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A method for separating flavonoids in custard apple leaves

A technology of custard apple leaves and flavonoids, which is applied in the field of high-purity flavonoid monomers, can solve the problems of low preparation efficiency of flavonoid compound monomers, low sample purity, difficult preparation volume, etc., and achieve good anti-oxidation and hypoglycemia in vitro Activity, simple production process, saving the effect of preparation process

Active Publication Date: 2021-03-30
济南市疾病预防控制中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The inventors have found that the traditional separation methods for flavonoids in custard apple leaves are reverse phase chromatography, silica gel column chromatography and Sephadex LH-20, etc., which are time-consuming and laborious, pollute the environment, have low sample purity, have limitations, and repeat column chromatography It has an irreversible adsorption effect on the sample, and the preparation efficiency of flavonoid monomers obtained by separation is low, and the cost is high, so it is difficult to develop into a separation technology with a large amount of preparation

Method used

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  • A method for separating flavonoids in custard apple leaves
  • A method for separating flavonoids in custard apple leaves
  • A method for separating flavonoids in custard apple leaves

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] 1.1 Sample extraction

[0068] The dried custard apple leaves (2kg) were completely crushed into powder, extracted twice with 5L of 95% ethanol under reflux, each time for 2h, and then the extract was filtered with a vacuum filter. Combine all extracts and concentrate until there is no alcohol smell, then use equal proportion of petroleum ether to extract, degrease, and then use equal proportion of n-butanol to extract the water phase twice, n-butanol extract is concentrated under reduced pressure, dried to obtain the concentrate 62g, that is, a sample of crude flavonoids. The primary crude flavonoid sample is subjected to secondary degreasing, the secondary degreasing solvent is petroleum ether / ethyl acetate / methanol / water (5:5:2:8, v / v), the lower phase is concentrated under reduced pressure, dried, and the total flavonoid sample is obtained 17.6g.

[0069] 1.2 Separation and purification of flavonoid monomers using on-line collection and circulating countercurrent ...

Embodiment 2

[0086] 1. Sample Extraction

[0087] The dried custard apple leaves (2kg) were completely crushed into powder, extracted twice with 5L of 95% ethanol under reflux, each time for 2h, and then the extract was filtered with a vacuum filter. Combine all extracts and concentrate until there is no alcohol smell, then use equal proportion of petroleum ether to extract, degrease, and then use equal proportion of n-butanol to extract the water phase twice, n-butanol extract is concentrated under reduced pressure, dried to obtain the concentrate And carry out secondary degreasing, the secondary degreasing solvent system is petroleum ether / ethyl acetate / methanol / water (5:5:2:8, v / v), the lower phase is concentrated under reduced pressure, dried, and obtains the total sample of flavonoids .

[0088] 2.1 Separation of total flavonoids from custard apple leaves by linear gradient high-speed countercurrent chromatography

[0089] The specific operation steps are: adopt a semi-preparative h...

Embodiment 3

[0102] 1. Method

[0103] 1. Evaluation method of antioxidant activity

[0104] DPPH (2.5 mg) was dissolved in 100 mL of ethanol to prepare a standard solution with a concentration of 25 μg / mL. Serial dilutions were used to provide ethanol standard solutions at concentrations of 0, 5, 10, 15, 20 and 25 μg / mL. The absorbance values ​​of the six solutions were measured at 517 nm by UV spectrophotometry to prepare a standard curve. DPPH (2.0 mg) was dissolved in 100 mL of ethanol to prepare a standard solution with a concentration of 20 μg / mL. Solutions containing different concentrations of L-ascorbic acid (positive control), crude extracts and monomeric compounds were prepared as test samples. Add 3mL of the standard solution into a 10mL colorimetric tube together with 2mL of the solution from the sample set. For the control group, add 3mL of ethanol and 2mL of sample solution into a 10mL colorimetric tube, and use 3mL of standard solution containing 2mL of ethanol as a bla...

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Abstract

The invention relates to the field of extraction of natural compounds, in particular to a method for separating flavonoid components in annona squamosa leaves. An ethanol extract of the annona squamosa leaves is subjected to dealcoholization, petroleum ether degreasing and n-butyl alcohol extraction to obtain a crude flavonoid extract, and the crude flavonoid extract is distributed by a two-phasesolvent to obtain a total flavonoid sample. By means of high-speed countercurrent chromatography, a secondary flavonoid extract is separated and purified to obtain quercetin-3-O-beta-D-glucoside, kaempferol-3-O-robinobioside, kaempferol-3-O-rutinoside, quercetin-3-O-robinobioside and rutin, and the purity is 95% or above. The invention further provides a method for separating the flavonoid components through linear elution. High-purity monomer can be obtained, and the method has good popularization significance.

Description

technical field [0001] The disclosure relates to the field of extraction of natural compounds, in particular to a method for extracting five high-purity flavonoid monomers from custard apple leaves based on high-speed countercurrent chromatography. Background technique [0002] Annonasquamosa (scientific name: Annonasquamosa Linn.) Annonasquamosa Linn. Annonasquamosa is a small deciduous tree in the family Annonasquamosa; the bark is thin, off-white, and multi-branched. It is cultivated in China's Zhejiang, Taiwan, Fujian, Guangdong, Guangxi, Hainan and Yunnan provinces. Native to tropical America; now cultivated in tropical regions of the world. The fruit is eaten, and its shape resembles a lychee, so it is called "Annan apple", which is a famous fruit in tropical areas. The leaves of custard apple are a class of medicinal and edible herbal medicines. They are often made into health teas in my country. They are bitter, astringent, and slightly cold in nature. Return larg...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07H17/07C07H1/08A61P39/06A61P9/00A61P11/14
CPCA61P9/00A61P11/14A61P39/06C07H1/08C07H17/07
Inventor 白爱英刘仲刘岚铮王岱杰李景超宋祥云
Owner 济南市疾病预防控制中心
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