A method for rapid detection of 2'-fucosyllactose in crystals

Abstract

The invention discloses a method for quickly detecting 2'-fucosyllactose in a crystal, belongs to the technical field of 2'-fucosyllactose, and aims at the problems of poor separation degree during construction and detection of existing 2'-fucosyllactose liquid phase spectrum, unstable baseline, affecting its quantification, large fluctuation and inaccuracy during detection of crystal content. The method comprises the following steps: S1: taking a sample to be tested and a standard; S2: taking the sample and the standard in S1, diluting them with sulfuric acid, and passing through a membrane into a sample injection bottle respectively; S3: using a high performance liquid chromatograph to start sample injection detection; the invention uses an organic acid column, can completely separate a 2'-fucosyllactose base chromatographic peak under a differential detector, and has a more stable baseline than an amide bonded column chromatography, a short peak elution time, and can accurately quantify in crystal detection.

Description

Technical Field

[0001] The invention belongs to the technical field of 2'-fucosyllactose, and in particular relates to a method for rapidly detecting 2'-fucosyllactose in a crystal. Background Art

[0002] 2'-fucosyllactose is a typical representative of fucosylated human milk oligosaccharides and the most abundant functional oligosaccharide in breast milk. It has important physiological functions such as prebiotics, anti-infection, immunomodulation, and promoting brain development in infants and young children. It has now become a hot topic in the food nutrition and biopharmaceutical industries. 2'-salinalgia lactose produced by fermentation has a relatively complex composition and needs to be extracted and made into crystals for addition. Therefore, detecting the content of 2'-salinalgia lactose in the crystals is crucial for the extraction process.

[0003] The existing 2'-fucosyllactose liquid phase spectrum construction and detection have poor separation and unstable baseline, which affects its quantification. It also has large fluctuations when detecting crystal content and is not accurate enough. Therefore, we proposed a method for rapid detection of 2'-fucosyllactose in crystals. Summary of the invention

[0004] The object of the present invention is to provide a method for rapidly detecting 2'-fucosyllactose in crystals to solve the problems raised in the above background technology.

[0005] To achieve the above object, the present invention provides the following technical solution: a method for rapidly detecting 2'-fucosyllactose in crystals, comprising the following steps:

[0006] S1: Take the sample to be tested and the standard;

[0007] S2: dilute the sample and standard in S1 with sulfuric acid, and pass through the membrane into the injection bottle respectively;

[0008] S3: Use high performance liquid chromatography to start sampling and testing.

[0009] It should be noted in the scheme that the sample to be tested in step S1 is set to 10-30 mg.

[0010] It is further worth noting that the sulfuric acid content in step S2 is 0.1-0.5 mM.

[0011] It should be further explained that the column temperature in step S3 is 40°C.

[0012] As a preferred implementation, a differential detector is used as the detector in S3.

[0013] As a preferred embodiment, in step S3, the mobile phase is 0.15-0.34 mM sulfuric acid, and the flow rate is 0.4-0.7 mL / min.

[0014] As a preferred implementation, in step S3, an organic acid column is selected as the analytical column.

[0015] Compared with the prior art, the method for rapid detection of 2'-fucosyllactose in crystals provided by the present invention has at least the following beneficial effects:

[0016] The present invention uses an organic acid column, which can completely separate the 2'-fucoyl lactose chromatographic peak under a differential detector, and the baseline is more stable than that of an amide bonded column chromatography, the peak elution time is short, and accurate quantification can be achieved in crystal detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 It is a schematic diagram of the detection results of the present invention. DETAILED DESCRIPTION

[0018] The following is a detailed description of a method for rapid detection of 2'-fucoyl lactose in crystals provided by the present invention in conjunction with the accompanying drawings and specific embodiments. At the same time, it is explained here that in order to make the embodiments more detailed, the following embodiments are the best and preferred embodiments, and those skilled in the art may also adopt other alternatives to implement some known technologies; and the accompanying drawings are only for a more specific description of the embodiments, and are not intended to specifically limit the present invention.

[0019] It should be noted that the references to "one embodiment", "an embodiment", "an exemplary embodiment", "some embodiments" and the like in the specification indicate that the embodiments described may include specific features, structures or characteristics, but not every embodiment may include the specific features, structures or characteristics. In addition, when a specific feature, structure or characteristic is described in conjunction with an embodiment, it should be within the knowledge of a person skilled in the art to implement such feature, structure or characteristic in conjunction with other embodiments (whether or not explicitly described).

[0020] In general, a term can be understood, at least in part, from its use in context. For example, depending, at least in part, on the context, the term "one or more" as used herein can be used to describe any feature, structure, or characteristic in the singular sense, or can be used to describe a combination of features, structures, or characteristics in the plural sense. Additionally, the term "based on" can be understood as not necessarily intended to convey an exclusive set of factors, but can instead, depending, at least in part, on the context, allow for the presence of other factors that are not necessarily explicitly described.

[0021] It will be understood that the meanings of “on,” “over,” and “above” in the present invention should be interpreted in the broadest manner, so that “on” means not only “directly on” something, but also includes the meaning of being “on” something with intervening features or layers therebetween, and “on” or “over” means not only “on” or “above” something, but also includes the meaning of being “on” or “above” something with no intervening features or layers therebetween.

[0022] In addition, spatially relative terms such as "under," "beneath," "lower," "above," "upper," etc. may be used herein for descriptive convenience to describe the relationship of one element or feature to another element or features, as shown in the accompanying drawings. The spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the accompanying drawings. The device may be oriented in other ways, and the spatially relative descriptors used herein may be similarly interpreted accordingly.

[0023] The following examples are used to illustrate the present invention, but they cannot be used to limit the scope of protection of the present invention. The conditions in the examples can be further adjusted according to specific conditions. Simple improvements to the method of the present invention under the premise of the concept of the present invention belong to the scope of protection claimed in the present invention.

[0024] See also Figure 1 The present invention provides a method for rapidly detecting 2'-fucosyllactose in crystals, comprising the following steps:

[0025] S1: Take the sample to be tested and the standard;

[0026] S2: dilute the sample and standard in S1 with sulfuric acid, and pass through the membrane into the injection bottle respectively;

[0027] S3: Use high performance liquid chromatography to start sampling and testing.

[0028] The sample to be tested is set to any one of 10 mg, 15 mg, 20 mg, 25 mg, and 30 mg.

[0029] The sulfuric acid content in step S2 is at least one of 0.1 mM, 0.15 mM, 0.25 mM, 0.3 mM, 0.35 mM, 0.4 mM, 0.45 mM, and 0.5 mM.

[0030] The column temperature in step S3 is 40°C

[0031] Hilic organic acid column was selected as the analytical column for liquid chromatography.

[0032] The liquid chromatograph uses a differential detector, the mobile phase is any one of 0.15mM, 0.20mM, 0.25mM, 0.30mM, 0.34mM sulfuric acid, the flow rate is 0.6mL / min, and the 2'-fucosyllactose content in the crystal is calculated by reference to the standard.

[0033] Unless otherwise defined, the technical or scientific terms used in the present invention shall have the usual meanings understood by persons with ordinary skills in the field to which the present invention belongs. The words "including" or "comprising" and the like used in the present invention mean that the elements or objects appearing before the word include the elements or objects listed after the word and their equivalents, but do not exclude other elements or objects. The words "connect" or "connected" and the like are not limited to physical or mechanical connections, but may also include electrical connections, whether direct or indirect. "Up", "down", "left", "right", etc. are only used to indicate relative position relationships. When the absolute position of the object being described changes, the relative position relationship may also change accordingly.

[0034] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for rapidly detecting 2'-fucosyllactose in crystals, comprising the following steps: S1: Take the sample to be tested and the standard; S2: dilute the sample and standard in S1 with sulfuric acid, and pass through the membrane into the injection bottle respectively; S3: Use high performance liquid chromatography to start sampling and testing.

2. A method for rapid detection of 2'-fucosyllactose in crystals according to claim 1, characterized in that: In step S1, the sample to be tested is set to 10-30 mg.

3. A method for rapid detection of 2'-fucosyllactose in crystals according to claim 1, characterized in that: The sulfuric acid content in step S2 is 0.1-0.5 mM.

4. A method for rapid detection of 2'-fucosyllactose in crystals according to claim 1, characterized in that: The column temperature in step S3 is 40°C.

5. A method for rapid detection of 2'-fucosyllactose in crystals according to claim 1, characterized in that: In S3, a differential detector is used as a detector.

6. A method for rapid detection of 2'-fucosyllactose in crystals according to claim 1, characterized in that: In step S3, the mobile phase is 0.15-0.34 mM sulfuric acid, and the flow rate is 0.4-0.7 mL / min.

7. A method for rapid detection of 2'-fucosyllactose in crystals according to claim 1, characterized in that: In step S3, an organic acid column is selected as the analytical column.

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