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Method for analyzing oligomeric proanthocyanidin (opc)

A technology of oligomeric proanthocyanidins and anthocyanins, which is applied in the field of oligomeric proanthocyanidins, can solve the problem of no qualitative and quantitative determination of OPC, etc.

Inactive Publication Date: 2007-06-27
SUNTORY HLDG LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] As mentioned above, there is no easy method for qualitative and quantitative determination of OPC

Method used

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  • Method for analyzing oligomeric proanthocyanidin (opc)
  • Method for analyzing oligomeric proanthocyanidin (opc)
  • Method for analyzing oligomeric proanthocyanidin (opc)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The research of embodiment 1 acid hydrolysis condition

[0060] Observe the change over time when the pine bark extract (flavangenol) is decomposed by acid. Pine bark extract (1 mg) was dissolved in 1 ml of 0.6N HCl / BuOH, and the solution was heated in a hot water bath at 90°C. Sampling was performed every 20 minutes from 20 minutes to 140 minutes. The sample was diluted 1:10 with butanol, and the visible absorption spectrum at 400-700 nm was measured. Absorption maximum occurs at 551 nm for all samples. The measurement results are shown in Table 1.

[0061] Table 1 Changes of acid decomposition of pine bark extract over time

[0062] time

[0063] The result was a slow increase in absorbance over time during acid decomposition, reaching a peak at 120 min. Based on this result, in subsequent tests, the hydrolysis period was set at 120 minutes (2 hours).

Embodiment 2

[0064] Production and determination of anthocyanins in embodiment 2 acid hydrolysis

[0065] In a glass test tube, a sample (0.5 mg) containing proanthocyanidins was dissolved in 1 ml of 0.6N HCl / butanol, and the resulting solution was left to stand in a water bath at 90° C. for 2 hours. After the reaction was completed, the absorption spectrum at 700-400 nm was measured with UV-265 (Shimadzu Corp.), and the absorbance at 551 nm was measured. Under the following conditions, use HPLC to analyze the solution after the reaction to determine anthocyanins:

[0066] Column: YMC-ODS-A312, 6mm×150mm

[0067] Mobile phase: CH 3 COOH:MeOH:H 2 O=15:17.5:67.5

[0068] Detection: A520nm (measured at 400-600nm using PDA)

[0069] The standard substances delphinidine, cyanidin and geranium pigment used for analysis were purchased from Funakoshi. The standard substance delphinium is set to flow out in 4.2 minutes, and its λ max is 535nm; cyanidine flows out in 5.5 minutes, its λ max ...

Embodiment 3

[0077] Example 3 Analysis of catechins, dimers and trimers by normal phase HPLC

[0078] Standard materials for catechins and proanthocyanidins were analyzed using normal phase HPLC under the following conditions:

[0079] The samples (0.1 mg of each sample) of (+)-catechin (Nacalai Tesque), (-)-epicatechin (Wako Pure Chemical Industries), proanthocyanidin B1 (Funakoshi) were respectively dissolved in 1 ml mobile phase, and used The above solution was filtered with a 0.45 μm filter, and then subjected to HPLC analysis under the following conditions.

[0080] The trimer was synthesized using the method described in Example 5 below.

[0081] Column: Inertsil SIL, 4.6mm×150mm

[0082] Mobile phase: hexane: MeOH: THF: HCOOH=45:40:14:1

[0083] Detection: A280nm (measured at 240-400nm using PDA)

[0084] Under these conditions, monomers ((+)-catechin and (-)-epicatechin) elute at 2.9 minutes, dimers at 3.6 minutes and trimers at 4.3 minutes.

[0085] The chromatograms of thes...

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Abstract

The present invention provides a novel method for assaying OPC contained in natural substances, foods and beverages, pharmaceuticals and / or cosmetics. The present invention is a novel method for assay of OPC which assays anthocyanidin, obtained by hydrolysis of OPC, to determine the total amount of OPC, and elucidates the proportions of the polymerization degrees of OPC by high performance liquid chromatography (HPLC) to determine the contents of the respective polymers in OPC.

Description

technical field [0001] The present invention relates to a method for determining analytes or oligomeric proanthocyanidins (the general term for n-polymer mixtures of flavan-3-ols, n≥2) contained in analytes or samples to be analyzed. Analytical samples are, for example, natural substances, food and beverages, pharmaceuticals and / or cosmetics, and the like. Background technique [0002] Proanthocyanidin (OPC) is said to be one of the active ingredients of "French Paradox", because it also contains this substance in wine (1995, Clin.Chim.Acta.235, 207-219). The known medical effects of proanthocyanidins include antioxidant activity, peripheral circulation improving activity, blood flow improving effect, liver function improving effect (2004, Japan Food Science, 403, January, 40-45) and platelet aggregation inhibitory effect (officially published Patent Publication 2003-527418). Therefore, there is a need to develop convenient methods for the qualitative and quantitative dete...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/02
CPCY10T436/142222G01N33/02G01N33/00G01N33/15G01N30/04
Inventor 福井祐子中原光一
Owner SUNTORY HLDG LTD
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