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Method used for separating flavonoid components, and applications thereof

A technology for flavonoids and structure separation, which is applied in the field of analytical chemistry and can solve the problems of separation, time-consuming and high detection cost of astragaloside and quercitrin.

Active Publication Date: 2018-07-13
GUANGZHOU BAIYUSN HUTCHISON WHAMPOA CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reason may be that the chromatographic conditions used in the above study failed to separate ascarin and quercetin
Moreover, these methods are all based on the external standard method, requiring multiple corresponding standard products to establish standard curves for each component, which is cumbersome, time-consuming, and costly to detect, and is not suitable for the requirements of industrial production quality control.

Method used

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  • Method used for separating flavonoid components, and applications thereof
  • Method used for separating flavonoid components, and applications thereof
  • Method used for separating flavonoid components, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1 Study on Separation of Flavonoids by High Performance Liquid Chromatography

[0093] 1. Preparation of mixed reference solution

[0094] Accurately weigh the appropriate amount of reference substances of hyperoside, isoquercitrin, astragalin, quercitrin, myricetin, quercetin, and kaempferol respectively, place them in 25mL volumetric flasks, add methanol to volume to the scale line, shake well, and formulate the mass concentrations of hyperoside 0.8116mg / mL, isoquercitrin 0.7092mg / mL, astragalin 0.7668mg / mL, quercitrin 0.7820mg / mL, myricetin 0.8332mg / mL, quercetin 0.7592mg / mL, kaempferol 0.7980mg / mL reference substance stock solution.

[0095] Take 5mL of each of the above-mentioned reference substance stock solutions, place them in the same 100mL volumetric flask, add methanol to the volume, shake well, and prepare A mixed reference solution of 0.03834 mg / mL baexin, 0.03910 mg / mL quercitrin, 0.04166 mg / mL myricetin, 0.03796 mg / mL quercetin, and 0.03990 mg...

Embodiment 2

[0105] Example 2 The method established in Example 1 detects flavonoids in the persimmon leaf extract

[0106] 1. Preparation of mixed reference solution

[0107] Prepare the mixed reference substance solution according to the same method under the item "1." of Example 1.

[0108] 2. Preparation of the test solution:

[0109] Take persimmon leaf extract, take about 0.1g, weigh it accurately, put it into a stoppered Erlenmeyer flask, add 20mL of methanol precisely, seal it tightly, weigh it, ultrasonically treat it (power 250W, frequency 45kHz) for 30min, take it out, and let it cool , and then weighed, make up the lost weight with methanol, shake well, filter with a 0.45 μm microporous membrane, and take the subsequent filtrate to obtain the test solution of persimmon leaf extract.

[0110] 3. Preparation of sample for mixing and adding samples

[0111] Take the persimmon leaf extract, take 1 / 2 (about 0.05g) of the sample amount of the persimmon leaf extract test sample, ...

Embodiment 3

[0126] Example 3 Establishment of a method for determining the content of flavonoids in persimmon leaf extract based on one test and multiple evaluation

[0127] 1. Preparation of mixed reference solution

[0128] Accurately weigh the appropriate amount of reference substances of hyperoside, isoquercitrin, quercitrin, myricetin, quercetin, kaempferol and astragalin respectively, place them in a 25mL volumetric flask, add methanol to the volume Scale line, shake well, and formulate the mass concentration as hyperoside 0.8116mg / mL, isoquercitrin 0.7092 mg / mL, quercetin 0.7820mg / mL, myricetin 0.8332mg / mL, quercetin 0.7592mg / mL, the mixed reference stock solution of kaempferol 0.7980mg / mL and ascarin 0.7668mg / mL.

[0129] Accurately draw 10.0, 5.0, 3.0, 2.0, 1.0, 0.5, 0.1mL of the above-mentioned mixed reference substance stock solutions respectively, place them in a 25mL volumetric flask, dilute with methanol to the mark, and shake well to prepare a series of mixed reference su...

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Abstract

The invention relates to a method used for separating flavonoid components of the similar structures. The method is invented based on high performance liquid chromatography; the flavonoid components of the similar structures at least comprise astragalin and quercitrin; the chromatography conditions are as following: octadecylsilane chemically bonded silica is taken as a fixing phase, acetonitrileis taken as a mobile phase A, 0.05 to 0.5% (percent by volume) of a phosphoric acid aqueous solution is taken as a mobile phase B, and gradient elution adopted, 0 to 40min of 7%-25% A, and the balanceB; or 40 to 60min of 25%-50% A, and the balance B. The method is capable of realizing separation of astragalin and quercitrin for the first time. The invention also provides applications of method indetecting flavonoid components in persimmon leaf extract product, and especially applications in measuring of the content of flavonoid components in persimmon leaf extract product or preparations ofpersimmon leaf extract product based on quantitative analysis of multi-components by single-marker.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and in particular relates to a method for separating flavonoid components with similar structures based on high performance liquid chromatography and an application thereof. Background technique [0002] Flavonoids are a series of compounds with 2-phenylchromone as the core, which have a wide range of physiological activities, such as anti-oxidation, anti-inflammation, anti-allergy, repair of damaged DNA and so on. However, because of its relatively simple core structure and limited types of substituents, mainly hydroxyl, methoxy, sugar, etc., it is difficult to separate flavonoids with similar structures even with the help of a high-efficiency liquid phase system. For example hyperoside (structural formula shown in 1), isoquercitrin (structural formula shown in 2), quercitrin (structural formula shown in 3), astragalin (structural formula shown in 4), myricetin (structural formula as shown i...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N30/06G01N30/02G01N3/14C07H17/07C07H1/06
CPCC07H1/06C07H17/07G01N3/14G01N30/02G01N30/06G01N30/88G01N2030/027G01N2030/062G01N2030/146G01N2030/884
Inventor 郭海彪苏诗韵李楚源王德勤李淑如
Owner GUANGZHOU BAIYUSN HUTCHISON WHAMPOA CHINESE MEDICINE
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