Method for analyzing proanthocyanidin structure through combination of hydrophilic interaction and reversed-phase liquid chromatography

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: 2021-07-23
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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

Method used

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  • Method for analyzing proanthocyanidin structure through combination of hydrophilic interaction and reversed-phase liquid chromatography
  • Method for analyzing proanthocyanidin structure through combination of hydrophilic interaction and reversed-phase liquid chromatography
  • Method for analyzing proanthocyanidin structure through combination of hydrophilic interaction and reversed-phase liquid chromatography

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Embodiment 1

[0156] Embodiment 1. A method for analyzing the structure of proanthocyanidins by combining hydrophilic interaction chromatography (Hilic) and reversed-phase liquid chromatography (RPLC). The sample to be tested containing proanthocyanidins is grape seed extract, and the following steps are carried out in sequence :

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

[0158] Hilic analyzes (epi)catechin and its gallate in the sample solution to be tested, specifically 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-mentioned "1.1), Hilic analysis conditions" to detect, and then the obtained The peak area is correspondingly substituted into the formula obtained in the above "1.2)" to obtain the concentration of these 4 in the solution.

[0160] The Hilic analysis chromatogram is as follows figur...

Embodiment 2

[0183] Embodiment 2, cancel the step 3 in the above-mentioned embodiment 1 about "detecting the anthocyanidin of the acid degradation product to be analyzed at 550nm (for the acid degradation by-product)",

[0184] Therefore, the specific calculation formula of step 4) is 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 will be described in detail below.

[0187] Comparative example, traditional method:

[0188] 1) Purification of proanthocyanidins: Accurately weigh 1.0g of grape seed extract, dissolve in 4.0mL 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 eluate 1, then elute with 70% acetone aqueous solution for 3 column volumes, collect eluate 2, and eluate 2 is dried to obtain p...

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Abstract

The invention discloses a method for analyzing a proanthocyanidin structure through combination of hydrophilic interaction and reversed-phase liquid chromatography. The method comprises the following steps: analyzing a flavan-3-alcohol monomer; carrying out acid degradation on the proanthocyanidin to obtain an acid degradation product to be analyzed; analyzing the acid degradation product to be analyzed by using reversed-phase liquid chromatography (RPLC), analyzing the type and content of terminal units of proanthocyanidin according to the difference of the type and content of flavane-3-alcohol analyzed by the RPLC and Hilic, and analyzing the type and content of extension units of proanthocyanidin according to the flavane-3-alcohol-phloroglucinol addition product analyzed by the RPLC and the anthocyanidin; and calculating the average polymerization degree of the proanthocyanidin according to the mole number of the proanthocyanidin extension unit and the mole number of the terminal unit. According to the scheme, the step of directly removing flavane-3-alcohol in a sample in a traditional method (time consumption is long) is replaced, and the analysis speed of the terminal unit of the proanthocyanidin is greatly increased.

Description

technical field [0001] The invention relates to a method for quickly analyzing the structure of proanthocyanidins by combining hydrophilic interaction chromatography and reversed-phase liquid chromatography. Background technique [0002] Proanthocyanidins are a kind of polymerized polyphenols with flavan-3-ol as the structural unit, which 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)afotoin and their gallic acid esters. The structural unit is connected by C4→C6 and C4→C8 to form B-type proanthocyanidins, and the additional C2→O7 is connected 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 th...

Claims

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Application Information

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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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