 1 hour.

[0077] After the cross-linking reaction, the pH of the slurry can be adjusted back. For example, the pH of the slurry can be adjusted to a range of about pH 4 to about 7.5, about 4.5 to about 6.5, or about 5 to about 6. In any aspect, the pH of the slurry can be adjusted to about pH about 5.5. The pH of the slurry can be adjusted using any suitable acid, such as hydrochloric acid (HC1), phosphoric acid (H3PO4), citric acid, acetic acid, sulfuric acid, etc., or a combination of any two or more thereof.

[0078] According to any aspect of the disclosure, the chemical modification of the starch includes only cross-linking. The chemical modification of the starch can be free of substitution reactions, such as hydroxypropylation and acetylation. The unsubstituted cross-linked starch can then be dewatered and dried. Any suitable dewatering and drying method can be used. In any aspect, the unsubstituted cross-linked starch can be non-gelatinized, granular starch.

[0079] In another aspect, the unsubstituted cross-linked starch prepared according to the preparation methods described herein can be subjected to further processing to provide the unsubstituted cross-linked starch in a desired form. In any aspect, the unsubstituted cross-linked starch can be subjected to further processing to obtain a non-granular unsubstituted cross-linked starch. According to some aspects of the present technology, the unsubstituted cross-linked starch can be subjected to further processing to obtain a pregelatinized non-granular unsubstituted cross-linked starch. The pregelatinized non-granular starch can be prepared by drum drying, jet cooking, and spray drying or extrusion. The pregelatinized non-granular starch can produce viscosity when dispersed in cold or warm water without the need for further heating. The pregelatinized non-granular starch is also known as pre-cooked starch, instant starch, cold water soluble starch, or cold water swelling starch. In any aspect, the unsubstituted cross-linked starch can be subjected to further processing to obtain a partially gelatinized unsubstituted cross-linked starch. The starch can be partially gelatinized, for example, by spray cooking, drum drying, or extrusion. In any aspect, the unsubstituted cross-linked starch can be further processed to obtain a cold water swelling or instant product. The non-granular starch can be prepared by jet cooking and spray drying, roll (drum) drying, or any other thermal technique known to the skilled person. The unsubstituted cross-linked starch can be further processed to obtain an agglomerated unsubstituted cross-linked starch. Agglomerated starch is granular or instant starch that has been processed to produce granular agglomerates. A second material, such as native starch or starch derivatives (e.g., maltodextrin, dextrin, etc.), can be included to serve as a binding agent or inter-granular adhesive.

[0080] Properties of unsubstituted cross-linked starch

[0081] According to any aspect of the present technology, the unsubstituted crosslinked starch can exhibit various desirable properties. The properties described herein can apply to any unsubstituted crosslinked starch, including unsubstituted crosslinked starch. The properties of the starch can be characterized by various measurements. For example, the properties of the starch can be characterized by measuring its hot paste viscosity, retrogradation enthalpy, retrogradation end temperature, or a combination of any two or more thereof. The ability of a starch to withstand heat and shear stress is a relevant attribute for most food processing operations. The retention of strength or hot paste viscosity (HPV) reflects the shear stability of the starch granules. Hot paste viscosity is typically measured at low pH (e.g., at pH 3) or at neutral or near neutral pH (e.g., at pH about 6). Hot paste viscosity can be measured using a rapid viscosity analyzer (RVA). The RVA is a heating and cooling viscometer that measures the viscosity of a sample over a given time period while the sample is being agitated. As used herein, the hot paste viscosity of a sample refers to the viscosity at the end of the 95 °C hold time (e.g., 20 min). Retrogradation enthalpy can be measured using differential scanning calorimetry (DSC). Retrogradation enthalpy can be measured at different time intervals, such as at one week of storage and at four weeks of cold storage.

[0082] According to any aspect of the present disclosure, the unsubstituted crosslinked starch has a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) of about 90 cP or greater, about 100 cP or greater, about 200 cP or greater, about 300 cP or greater, about 400 cP or greater, about 500 cP or greater, about 600 or greater, about 700 or greater, or about 750 cP or greater. The unsubstituted crosslinked starch can have a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) of about 1500 cP or less, about 1400 cP or less, about 1300 cP or less, about 1200 cP or less, about 1100 cP or less, about 1000 cP or less, or about 900 cP or less. The unsubstituted crosslinked starch can have a hot paste viscosity in a range of about 100 cP to about 1500 cP, about 300 cP to about 1300 cP, about 400 cP to about 1200 cP, about 500 cP to about 1200 cP, about 600 cP to about 1200 cP, about 600 cP to about 1000 cP, about 400 cP to about 900 cP, or about 600 cP to about 900 cP.

[0083] According to any aspect of the disclosure, the unsubstituted crosslinked starch has a low retrogradation enthalpy and a low retrogradation end temperature. The low retrogradation end temperature and low retrogradation enthalpy indicate that the starch is less prone to retrogradation. The low retrogradation end temperature and low retrogradation enthalpy indicate that the starch has good cold storage stability. The unsubstituted crosslinked starch of the disclosure can have a retrogradation enthalpy that is low (e.g., lower than previously known starches) at both 1 week and 4 weeks of cold storage. According to any aspect of the disclosure, the unsubstituted crosslinked starch has a retrogradation enthalpy of about 3 J / g or less, about 2.8 J / g or less, about 2.5 J / g or less, about 2.4 J / g or less, about 2.3 J / g or less, about 2.2 J / g or less, about 2.1 J / g or less, or about 2.0 J / g or less measured at 1 week of cold storage. The unsubstituted crosslinked starch can have a retrogradation enthalpy of 0.7 J / g or more, 0.8 J / g or more, 0.9 J / g or more, or about 1.0 J / g or more measured at 1 week of cold storage. The unsubstituted crosslinked starch can have a retrogradation enthalpy of 0.7 J / g to about 3 J / g, 0.8 J / g to about 2.5 J / g, 0.8 J / g to about 2.4 J / g, or about 1.0 J / g to about 2.3 J / g measured at 1 week of storage. At 4 weeks of cold storage, the unsubstituted crosslinked starch can have a retrogradation enthalpy of about 7 J / g or less, about 6.7 J / g or less, about 6.5 J / g or less, about 6.3 J / g or less, about 6.2 J / g or less, about 6.1 J / g or less, or about 6.0 J / g or less. The unsubstituted crosslinked starch can have a retrogradation enthalpy of about 3.0 J / g or more, about 3.5 J / g or more, about 3.8 J / g or more, or about 4.0 J / g or more measured at 4 weeks of cold storage. The unsubstituted crosslinked starch can have a retrogradation enthalpy of about 3.0 J / g to about 7 J / g, about 3.5 J / g to about 6.7 J / g, or about 4.0 J / g to about 6.5 J / g measured at 4 weeks of cold storage. In any aspect, the unsubstituted crosslinked starch has a retrogradation enthalpy of about 3 J / g or less measured at 1 week of storage and a retrogradation enthalpy of about 7 J / g or less measured at 4 weeks of cold storage. In any aspect, the unsubstituted crosslinked starch has a retrogradation enthalpy of about 2.5 J / g or less measured at 1 week of storage and a retrogradation enthalpy of about 6.5 J / g or less measured at 4 weeks of cold storage. In any aspect, the unsubstituted crosslinked starch has a retrogradation enthalpy of about 2.0 J / g or less measured at 1 week of storage and a retrogradation enthalpy of about 6.0 J / g or less measured at 4 weeks of cold storage.

[0084] The unsubstituted crosslinked starch can have a retrogradation end temperature of about 90 °C or less, about 85 °C or less, about 80 °C or less, about 75 °C or less, or about 70 °C or less. The retrogradation end temperature can be about 50 °C or more.

[0085] The retrogradation enthalpy of the unsubstituted crosslinked starch can also be compared to the gelatinization enthalpy of the starch. The unsubstituted crosslinked starch can have a measured retrogradation enthalpy of about 25% or less, 20% or less, 15% or less, 10% or less, or 5% or less of the gelatinization enthalpy of the starch when measured after 1 week of refrigerated storage. The unsubstituted crosslinked starch can have a measured retrogradation enthalpy of about 60% or less, about 50% or less, or about 40% or less of the gelatinization enthalpy of the starch when measured after 4 weeks of refrigerated storage. The measured retrogradation enthalpy can be from 20% to 60%, from 30% to 60%, from 40% to 60%, or from 50% to 60% of the gelatinization enthalpy of the starch when measured after 4 weeks of refrigerated storage.

[0086] According to any aspect of the present application, the unsubstituted crosslinked starch has good freeze-thaw stability. Freeze-thaw cycles are considered to be a freeze-thaw combination of a composition comprising starch, such as a food product. The starch can be stable in 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 8 or more, or 10 or more freeze-thaw cycles. While there can not be an upper limit on the number of freeze-thaw cycles for starch stability, the starch can be stable in up to 12 freeze-thaw cycles. In any aspect, the starch is stable in 4 to 12 freeze-thaw cycles. Stability in freeze-thaw cycles can be assessed visually by observing whether any water separates from the product due to the storage conditions, which can include one or more freeze-thaw cycles. If no water separation is observed on the surface or when the sample is pressed, the product is considered stable under the storage conditions tested.

[0087] In any aspect, the unsubstituted crosslinked starch has a hot paste viscosity ranging from about 100 cP to about 1500 cP (measured at pH 3 and 5.5 wt% solids), a retrogradation enthalpy of about 2.3 J / g or less at 1 week and about 7 J / g or less at 4 weeks, and a retrogradation end temperature of less than about 75°C. In some cases, the unsubstituted crosslinked starch has a hot paste viscosity ranging from about 300 cP to about 1300 cP (measured at pH 3 and 5.5 wt% solids), a retrogradation enthalpy of about 2.2 J / g or less at 1 week and about 6.8 J / g or less at 4 weeks, and a retrogradation end temperature of less than about 75°C. In some cases, the unsubstituted crosslinked starch has a hot paste viscosity ranging from about 400 cP to about 900 cP (measured at pH 3 and about 5.5 wt% solids), a retrogradation enthalpy of about 2.1 J / g or less at 1 week and about 6.5 J / g or less at 4 weeks, and a retrogradation end temperature of less than about 75°C.

[0088] Unsubstituted crosslinked starches can be prepared from any suitable waxy starch source. The term "native starch" is used herein to refer to the starch source from which the unsubstituted crosslinked starch is prepared. Unsubstituted crosslinked starches can be prepared from native starches comprising about 10% or less by weight, about 5% or less by weight, about 4% or less by weight, about 3% or less by weight, about 2% or less by weight, or about 1% or less by weight amylose. The native starch can be free or substantially free of amylose. In any aspect, the unsubstituted crosslinked starch is prepared from waxy cassava, waxy rice, waxy sugar-2 corn mutant, short chain waxy potato (identified by lack of GBSS1 or non-functional GBSS1 in combination with defective or non-functional SSII and / or SSIII enzymes), waxy wheat, highly phosphorylated waxy starch, or a combination of any two or more thereof. Waxy sugar-2 is a double mutant corn plant in which the corn plant is homozygous or heterozygous recessive for the starch synthase IIa (su2) gene and homozygous or heterozygous for a mutant granule-bound starch synthase I (GBSSI) gene, wherein the mutant GBSSI gene has lower activity or no GBSSI activity. Mutant corn plants with lower GBSSI gene activity can produce less than about 10% amylose content, while no GBSSI gene activity can produce less than about 2% or zero amylose content. In any aspect, the unsubstituted crosslinked starch is prepared from waxy cassava starch. The waxy cassava starch can be obtained from naturally occurring waxy cassava plants or from waxy cassava plants developed through non-genetically modified (non-GM) or conventional breeding. The waxy cassava starch can be obtained from cassava plants modified using CRISPER / Cas9 technology. In any aspect, the unsubstituted crosslinked starch is prepared from highly phosphorylated waxy starch from potato, cassava, corn, wheat, rice, or a combination of any two or more thereof.

[0089] Use of unsubstituted crosslinked starch

[0090] According to any aspect of the present technology, unsubstituted crosslinked starches can be used in a variety of ways to provide desired properties to a product. The uses described herein can be applicable to any unsubstituted crosslinked starch, including unsubstituted crosslinked starch. The unsubstituted crosslinked starch can be formulated for use in a food product, a pet food product, or a non-food product. The unsubstituted crosslinked starch can be used in a food product, a nutritional product, a pharmaceutical product, a personal care product, a paper product, and the like. In any aspect, the unsubstituted crosslinked starch is formulated for human consumption.

[0091] Unsubstituted crosslinked starches can be used in food products to impart better cold storage stability, heat stability, thickening, increased viscosity, or a combination of any two or more thereof. For example, unsubstituted crosslinked starches can be formulated for use in dairy products, such as yogurt, sour cream, fruit toppings, dairy-based desserts, puddings, and the like. Unsubstituted crosslinked starches can be formulated for use in non-dairy yogurt, non-dairy pudding, or other non-dairy dessert. Unsubstituted crosslinked starches can be formulated for use in sauces, gravies, sauces, dips, spreads, baked good fillings, dry mixes, and the like. Unsubstituted crosslinked starches can be used in food products, such as cereal, bread, bread products, cheese, cheese products, condiments, confections, sauces, pie fillings, sauces, cheese sauces, gravies, imitation syrups, puddings, custards, yogurt, sour cream and sour cream products, pasta, beverages, milkshakes, soups, baby foods, and the like. Food products can be formulated for processing and storage conditions, such as dry retort, aseptic fill packaging, refrigeration, freezing, or a combination of any two or more thereof.

[0092] Unsubstituted crosslinked starches can be formulated for use in pet food, retort pouch food (e.g., retort pet food or any other food packaged in a retort pouch type package), or canned pet food.

[0093] Unsubstituted crosslinked starches can be used in paper products, such as paper, paperboard, linerboard, corrugated paperboard, cardboard, and the like.

[0094] Unsubstituted crosslinked starches can be used in pharmaceutical or nutraceutical products to act as a binder, disintegrant, diluent, tableting agent, dusting agent, and the like. Unsubstituted crosslinked starches can be used in personal care products, such as deodorants, antiperspirants, hair gels, gels, mousses, lotions, pomades, soaps, cleansers, shampoos, conditioners, mouthwashes, breath fresheners, toothpastes, and cosmetics, such as eye shadow, powder, foundation, blush, and the like.

[0095] According to any aspect of the disclosure, the food product comprises unsubstituted crosslinked starch. The food product can be a food product for human consumption or a pet food or animal feed. The food product can be a dairy product such as a yogurt, sour cream, fruit preparation, dairy-based dessert, pudding, etc. The food product can be a non-dairy yogurt or non-dairy pudding. The food product can be a sauce, gravy, sauce, dip, spread, baked good filling, dry mix, etc. The food product can be a pet food, retort pouch food, or canned pet food. The unsubstituted crosslinked starch can be used in the food product at any suitable concentration to achieve the desired properties or qualities affected by the presence of unsubstituted crosslinked starch. For example, the unsubstituted crosslinked starch can be included at about 1 wt% or greater, about 2 wt% or greater, about 5 wt% or greater, about 7.5 wt% or greater, about 10 wt% or greater, about 12.5 wt% or greater, about 15 wt% or greater, or about 20 wt% or greater, based on the total weight of the food product. The unsubstituted crosslinked starch can be included at about 35 wt% or less, about 30 wt% or less, about 25 wt% or less, about 20 wt% or less, about 15 wt% or less, or about 10 wt% or less, based on the total weight of the food product. In any aspect, the unsubstituted crosslinked starch comprises about 1 wt% to about 35 wt%, or about 5 wt% to about 25 wt% of the food product. In dry mixes, such as those intended for use in sauces, ready-to-eat meals, baked good fillings, etc., the starch content can be at the high end of this range, e.g., about 10 wt% to about 35 wt% or even higher, such as about 10 wt% to about 50 wt%, about 20 wt% to about 50 wt%, or about 35 wt% to about 50 wt%. The primary function of the starch in these applications is to thicken and create a paste-like texture. Lower concentrations can be used, for example, in dairy beverages, such as fermented dairy beverages, acidified dairy beverages, or neutral dairy beverages and their alternatives. The amount of starch in such products can range from 0.2 wt% to about 1 wt%. The function of the starch in these applications is to provide body and enhance mouthfeel. In yogurt, the starch can be included at a concentration of about 1 wt% to about 2.5 wt%. In these applications, the function of the starch is to bind water (providing thickening), and to provide texture and help prevent syneresis. In soups and sauces (whether or not emulsified), the starch can be included at a concentration of about 1 wt% to about 6 wt%. The function of the starch in these applications is to bind water, increase viscosity, and create texture. In fruit preparations, the starch can be included at a concentration of about 3 wt% to about 6 wt%. The function of the starch in these applications is to provide thickening, and to create a creamy texture (paste-like) and help prevent syneresis.

[0096] Exemplary Aspects

[0097] The following list provides various combinations of exemplary aspects and aspects according to the disclosure.

[0098] According to aspect 1, an unsubstituted cross-linked (e.g., cross-linked) starch having a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) ranging from about 100 cP to about 1500 cP, a gelatinization enthalpy of about 2.8 J / g or less at 1 week and about 7 J / g or less at 4 weeks, and an end temperature of gelatinized starch of less than about 75 °C.

[0099] Aspect 2 is the unsubstituted cross-linked starch according to aspect 1, wherein the unsubstituted cross-linked starch is prepared from waxy cassava, waxy rice, waxy sugar-2 corn mutant, short chain waxy potato, waxy wheat, highly phosphorylated waxy starch, or a combination of any two or more thereof.

[0100] Aspect 3 is the unsubstituted cross-linked starch according to aspect 1 or 2, wherein the starch is cross-linked.

[0101] Aspect 4 is the unsubstituted cross-linked starch according to any one of aspects 1 to 3, wherein the unsubstituted cross-linked starch comprises unsubstituted cross-linked waxy cassava starch.

[0102] Aspect 5 is the unsubstituted cross-linked starch according to any one of aspects 1 to 4, wherein the unsubstituted cross-linked modified starch is prepared from waxy cassava starch cross-linked using epichlorohydrin, linear dicarboxylic anhydride, citric acid, acrolein, phosphorous oxychloride, adipic acid / acetic acid mixed anhydride, trimetaphosphate, sodium trimetaphosphate, a mixture of sodium trimetaphosphate and sodium tripolyphosphate, linear dicarboxylic anhydride, citric acid, acrolein, adipate, formaldehyde, cyanuric chloride, diisocyanate, di-vinyl sulfone, or a combination of any two or more thereof.

[0103] Aspect 6 is the unsubstituted cross-linked starch according to any one of aspects 1 to 5, wherein the unsubstituted cross-linked starch is prepared from waxy cassava starch cross-linked using a cross-linking agent comprising sodium trimetaphosphate, phosphorous oxychloride, adipate, epichlorohydrin, or a combination of any two or more thereof.

[0104] Aspect 7 is the unsubstituted cross-linked starch according to any one of aspects 1 to 6, wherein the unsubstituted cross-linked starch is prepared from waxy cassava starch cross-linked using a cross-linking agent consisting of sodium trimetaphosphate, phosphorous oxychloride, or a combination thereof.

[0105] Aspect 8 is the unsubstituted cross-linked starch according to any one of aspects 1 to 7, wherein the hot paste viscosity (measured at pH 3 and 5.5 wt% solids) is about 200 cP or greater, about 300 cP or greater, about 400 cP or greater, about 500 cP or greater, about 600 or greater, about 700 or greater, or about 750 cP or greater.

[0106] Aspect 9 is the unsubstituted crosslinked starch of any one of aspects 1 to 8, wherein the hot paste viscosity (measured at pH 3 and 5.5 wt% solids) is about 1400 cP or less, about 1300 cP or less, about 1200 cP or less, about 1100 cP or less, about 1000 cP or less, or about 900 cP or less.

[0107] Aspect 10 is the unsubstituted crosslinked starch of any one of aspects 1 to 9, wherein the hot paste viscosity (measured at pH 3 and 5.5 wt% solids) is in the range of about 300 cP to about 1300 cP, about 400 cP to about 1200 cP, about 500 cP to about 1200 cP, about 600 cP to about 1200 cP, about 600 cP to about 1000 cP, about 400 cP to about 900 cP, or about 600 cP to about 900 cP.

[0108] Aspect 11 is the unsubstituted crosslinked starch of any one of aspects 1 to 10, wherein the retrogradation enthalpy is about 2.5 J / g or less, about 2.4 J / g or less, about 2.3 J / g or less, about 2.2 J / g or less, about 2.1 J / g or less, or about 2.0 J / g or less, measured at 1 week of refrigerated storage.

[0109] Aspect 12 is the unsubstituted crosslinked starch of any one of aspects 1 to 11, wherein the retrogradation enthalpy is about 2.3 J / g or less, measured at 1 week of refrigerated storage.

[0110] Aspect 13 is the unsubstituted crosslinked starch of any one of aspects 1 to 12, wherein the retrogradation enthalpy is about 0.7 J / g or greater, about 0.8 J / g or greater, about 0.9 J / g or greater, or about 1.0 J / g or greater, measured at 1 week of refrigerated storage.

[0111] Aspect 14 is the unsubstituted crosslinked starch of any one of aspects 1 to 13, wherein the retrogradation enthalpy is about 6.7 J / g or less, about 6.5 J / g or less, about 6.3 J / g or less, about 6.2 J / g or less, about 6.1 J / g or less, or about 6.0 J / g or less, measured at 4 weeks of refrigerated storage.

[0112] Aspect 15 is the unsubstituted crosslinked starch of any one of aspects 1 to 14, wherein the unsubstituted crosslinked starch comprises about 10 wt% or less, about 5 wt% or less, about 3 wt% or less, about 2 wt% or less, about 1.5 wt% or less amylose, or is free of amylose.

[0113] Aspect 16 is the unsubstituted crosslinked starch of any one of aspects 1 to 15, wherein the unsubstituted crosslinked starch is substantially free of amylose.

[0114] Aspect 17 is the unsubstituted crosslinked starch of any one of aspects 1 to 16, wherein the unsubstituted crosslinked starch is prepared from waxy cassava starch, wherein the waxy cassava starch is prepared from a cassava plant modified using CRISPER / Cas9 technology.

[0115] Aspect 18 is the unsubstituted crosslinked starch of any one of aspects 1 to 16, wherein the unsubstituted crosslinked starch is prepared from waxy cassava starch, wherein the waxy cassava starch is prepared from a naturally occurring waxy cassava plant or a waxy cassava plant developed through non-genetic modification or conventional breeding.

[0116] Aspect 19 is the unsubstituted crosslinked starch of any one of aspects 1 to 18, wherein the starch is not hydroxypropylated or acetylated.

[0117] Aspect 20 is the unsubstituted crosslinked starch of any one of aspects 1 to 19, wherein the starch is granular.

[0118] Aspect 21 is the unsubstituted crosslinked starch of any one of aspects 1 to 21, wherein the starch is stable in 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 8 or more, or 10 or more freeze-thaw cycles.

[0119] Aspect 22 is the unsubstituted crosslinked starch of any one of aspects 1 to 22, wherein the starch is stable in 2 to 12, 3 to 12, 4 to 12, 5 to 12, 6 to 12, 8 to 12, or 10 to 12 freeze-thaw cycles.

[0120] Aspect 23 is a pregelatinized non-granular or partially gelatinized or agglomerated product prepared using the unsubstituted crosslinked starch of any one of aspects 1 to 20.

[0121] Aspect 24 is a food product comprising the unsubstituted crosslinked starch of any one of aspects 1 to 23.

[0122] Aspect 25 is the food product of aspect 24, wherein the food product is a dairy product.

[0123] Aspect 26 is the food product of aspect 25, wherein the dairy product comprises a yogurt, a sour cream, a fruit topping, a dairy-based dessert, a pudding, or a combination of any two or more thereof.

[0124] Aspect 27 is the food product of aspect 24, wherein the food product is a non-dairy yogurt or a non-dairy pudding.

[0125] Aspect 28 is the food product of aspect 24, wherein the food product comprises a sauce, a gravy, a sauce, a dip, a spread, a baked good filling, or a dry mix.

[0126] Aspect 29 is the food product of aspect 24, wherein the food product comprises a pet food, a retort pouch food, or a canned pet food.

[0127] Aspect 30 is the food product of any one of aspects 24-29, wherein the unsubstituted crosslinked starch comprises about 1 wt% to about 35 wt%, about 5 wt% to about 25 wt, about 10 wt% to about 35 wt%, about 10 wt% to about 50 wt%, about 20 wt% to about 50 wt%, or about 35 wt% to about 50 wt% of the food product.

[0128] Aspect 31 is the food product of any one of aspects 24-30, wherein the food product comprises a dairy beverage, such as a fermented dairy beverage, an acidified dairy beverage, or a neutral dairy beverage, and the unsubstituted crosslinked starch comprises about 0.2 wt% to about 1 wt% of the food product.

[0129] Aspect 32 is the food product of any one of aspects 24-30, wherein the food product comprises a yogurt, and the unsubstituted crosslinked starch comprises about 1 wt% to about 2.5 wt% of the food product.

[0130] Aspect 33 is the food product of any one of aspects 24-30, wherein the food product comprises a soup or a sauce, and the unsubstituted crosslinked starch comprises about 1 wt% to about 6 wt% of the food product.

[0131] Aspect 34 is the food product of any one of aspects 24-30, wherein the food product comprises a fruit preparation, and the unsubstituted crosslinked starch comprises about 3 wt% to about 6 wt% of the food product.

[0132] Aspect 35 is a method of making an unsubstituted crosslinked starch, the method comprising:

[0133] mixing a native starch with water to form a slurry;

[0134] adjusting the pH of the slurry to be basic; and

[0135] mixing a crosslinking agent with the slurry to effect a crosslinking reaction,

[0136] about 1500 cP, a retrogradation enthalpy of about 2.8 J / g or less at 1 week and about 7 J / g or less at 4 weeks, and an end temperature of retrograded starch of less than about 75 °C.

[0137] Aspect 36 is the method of aspect 35, wherein the slurry comprises about 45 wt% or less, about 40 wt% or less, or about 35 wt% or less native starch dry solids, by weight of the slurry.

[0138] Aspect 37 is the method of aspect 35 or 36, wherein the slurry comprises about 20 wt% to about 45 wt%, about 30 wt% to about 45 wt%, or about 30 wt% to about 40 wt% native starch dry solids, by weight of the slurry.

[0139] Aspect 38 is the method of any of aspects 35 to 37, wherein the pH is adjusted to a range of about 10 to about 13, or about 11 to about 12.

[0140] Aspect 39 is the method of any of aspects 35 to 38, wherein the cross-linking reaction comprises a reaction temperature of about 25 °C or greater, about 28 °C or greater, or about 32 °C or greater.

[0141] Aspect 40 is the method of any of aspects 35 to 39, wherein the cross-linking reaction comprises a reaction temperature of about 60 °C or less, or about 50 °C or less.

[0142] Aspect 41 is the method of any of aspects 35 to 40, wherein the cross-linking reaction comprises a reaction temperature of about 24 °C to about 45 °C.

[0143] Aspect 42 is the method of any of aspects 35 to 41, wherein the cross-linking reaction has a duration of about 15 min or greater, about 30 min or greater, about 60 min or greater, about 2 hours or greater, about 3 hours or greater, about 4 hours or greater, or about 5 hours or greater.

[0144] Aspect 43 is the method of any of aspects 35 to 42, wherein the cross-linking reaction has a duration of about 24 hours or less, about 18 hours or less, about 12 hours or less, about 10 hours or less, or about 8 hours or less.

[0145] Aspect 44 is the method of any of aspects 35 to 43, wherein the cross-linking reaction comprises a holding time of about 10 min or greater, about 20 min or greater, about 30 min or greater, or about 45 min or greater.

[0146] Aspect 45 is the method according to any of Aspects 35-44, wherein the crosslinking reaction comprises a holding time of at most about 24 hours, at most about 12 hours, at most about 6 hours, at most about 3 hours, at most about 2 hours, or at most about 1.5 hours.

[0147] Aspect 46 is the method according to any of Aspects 35-45, wherein the crosslinking reaction comprises a duration of about 0.5 h to about 10 h and a holding time of about 10 min to about 2 hours.

[0148] Aspect 47 is the method according to any of Aspects 35-46, wherein the crosslinking agent comprises epichlorohydrin, linear dicarboxylic anhydride, citric acid, acrolein, phosphorous oxychloride, adipic acid / acetic acid mixed anhydride, trimetaphosphate, sodium trimetaphosphate, a mixture of sodium trimetaphosphate and sodium tripolyphosphate, linear dicarboxylic anhydride, citric acid, acrolein, adipate, formaldehyde, cyanuric chloride, diisocyanate, di-vinyl sulfone, or a combination of any two or more thereof.

[0149] Aspect 48 is the method according to any of Aspects 35-47, wherein the crosslinking agent comprises sodium trimetaphosphate, phosphorous oxychloride, adipate, epichlorohydrin, citric acid, or a combination of any two or more thereof.

[0150] Aspect 49 is the method according to any of Aspects 35-48, wherein the crosslinking agent consists of sodium trimetaphosphate, phosphorous oxychloride, citric acid, or a combination thereof.

[0151] Aspect 50 is the method according to any of Aspects 35-49, wherein the crosslinking agent is included in the slurry at a concentration of about 0.001 wt% to about 1.0 wt% or about 0.008 wt% to about 0.1 wt% based on dry weight.

[0152] Aspect 51 is the method according to any of Aspects 35-50, wherein the crosslinking agent is included in the slurry at a concentration of about 0.5 wt% or less, about 0.4 wt% or less, about 0.25 wt% or less, or about 0.1 wt% or less based on dry weight.

[0153] Aspect 52 is the method according to any of Aspects 35-51, wherein the slurry comprises about 0.5 wt% or more, about 1 wt% or more, about 2 wt% or more, about 3 wt% or more, or about 4 wt% or more of salt.

[0154] Aspect 53 is the method according to any of Aspects 35-52, wherein the slurry comprises about 10 wt% or less, about 8 wt% or less, about 7 wt% or less, or about 6 wt% or less of salt.

[0155] Aspect 54 is the method of any one of aspects 35-53, wherein the slurry comprises about 0.5 wt% to about 10 wt% salt.

[0156] Aspect 55 is the method of any one of aspects 52-54, wherein the salt comprises sodium hydroxide (NaOH), potassium hydroxide (KOH), sodium carbonate (Na2C03), potassium carbonate (K2C03), or a combination of any two or more thereof.

[0157] Aspect 56 is the method of any one of aspects 35-55, further comprising adjusting the pH to a range of 4 to 7.5, 4.5 to 6.5, or 5 to 6 after crosslinking.

[0158] Aspect 57 is the method of any one of aspects 35-56, further comprising dehydrating and drying the starch.

[0159] Aspect 58 is the method of any one of aspects 35-57, wherein the native starch comprises waxy cassava, waxy rice, waxy sugar-2 corn mutant, short-chain waxy potato, waxy wheat, highly phosphorylated waxy starch, or a combination of any two or more thereof.

[0160] Aspect 59 is the method of any one of aspects 35-58, wherein the native starch comprises about 10 wt% or less amylose, about 5 wt% or less amylose, about 3 wt% or less amylose, about 2 wt% or less amylose, about 1.5 wt% or less amylose, or 0 wt% amylose.

[0161] Aspect 60 is the method of any one of aspects 35-59, wherein the native starch is free or substantially free of amylose.

[0162] Aspect 61 is the method of any one of aspects 35-60, wherein the unsubstituted crosslinked starch comprises unsubstituted crosslinked waxy cassava starch.

[0163] Aspect 62 is the method of any one of aspects 35-61, wherein the unsubstituted crosslinked starch exhibits a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) of about 200 cP or greater, about 300 cP or greater, about 400 cP or greater, about 500 cP or greater, about 600 or greater, about 700 or greater, or about 750 cP or greater.

[0164] Aspect 63 is the method of any one of aspects 35-62, wherein the unsubstituted crosslinked starch exhibits a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) of about 1400 cP or less, about 1300 cP or less, about 1200 cP or less, about 1100 cP or less, about 1000 cP or less, or about 900 cP or less.

[0165] Aspect 64 is the method of any one of aspects 35-63, wherein the unsubstituted crosslinked starch exhibits a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) of about 300 cP to about 1300 cP, about 400 cP to about 1200 cP, about 500 cP to about 1200 cP, about 600 cP to about 1200 cP, about 600 cP to about 1000 cP, about 400 cP to about 900 cP, or about 600 cP to about 900 cP.

[0166] Aspect 65 is the method of any one of aspects 35-64, wherein the retrogradation enthalpy is about 2.5 J / g or less, about 2.4 J / g or less, about 2.3 J / g or less, about 2.2 J / g or less, about 2.1 J / g or less, or about 2.0 J / g or less, measured at 1 week of refrigerated storage.

[0167] Aspect 66 is the method of any one of aspects 35-65, wherein the retrogradation enthalpy is about 2.3 J / g or less, measured at 1 week of refrigerated storage.

[0168] Aspect 67 is the method of any one of aspects 35-66, wherein the retrogradation enthalpy is about 0.7 J / g or greater, about 0.8 J / g or greater, about 0.9 J / g or greater, or about 1.0 J / g or greater, measured at 1 week of refrigerated storage.

[0169] Aspect 68 is the method of any one of aspects 35-67, wherein the retrogradation enthalpy is about 6.7 J / g or less, about 6.5 J / g or less, about 6.3 J / g or less, about 6.2 J / g or less, about 6.1 J / g or less, or about 6.0 J / g or less, measured at 4 weeks of refrigerated storage.

[0170] Aspect 69 is the method of any one of aspects 35-68, wherein the method does not comprise substitution of the starch.

[0171] Aspect 70 is the method of any one of aspects 35-69, wherein the method does not comprise hydroxypropylation or acetylation.

[0172] Aspect 71 is a nutritional product comprising the unsubstituted crosslinked starch of any one of aspects 1-23.

[0173] Aspect 72 is a pharmaceutical product comprising the unsubstituted crosslinked starch according to any one of aspects 1 to 23.

[0174] Aspect 73 is a personal care product comprising the unsubstituted crosslinked starch according to any one of aspects 1 to 23.

[0175] Aspect 74 is a paper product comprising the unsubstituted crosslinked starch according to any one of aspects 1 to 23.

[0176] Aspect 75 is the unsubstituted crosslinked starch according to any one of aspects 1 to 23 having a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) ranging from about 100 cP to about 1500 cP and a retrogradation enthalpy measured upon storage at 4°C for 1 week, and a gelatinization enthalpy, wherein the retrogradation enthalpy is about 25% or less of the gelatinization enthalpy.

[0177] Aspect 76 is an unsubstituted crosslinked starch having a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) ranging from about 100 cP to about 1500 cP and a retrogradation enthalpy measured upon storage at 4°C for 1 week, and a gelatinization enthalpy, wherein the retrogradation enthalpy is about 25% or less of the gelatinization enthalpy.

[0178] Aspect 77 is the unsubstituted crosslinked starch according to aspect 76 or 77, wherein the retrogradation enthalpy is about 20% or less, 15% or less, 10% or less, or 5% or less of the gelatinization enthalpy.

[0179] Aspect 78 is the unsubstituted crosslinked starch according to any one of aspects 1 to 23 or 75 to 77, wherein the retrogradation enthalpy measured upon cold storage for 4 weeks is about 60% or less, about 50% or less, or about 40% or less of the gelatinization enthalpy of the starch.

[0180] Aspect 79 is the unsubstituted crosslinked starch according to any one of aspects 1 to 23 or 75 to 78, wherein the retrogradation enthalpy measured upon cold storage for 4 weeks is 20% to 60%, 30% to 60%, 40% to 60%, or 50% to 60% of the gelatinization enthalpy of the starch.

[0181] Example

[0182] Example 1

[0183] The pasting and retrogradation behavior of various starch samples was tested. In this study, starch retrogradation was characterized by differential scanning calorimetry (DSC). Because enthalpy (AH) reflects the melting of double helices, the enthalpy of retrograded starch reflects the degree of starch retrogradation. Samples were prepared by crosslinking native starch with different amounts of sodium trimetaphosphate (STMP) or phosphorous oxychloride (POCl3).

[0184] Slurry preparation: An alkaline slurry of native starch was prepared. In a 5 L plastic beaker, water, sodium hydroxide (NaOH), and sodium chloride (NaCl) were mixed using an overhead stirrer. The amount of water was calculated to produce a 36 wt% dry solids slurry with 1000 g of starch. The weight of NaOH was 0.52 wt% of the weight of starch. Once the NaOH was dissolved, sodium chloride (NaCl) was added at 1.5 wt% of the weight of starch. Then 1000 g of native starch was added to produce a 36 wt% dry solids slurry. The slurry was poured into a jacketed reactor with a water bath temperature set to 104 °F (40 °C) and stirred while heating.

[0185] STMP crosslinking: Once the temperature reached 105 °F (40.6 °C), sodium trimetaphosphate (STMP) was added to the slurry. The amount of STMP ranged from 0.1 wt% to 0.35 wt%. The reaction temperature was maintained for 8 h. The slurry pH was adjusted to 5.5 with dilute hydrochloric acid (HC1). The starch was dewatered through a Buchner funnel and Whatman #4 filter paper. After washing and dewatering, the starch was dried, flaked, and sieved.

[0186] POCl3 crosslinking: Phosphorous oxychloride (POCl3) was added dropwise to the slurry using a pipette. The amount of POCl3 ranged from 0.02 wt% to 0.085 wt%. The reaction temperature was maintained for 1 h. The slurry was drained into a beaker, and the pH of the slurry was adjusted to 5.5 with dilute hydrochloric acid (HC1). The starch was dewatered through a Buchner funnel and Whatman #4 filter paper. After washing and dewatering, the starch was dried.

[0187] The viscosity of the samples was measured using a Rapid Visco Analyzer (RVA) Model RVA 4800 from PerkinElmer Inc. of Waltham, MA. The samples were suspended in a pH 3 buffer or a pH 6.5 buffer at a sample content of 5.5 wt%. The total weight of the suspension was 30.0 g. Each sample was then heated to 95 °C and held at that temperature for 20 minutes at a constant 160 RPM while recording the viscosity of the sample.

[0188] Pasting parameters of starch were measured using a differential scanning calorimeter Q2000 from TA Instruments, New Castle, DE, equipped with a thermal analysis data station and data recording software (Universal Analysis 2000, also available from TA Instruments). The ratio of starch to water was 1 :3. The dispersion of starch in water was equilibrated at room temperature for 3 h prior to DSC analysis. The temperature range and heating rate of the scan were 5-105 °C and 10 °C / min, respectively. In all measurements, the thermogram curves were recorded with an empty aluminum pan as a reference. The reported transition temperatures are the onset temperature (To), the peak temperature (Tp), and the end temperature (Tc). The pasting enthalpy (AH) was estimated by integrating the area between the thermogram curve and the baseline under the peak and expressed as J / g dry starch. The pasting parameters are shown in Table 2 below.

[0189]

[0190] The retrogradation parameters of the samples were measured using a TA Instruments Q2000 differential scanning calorimeter (TA Instruments, Universal Analysis 2000) equipped with a thermal analysis data station and data recording software. The ratio of starch to water was 1 :3 and the samples were scanned from 5 °C to 105 °C at 10 °C / min. For all measurements, the thermogram curves were recorded with an empty pan as a reference. The pasted samples were cooled to room temperature (30 min) and then stored at 4 °C for 1 week, 2 weeks, 3 weeks, and 4 weeks and rescanned from 5 °C to 105 °C at 10 °C / min. In all measurements, the thermogram curves were recorded with an empty aluminum pan as a reference. The reported transition temperatures are the onset temperature (To), the peak temperature (Tp), and the end temperature (Tc). The pasting enthalpy (AH) was estimated by integrating the area between the thermogram curve and the baseline under the peak and expressed as J / g dry starch.

[0191] The RVA viscosity curves of the samples are shown in Figure 1A POCl3 crosslinked at pH 3, Figure 1B POCl3 crosslinked at pH 6.5, Figure 1C STMP crosslinked at pH 3, Figure 1B STMP crosslinked at pH 6.5, and the DSC retrogradation enthalpy results are shown in Figure 1E and Table 3 below.

[0192]

[0193] The hot paste viscosity results are shown in Table 4. The hot paste viscosity is the viscosity of the sample at the end of the hold time at 95 °C.

[0194]

[0195] Example 2

[0196] Various food samples containing starch were prepared using the starch samples prepared in Example 1. The foods included yogurt (Samples 3A, 3B, and 3C), cream dessert (Samples 4A, 4B, 4C, and 4D), baked good filling (Samples 5A and 5B), fruit topping (Samples 6A, 6B, 6C, and 6D), white sauce (Samples 7A and 7B), and pizza sauce (Samples 8A and 8B).

[0197] Yogurt samples (samples 3A, 3B and 3C)

[0198] The starch used in yogurt samples 3A, 3B, and 3C was Sample 1 (0.07 wt% POCI3 crosslinked); Sample 2 (0.35 wt% STMP crosslinked); and Sample 3 (0.085 wt% POCI3 crosslinked), respectively. Comparative samples were prepared using commercially available crosslinked, substituted waxy maize starch (hydroxypropyl distarch phosphate, available as the product number C PolarTex 06739 was obtained from Cargill, Inc. of Wayzata, MN) was used to prepare comparative samples.

[0199] Milk (UHT with 1.5 wt% fat) was mixed with 1.00 wt% skim milk powder and 1.80 wt% starch. The mixture was heated to 65°C, homogenized, and pasteurized at 95°C for 5 min. The mixture was cooled to 43°C and the lactic acid culture was mixed in. The mixture was fermented at 43°C until the pH of the mixture reached 4.6. The finished product was stored at 4°C.

[0200] After 28 days of storage at 4°C, the samples were observed to show no syneresis or change in viscosity. The texture of Samples 3A, 3B, and 3C was similar to the control.

[0201] Cream desserts (samples 4A, 4B, 4C and 4D)

[0202] The starch used in cream dessert samples 4A, 4B, 4C, and 4D was Sample 1 (0.07 wt% POCI3 crosslinked); Sample 2 (0.35 wt% STMP crosslinked); Sample 4 (0.04 wt% POCI3 crosslinked); and Sample 5 (0.2 wt% STMP crosslinked), respectively. Comparative samples were prepared using commercially available crosslinked, substituted waxy maize starch (hydroxypropyl distarch phosphate, available as the product number C PolarTex 06741 was obtained from Cargill, Inc.) was used to prepare comparative samples.

[0203] ​​Skim milk was mixed with 2.00% skim milk powder, 10.00% sucrose, 8.20% cream (35% fat content), 0.15% carrageenan, 2.00% starch, and 0.14% colorant and flavor (vanilla). The mixture was hydrated for 30 minutes and heated to 63°C. The mixture was sterilized at 135°C for 15 seconds and pre-cooled to 70-75°C. The mixture was further cooled to 8-10°C.

[0204] After storage at 4°C for 28 days, it was observed that the samples showed no syneresis. The texture of samples 4A, 4B, 4C and 4D was smooth and shiny and similar to the control. Cream dessert samples with STMP-crosslinked starch (Samples 4B and 4D) showed enhanced creaminess in sensory testing compared to PolarTex 06741 (control).

[0205] Fruit toppings (samples 5A, 5B, 5C and 5D) :

[0206] The starches used in fruit ingredient samples 5A, 5B, 5C, and 5D were sample 1 (0.07 wt% POCl3 cross-linked); sample 4 (0.04 wt% POCl3 cross-linked); sample 5 (0.2 wt% STMP cross-linked); and sample 7 (0.1 wt% STMP cross-linked). Commercially available cross-linked substituted waxy corn starch (hydroxypropyl distarch phosphate, product number C) was used. PolarTex 06741, and acetylated distarch adipate C Tex 06214, both available from Cargill) to prepare comparative samples.

[0207] Pre-blend 60g of starch with 200g of sucrose and 1g of citric acid. Blend cold fruit (500g of strawberry pulp) with 250g of cold water. Add the starch mixture to the fruit mixture, and heat the mixture to 95°C and hold at 95°C for 10 minutes. Add 0.5g of potassium sorbate. Cool the mixture to 35°C and store in the refrigerator.

[0208] After storage at 4°C for 3 months, it was observed that the samples showed no syneresis. The texture of samples 5A, 5B, 5C and 5D was similar to that of samples 5B, 5C and 5D. PolarTex 06741 and C A control made with Tex 06214 was similar.

[0209] White onion sauce (samples 6A and 6B) :

[0210] The starch used in yogurt samples 6A and 6B was sample 6 (0.02 wt% POCI3 crosslinked); and sample 7 (0.1 wt% STMP crosslinked), respectively. Comparative samples were prepared using commercially available crosslinked substituted waxy maize starch (hydroxypropyl distarch phosphate, available under product number C PolarTex 06719 from Cargill) was used to prepare comparative samples.

[0211] Dry ingredients including 9 g skimmed milk powder, 3.40 g - 4.20 g starch, 1.00 g salt and 0.50 g sodium caseinate were mixed. Water was added to the mixture. 7.50 g butter was melted at 95 °C and mixed and the water mixture was added to the melted butter. The amount of water was calculated so that the total mixture was 100 g. The mixture was cooked at 95 °C for 10 min. The sample was filled into a small pot and cooled in an ice bath.

[0212] The samples were exposed to 5 freeze-thaw cycles by freezing to -18 °C to -20 °C and thawing overnight. After 5 freeze-thaw cycles, it was observed that the samples did not show syneresis or gelling. The texture of samples 6A and 6B was similar to the control, showing little graininess and a shiny surface.

[0213] Pizza sauce (samples 7A and 7B) :

[0214] The starch used in pizza sauce samples 7A and 7B was sample 4 (0.04 wt% POCI3 crosslinked); and sample 5 (0.2 wt% STMP crosslinked), respectively. Comparative samples were prepared using commercially available crosslinked substituted waxy maize starch (hydroxypropyl distarch phosphate, available under product number C PolarTex 06741 from Cargill) was used to prepare comparative samples.

[0215] Wet ingredients including 20.00 wt% tomato sauce (28% Brix), 3.00 wt% sunflower oil and water to make the final mixture 100 wt% were mixed in a mixer (IKA mixer with tornado head). The wet mixture was transferred to a Thermomix and stirred. Dry ingredients including 5 wt% sugar, 2.60 wt% - 2.70 wt% starch and 1 wt% salt were added to the wet mixture and mixed. The mixture was heated to 95 °C with stirring and kept at this temperature for 10 min.

[0216] After 6 weeks of storage at 4 °C, it was observed that the samples did not show syneresis. The texture of samples 7A and 7B was similar to the control.

[0217] All references and publications cited herein are expressly incorporated herein by reference in their entirety unless otherwise expressly indicated. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various alternatives and / or equivalents will become apparent to those skilled in the art in view of this disclosure. It is to be understood that the herein described specific embodiments and examples are merely illustrative and are not meant to restrict the scope of the disclosure, which is defined by the claims.

Claims

1. An unsubstituted crosslinked starch having a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) in the range of about 100 cP to about 1500 cP and a retrogradation enthalpy measured at 4°C for 1 week, and a gelatinization enthalpy, wherein the retrogradation enthalpy is about 25% or less of the gelatinization enthalpy.

2. The unsubstituted crosslinked starch of claim 1, wherein the retrogradation enthalpy measured at 4°C for 1 week is 2.3 J / g or less.

3. The unsubstituted crosslinked starch of claim 1 or 2, wherein the unsubstituted crosslinked starch has a retrogradation enthalpy measured at 4°C for 4 weeks that is 50% or less of the gelatinization enthalpy.

4. The unsubstituted crosslinked starch of claim 3, wherein the retrogradation enthalpy measured at 4°C for 4 weeks is 7 J / g or less.

5. The unsubstituted crosslinked starch of any one of claims 1 to 4, wherein the unsubstituted crosslinked starch is prepared from waxy cassava, waxy rice, waxy sugar-2 corn mutant, short chain waxy potato, waxy wheat, highly phosphorylated waxy starch, or a combination of any two or more thereof.

6. The unsubstituted crosslinked starch of any one of claims 1 to 5, wherein the unsubstituted crosslinked starch is crosslinked using a crosslinking agent comprising sodium trimetaphosphate, phosphorous oxychloride, adipate, epichlorohydrin, citric acid, or a combination of any two or more thereof.

7. The unsubstituted crosslinked starch of any one of claims 1 to 6, wherein the hot paste viscosity is in the range of 400 cP to 1300 cP.

8. The unsubstituted crosslinked starch of any one of claims 1 to 7, wherein the retrogradation enthalpy is 2.0 J / g or less at 1 week.

9. The unsubstituted crosslinked starch of any one of claims 1 to 8, wherein the unsubstituted crosslinked starch comprises 5 wt% or less amylose.

10. The unsubstituted crosslinked starch of any one of claims 1 to 9, wherein the starch is not hydroxypropylated or acetylated.

11. The unsubstituted crosslinked starch of any one of claims 1 to 10, wherein the starch is granular.

12. The unsubstituted crosslinked starch of any one of claims 1 to 11, wherein the starch is stable in 4 or more freeze-thaw cycles.

13. An unsubstituted crosslinked starch having a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) in the range of about 100 cP to about 1500 cP, a retrogradation enthalpy of 2.3 J / g or less at 1 week and 7 J / g or less at 4 weeks, and a retrogradation end temperature of less than 75°C.

14. A food product comprising the unsubstituted crosslinked starch of any one of claims 1 to 13.

15. The food product of claim 14, wherein the food product is a dairy product, a non-dairy yogurt or pudding, a sauce, a gravy, a sauce, a dip, a spread, a baked good filling, a dry mix, a pet food, a retort pouch food, or a canned pet food.

16. The food product of claim 14 or 15, wherein the unsubstituted crosslinked starch comprises about 1 wt% to about 35 wt% of the food product.

17. A method of making an unsubstituted crosslinked starch, the method comprising: mixing a native starch with water to form a slurry, preferably wherein the slurry comprises about 20 wt% to about 45 wt% native starch dry solids; adjusting the pH of the slurry to basic, preferably wherein the pH is adjusted to a range of about 10 to about 13; and mixing a crosslinking agent with the slurry to effect a crosslinking reaction, preferably wherein the crosslinking reaction comprises a reaction temperature of about 24 °C to about 45 °C and a duration of about 0.5 h to about 10 h; wherein the resulting unsubstituted crosslinked starch exhibits a hot paste viscosity (measured at pH 3 and 5.5 wt% solids) in a range of about 100 cP to about 1500 cP, a retrogradation enthalpy measured at 4 °C for 1 week of storage, and a gelatinization enthalpy, and a retrogradation end temperature of less than 75 °C, wherein the retrogradation enthalpy is about 25% or less of the gelatinization enthalpy, preferably a retrogradation enthalpy of 2.3 J / g or less at 1 week and 7 J / g or less at 4 weeks.

18. The method of claim 17, wherein the unsubstituted crosslinked starch has a retrogradation enthalpy that is 50% or less of the gelatinization enthalpy measured at 4 °C for 4 weeks of storage.

19. The method of claim 17 or 18, wherein the crosslinking agent comprises sodium trimetaphosphate, phosphorus oxychloride, adipate salt, epichlorohydrin, citric acid, or a combination of any two or more thereof.

20. The method of any one of claims 17 to 19, wherein the native starch comprises waxy cassava, waxy rice, waxy sugar-2 corn mutant, short-chain waxy potato, waxy wheat, hyperphosphorylated waxy starch, or a combination of any two or more thereof.

Images