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Extract of dogbane leaf with finger print and preparation and analytical method thereof

A technology of fingerprints and extracts, which is applied to extracts mainly containing flavonoid active substances and its preparation, an extract with fingerprints and its preparation field, which can solve the problem of long working hours, high cost and incomplete reflection Extract quality and other issues, to achieve the effect of simple operation and easy industrial production

Inactive Publication Date: 2009-02-04
周亚球 +1
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  • Summary
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AI Technical Summary

Problems solved by technology

The extract formed after collecting 40-70% of the eluent and concentrating it is the effective part, and the total flavonoid content is 50-60%. Although the content of total flavonoids in the effective parts is 50-60%, the content determination method and reference substance are not specified. As we all know, the content results obtained by different measurement methods or different reference substances of the same sample are different, such as NaNO 2 -Al(NO 3 ) 3 The content determined by the -NaOH method is significantly higher than that of AlCl 3 method, the content of the reference substance rutin is significantly higher than the hyperin reference substance, because the content of rutin in the leaves of Apocynum apocynum (except Xinjiang kenaf) is 0.07%, and the content of hyperin is 0.35%, so The content of total flavonoids in the leaves and effective parts of Apocynum apocynum can only be determined by AlCl 3 method, the reference substance can only use hyperoside
The problem with this technical solution is that although the contents of hyperin, total flavonoids, and hydrolyzed quercetin are provided, it is not clear which flavonoid components the extract consists of, and cannot fully reflect the quality of the extract. The content of total flavonoids (calculated as hyperoside) is 42.8%, which does not reach the requirement of 50% for the total flavonoid content of the five types of raw materials standards
Its preparation method is: the filtrate is subjected to polyamide column chromatography, first washed with water for 8-12 column volumes, and then eluted with different concentrations of alcohol solutions in sections, collected in sections, and concentrated. Although this method can obtain high-purity total flavonoids, but because the polyamide column needs alkali-acid regeneration after each chromatography, this method has high cost and long working hours, and is not suitable for the preparation of oral five types of raw materials

Method used

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  • Extract of dogbane leaf with finger print and preparation and analytical method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Get 100g of coarse powder of Apocynum apocynum leaves, add 800ml concentration of 75% ethanol aqueous solution, heat and reflux for extraction for 2 hours, extract 3 times in total, combine the extracts, and concentrate the extracts under reduced pressure at a temperature lower than 60°C to 1 / 5 volume, stand at room temperature for 12 hours and filter; the filtrate passes through the chromatographic column equipped with 250g AB-8 weakly polar macroporous adsorption resin at a flow rate of 1 column volume (BV) / h, and first uses water at a flow rate of 2BV / h Rinse 8 column volumes and elute with 20% ethanol to FeCl 3 If there is no dark green reaction, then use 50% ethanol to elute for 2.5 column volumes at a flow rate of 1BV / h, and collect 0.5-2.5 column volumes of the 50% ethanol eluate; depressurize the eluate below 60°C Concentrate and vacuum-dry to obtain the crude product of Apocynum apocynum leaf extract, and the total flavonoid content of the crude product is 42.0...

Embodiment 2

[0058] Get 100g of coarse powder of Apocynum apocynum leaves, add 1000ml concentration of 75% ethanol aqueous solution, heat and reflux for extraction for 2 hours, extract 3 times in total, combine the extracts, and concentrate the extracts under reduced pressure at a temperature lower than 60°C to 1 / 5 volume, stand at room temperature for 12 hours and filter; the filtrate passes through a column equipped with 250g AB-8 weakly polar macroporous adsorption resin at a flow rate of 1BV / h, and first washes 10 columns with water at a flow rate of 2BV / h volume, eluted with 20% ethanol to FeCl 3 If there is no dark green reaction, then use 50% ethanol to elute for 2.5 column volumes at a flow rate of 1BV / h, and collect 0.5-2.5 column volumes of the 50% ethanol eluate; depressurize the eluate below 60°C Concentrate and vacuum-dry to obtain the crude product of the Apocynum leaf extract, and the total flavonoid content in the crude product calculated as hyperin is 44.0%.

[0059] Refi...

Embodiment 3

[0061] Get 100g of coarse powder of Apocynum apocynum leaves, add 800ml concentration of 75% ethanol aqueous solution, heat and reflux for extraction for 2 hours, extract 3 times altogether, combine the extracts, and depressurize the extracts at a temperature lower than 60°C Concentrate to 1 / 5 volume, stand at room temperature for 12 hours and filter; the filtrate passes through the chromatographic column equipped with 250g AB-8 weakly polar macroporous adsorption resin at a flow rate of 1BV / h, and first washes 8 columns with water at a flow rate of 2BV / h Column volume, eluting with 20% ethanol to FeCl 3 If there is no dark green reaction, then use 75% ethanol to elute for 3 column volumes at a flow rate of 1BV / h, and collect 0.5-3 column volumes of the 75% ethanol eluate; depressurize the eluate below 60°C Concentrate and vacuum-dry to obtain the crude product of Apocynum apocynum leaf extract, and the total flavonoid content in the crude product of hyperin is 42.0%.

[0062...

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Abstract

A dogbane leaf extract with a fingerprint contains 50-60% of the total flavone based on hyperoside, wherein, the content of the hyperoside is 10-17%, the content of isoquercitrin is 10-16%, the contents of the isoquercitrin and kaempferol are respectively 25-35% and 3.5-4.5% after hydrolysis, the content of proanthocyanidin is 15-25%, and the extract has a definite fingerprint of the total flavone. The preparation method adopts low-polar or middle-polar macroporous absorbent resin to prepare a crude product of the extract from a concentrated solution of an extracting solution of the dogbane leaf extract, the content of the total flavone in the crude product is 40-44%, and the extract with the definite fingerprint is obtained by refining the crude product with the mixture of ethanol and ethyl acetate, and a corresponding analysis method, especially an assay method for the proanthocyanidin, is established. The extract meets the standard requirements of five oral bulk drugs, and the preparation method of the extract is convenient in operation and available in industrialization.

Description

1. Technical field [0001] The present invention relates to an active substance extracted from Chinese herbal medicine and an extraction method thereof, in particular to an extract mainly containing flavonoid active substances and a preparation method thereof. An extract with fingerprints and methods for its preparation and analysis. 2. Background technology [0002] There are three kinds of Apocynum venetum leaf resources in my country: Apocynum venetum L. (Apocynum venetum L.) is also called kenaf, Poacynum hendersonni (Hook.f.) Wodson] and Apocynum venetum L. The plant Poacynum pictum (Schrenk) Baill, collectively referred to as Apocynum pictum (Schrenk) Baill, belongs to different genera of plants, and their flavonoid components are also different, and there is no comparison between them. [0003] According to literature reports, Apocynum venetum L. contains flavonoids and their glycosides, triterpenes and sterols, lignans, coumarins, organic acids and esters, cyclic alco...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/24G01N21/31A61K127/00
Inventor 王先荣周亚球光琴杜安全陈光宗
Owner 周亚球
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