Hydrophilic interaction and reversed-phase liquid chromatography coupled to analyze the structure of proanthocyanidins

A reversed-phase liquid chromatography and proanthocyanidin technology, which is applied in the field of rapid analysis of proanthocyanidin structure, can solve the problems of time-consuming and inaccurate proanthocyanidin structure analysis results, and achieve the effect of improving analysis speed and accuracy

Active Publication Date: 2022-08-05
ZHEJIANG UNIV
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Problems solved by technology

[0005] Aiming at the problems of inaccurate and time-consuming proanthocyanidin structure analysis results, the present invention provides a method for rapidly analyzing proanthocyanidin structure by combining hydrophilic interaction chromatography (Hilic) and reversed-phase liquid chromatography (RPLC). Fast, accurate results, suitable for a variety of samples containing proanthocyanidins

Method used

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  • Hydrophilic interaction and reversed-phase liquid chromatography coupled to analyze the structure of proanthocyanidins
  • Hydrophilic interaction and reversed-phase liquid chromatography coupled to analyze the structure of proanthocyanidins
  • Hydrophilic interaction and reversed-phase liquid chromatography coupled to analyze the structure of proanthocyanidins

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Embodiment 1

[0156] Embodiment 1. A method for combined analysis of proanthocyanidin structure by hydrophilic interaction chromatography (Hilic) and reversed-phase liquid chromatography (RPLC), the test sample containing proanthocyanidin is grape seed extract, and the following steps are carried out successively :

[0157] 1), flavan-3-ol monomer analysis:

[0158] Hilic analyzes (table) catechins and their gallates in the sample solution to be tested, as follows:

[0159] Accurately weigh 5.0 mg of grape seed extract, dissolve it in 1.0 mL of 97% acetonitrile aqueous solution, and use it as the sample solution to be tested; after passing through a 0.22 μm filter membrane, use the above “1.1), Hilic analysis conditions” for detection, and then the obtained The corresponding peak areas are substituted into the formula obtained in the above "1.2)" to obtain the concentrations of these 4 in the solution.

[0160] Hilic analysis chromatogram as figure 1 shown.

[0161] 2), Proanthocyanidin...

Embodiment 2

[0183] Embodiment 2. Cancel the "detection of the anthocyanin of the acid degradation product to be analyzed at 550 nm (as the acid degradation by-product)" in step 3 in the above embodiment 1,

[0184] Therefore, the specific calculation formula of step 4) is also correspondingly changed as follows:

[0185]

[0186] In order to compare the method of the present invention with the traditional proanthocyanidin structure analysis method, the traditional method is described in detail below.

[0187] Comparative example, traditional method:

[0188] 1) Purification of proanthocyanidins: Accurately weigh 1.0 g of grape seed extract, dissolve it in 4.0 mL of 50% methanol aqueous solution, load the sample on a Sephadex LH20 column equilibrated with 50% methanol aqueous solution, and elute 5 columns with 50% methanol aqueous solution volume, collect eluent 1, then elute with 70% acetone aqueous solution for 3 column volumes, collect eluent 2, and dry eluent 2 to obtain purified g...

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Abstract

The invention discloses a method for analyzing the structure of proanthocyanidins by combining hydrophilic interaction and reversed-phase liquid chromatography, comprising the following steps: analyzing flavan-3-alcohol monomers; Acid degradation product; the acid degradation product to be analyzed is analyzed by reversed-phase liquid chromatography (RPLC), and the type and content of proanthocyanidin end units are analyzed according to the difference in the type and content of flavan-3-alcohol analyzed by RPLC and Hilic, according to RPLC The analyzed flavan-3-alcohol-phloroglucinol addition products and anthocyanins were analyzed for the type and content of proanthocyanidin extension units; then the average degree of polymerization of proanthocyanidins was calculated according to the number of moles of proanthocyanidin extension units and terminal units. This solution replaces the step of directly removing flavan-3-alcohols in the sample (which takes a long time) in the traditional method, and greatly improves the analysis speed of the terminal units of proanthocyanidins.

Description

technical field [0001] The invention relates to a method for rapidly analyzing the structure of proanthocyanidins by combining hydrophilic interaction chromatography and reversed-phase liquid chromatography. Background technique [0002] Proanthocyanidins are a class of polymeric polyphenols with flavan-3-ols as structural units, and exhibit different structures due to different unit types, degrees of polymerization and connection methods. The structural units of proanthocyanidins include (epi)catechin, (epi)gallocatechin, (epi) afomonin and their gallates. The structural units are connected by C4→C6 and C4→C8 to form B-type proanthocyanidins, and additional C2→O7 to form A-type proanthocyanidins. A large number of studies have shown that proanthocyanidins have a variety of active functions such as good antioxidant properties, prevention of cardiovascular and cerebrovascular diseases, and weight loss, and these functions are closely related to their structure, especially th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/89
CPCG01N30/89
Inventor 潘海波叶兴乾陈士国程焕陈健乐徐新雷钱子琪王艺
Owner ZHEJIANG UNIV
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