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Method for determining content of organic acid in ginkgo leaf extract

A determination method and extract technology are applied in the field of organic acid content determination in Ginkgo biloba extract, which can solve the problems of large polar span of proanthocyanidins and inability to completely remove interference, improve separation effect, facilitate balance, and accurately quantify organic acids. acid effect

Active Publication Date: 2017-05-31
云南康恩贝植物研究院有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Macroporous resin purification method is a commonly used method for purifying proanthocyanidins, but the polarity span of proanthocyanidins is very large, monomer components, such as catechins, are slightly soluble in cold water, while high-polymer proanthocyanidins are easily soluble in cold water, and proanthocyanidins The higher the degree of polymerization, the greater the polarity, so the macroporous resin can be used to adsorb proanthocyanidins, but the high-polymer proanthocyanidins and organic acid components with higher polarity can be eluted with water or low-concentration ethanol aqueous solution, but still cannot be completely absorbed. Remove distractions

Method used

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  • Method for determining content of organic acid in ginkgo leaf extract
  • Method for determining content of organic acid in ginkgo leaf extract
  • Method for determining content of organic acid in ginkgo leaf extract

Examples

Experimental program
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Effect test

Embodiment 1

[0031] The impact of different purification methods of embodiment 1 on the determination of organic acids

[0032] Purification method 1: 100mg of Ginkgo biloba extract, 10ml of aqueous solution containing 1g / L bovine serum albumin, ultrasonically extracted at 50°C for 60min, filtered; the filtrate was adsorbed with HPD5000 macroporous resin, washed with water, eluted with 10% ethanol, collected and adsorbed The residue, the parts washed with water and the parts eluted with 10% ethanol were concentrated to dryness, and the residue was dissolved with 0.1% phosphoric acid, and the volume was adjusted to 10ml.

[0033] Purification method 2: extract 100 mg of ginkgo leaf extract with 10 ml of an aqueous solution with a pH value of 13, adjust the pH value of the extract to 1 with concentrated hydrochloric acid, and then extract twice with 100 ml of ethyl acetate. The two ethyl acetate solutions were combined, concentrated to dryness, and the residue was dissolved with 0.1% phospho...

Embodiment 2

[0037] Qualitative and quantitative of organic acid components in the Ginkgo biloba extract of embodiment 2

[0038] Take 100mg of ginkgo biloba extract, use 10ml of aqueous solution containing 1.0g / L bovine serum albumin, ultrasonically extract at 50°C for 1h, and filter; the filtrate is concentrated and dried to obtain an organic acid extract; The aqueous solution was dissolved and the volume was adjusted to 10ml, shaken up, filtered, and the subsequent filtrate was taken; the subsequent filtrate was determined according to the chromatographic conditions of Example 1.

[0039] The above method was validated by methodology, including instrument precision, repeatability, intermediate precision, linearity, sample recovery, and durability.

[0040] It was found that Ginkgo biloba extract contains shikimic acid, gallic acid, protocatechuic acid, 6-HKA, and p-hydroxybenzoic acid, but does not contain ferulic acid, chlorogenic acid, isochlorogenic acid A, and isochlorogenic acid B....

Embodiment 3

[0055] The investigation of embodiment 3 bovine serum albumin consumption

[0056] Take 100mg of ginkgo biloba extract, use 10ml of aqueous solution containing 0, 0.1, 0.3, 1.0, 3.0, 10g / L bovine serum albumin respectively, ultrasonically extract at 50°C for 1h, and filter; the filtrate is concentrated and dried to obtain an organic acid extract; The acid extract was dissolved with an aqueous solution containing 0.1% phosphoric acid and the volume was adjusted to 10ml, shaken up, filtered, and the subsequent filtrate was taken; the subsequent filtrate was determined according to the chromatographic conditions of Example 1.

[0057] see attached results figure 2 , attached Figure 5 And attached Figure 6 . attached figure 2 It is the chromatogram of the sample pretreated with 1.0g / L bovine serum albumin, and the 5 main peaks are in good shape without interference from other peaks. The chromatogram of the sample pretreated with 3.0 or 10g / L bovine serum albumin is attache...

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Abstract

The invention discloses a method for determining the content of organic acid in a ginkgo leaf extract. The method comprises the following steps: pretreatment of a ginkgo leaf extract sample; preparation of a test article solution, high performance liquid chromatograph analysis and the like. The method is characterized in that ultrasonic extraction is carried out by using an aqueous solution containing bovine serum albumin in a pretreatment process of the ginkgo leaf extract sample, so that the interference of procyanidine is removed; a sample is prepared by using an aqueous solution containing 0.1 percent of phosphoric acid in a preparation process of the testing article solution, so that the interference of part of impure peaks is removed, and the chromatographic behavior and the separation effect are greatly improved. After the method is adopted for analyzing, an organic acid pattern of the ginkgo leaf extract can be obtained, and the contents of five organic acid components such as shikimic acid, gallic acid, protocatechuic acid, 6-hydroxykynurenic acid and p-hydroxybenzonic acid can be simultaneously quantitated.

Description

technical field [0001] The invention relates to a method for determining the content of organic acids in ginkgo biloba extract. Background technique [0002] Ginkgo biloba is the dried leaf of Ginkgo biloba (Ginkgobiloba L.), a plant of the family Ginkgoaceae. Ginkgo biloba extract is clinically used to treat cardiovascular and cerebrovascular diseases and nervous system diseases. The components of Ginkgo biloba extract are complex, and the current quality control index has only two types of components: flavonol glycosides and lactones, the total content of which is 30%. Another 70% of the ingredients are not included in the quality standards. It has been reported in the literature that in addition to flavonol glycosides and lactones, Ginkgo biloba extract also contains proanthocyanidins, organic acids, biflavones, isoflavones and other components. Therefore, in order to gradually increase the measurable components in Ginkgo biloba extract, gradually improve and improve t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88
Inventor 乔洪翔刘华春任红蒋春黄海波陈君陈岑孔琼荣王莲王如伟
Owner 云南康恩贝植物研究院有限公司
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