Method for procyanidin analysis

A technology for proanthocyanidins and catechins, which can be used in analytical materials, biological material analysis, material separation, etc., and can solve the problems of impossible removal by quantitative analysis, limited compounds, and difficult quantitative methods.

Inactive Publication Date: 2007-01-10
SUNTORY HLDG LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in any analysis method, proanthocyanidins mixed with other ingredients in tea and nutritional supplements are difficult to separate from impurity peaks, so it is difficult to say that it is an accurate quantitative method
Moreover, there are many stereoisomers of proanthocyanidins due to the stereoisomerization of flavan-3-ols, and the compounds that can be obtained as standard substances are limited, so it is impossible to remove some known compounds by quantitative analysis.

Method used

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  • Method for procyanidin analysis
  • Method for procyanidin analysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] [Example 1] Discussion of pretreatment conditions

[0038] 5 ml of an aqueous solution containing 20 ppm each of proanthocyanidin B1 (PB1) and caffeine was loaded on a chromatographic column swelled with water to make Sephadex LH-20 (Amersham Biosciences, Inc., Japan) with a dry weight of 0.25 g, and washed with 2 ml of water , eluted with 2 ml each of 20%, 40%, 60%, and 80% EtOH. After each eluted fraction was concentrated under reduced pressure concentration, it was packed in a 2ml volumetric flask, and each eluted fraction was used for HPLC analysis (analysis conditions were recorded in Example 3) to quantify caffeine and PB1.

[0039] extract

[0040] From the results, it can be seen that caffeine is almost absorbed by water (H 2 O) elution, and PB1 is eluted by equal to or greater than 60% EtOH, and the two can be separated.

Embodiment 2

[0041] [Embodiment 2] Sephadex LH-20 pretreatment of tea products

[0042] 5 ml of tea products containing Flavangenol (proanthocyanidins derived from pine bark) were loaded on a chromatographic column swelled with Sephadex LH-20 (Amersham Biosciences Co., Ltd., Japan) with a dry weight of 0.25 g with water, washed with 2 ml of water, Elute with 35% EtOH 2ml, 70% EtOH 4ml. After the 70% EtOH eluted fractions were concentrated under reduced pressure, they were packed into a 2ml volumetric flask, and each eluted fraction was analyzed by HPLC (the analysis conditions were described in Example 3) to quantify caffeine and PB1. Concentrations were converted to those corresponding to the original volume (5 ml).

[0043] extract

caffeine concentration

PB1 concentration

h 2 O non-adsorbed

h 2 o wash

35%EtOH

70%EtOH

100ppm

9ppm

0.2ppm

0ppm

0ppm

0ppm

0.08ppm

2.7ppm

[0044] Wash with water to almost...

Embodiment 3

[0046] [Example 3] Separation of proanthocyanidins and flavan-3-alcohols with HPLC

[0047] Under the following conditions, HPLC was used to analyze the water, 35% or 70% EtOH elution fraction, and the sample solution after pretreatment to remove impurities such as caffeine, which were separated in the Sephadex LH-20 treatment.

[0048] The analysis conditions are that the chromatographic column adopts Capcellpak C-18 AQ (6mm×150mm, Japan Shiseido Co., Ltd.), the A solution is 0.05% TFA / water, and the B solution is 0.05% TFA / 90% CH 3 CN / water, flow rate 1.2ml / min, carry out gradient elution of B10%→B10% (10 minutes) B10%→B35% (10 minutes). The column temperature is 40°C, and the area value of A225nm is used for quantification. HPLC uses Shimadzu LC-2010HT (Shimadzu Corporation, Japan).

[0049] Under this condition, proanthocyanidin B1 scored 10.7 points, proanthocyanidin B3 scored 12.6 points, catechin scored 14.0 points, epicatechin scored 17.6 points, gallocatechin scored...

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Abstract

The present invention relates to a novel method for quantifying procyanidin and flavan-3-ols contained in natural products, foods and drinks, pharmaceuticals and / or cosmetics. The present invention provides a method for quantifying procyanidin (a generic name for a mixture of catechin n-mer; n >= 1) contained in test samples such as natural products, foods and drinks, pharmaceuticals and / or cosmetics, which comprises a) pretreating a test sample using a column to remove contaminants which affect the analysis of procyanidin and flavan-3-ol; and b) effecting high performance liquid chromatography (HPLC) to separate and quantify procyanidin and flavan-3-ols in the solution treated to remove contaminants during the above pretreatment step a).

Description

technical field [0001] The present invention relates to a method for quantifying proanthocyanidins and flavan-3-ols contained in natural substances, food and beverages, pharmaceuticals and / or cosmetics. Background technique [0002] As far as the efficacy of proanthocyanidin (OPC) is concerned, because it also exists in wine, it is considered to be one of the active ingredients of "French Paradox" (1995, Clin.Chim.Acta., 235, 207-219), which are known to have antioxidant effects, peripheral circulation improvement effects, blood fluidity improvement effects, liver function improvement effects (2004, Japan Food Science, 403, January issue, 40-45), platelet aggregation Inhibition effect (Special Table 2003-527418), etc. [0003] The currently known analytical methods of proanthocyanidins are: reverse phase HPLC (2003, Biosci.Biotechnol.Biochem., 67 (5), 1140- 1142), LC-MS gradient elution normal phase HPLC (2003, J. Agric. Food Chem., 51, 7513-7521). However, in any analysi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/00G01N30/89G01N30/14B01D15/08
CPCG01N2030/8813G01N33/14Y10T436/142222
Inventor 福井祐子
Owner SUNTORY HLDG LTD
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