Toad venom extract and preparation thereof

A technology of extract and toad venom, applied in the field of toad venom extract and its preparation, can solve the problems of complex and time-consuming gradient elution operations, and achieve the effects of improving safety and effectiveness, shortening time-consuming, and improving purification purity

Inactive Publication Date: 2011-04-20
SHANGHAI INST OF PHARMA IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, many of the conditions such as the concentration of the solvent, the sample loading method and the specific ratio of the eluent have not been disclosed, and the gradient elution operation is complicated and time-consuming
In addition, from the perspective of safety and efficacy, the total content of the three fat-soluble ingredients needs to be further improved

Method used

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  • Toad venom extract and preparation thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Take 1 gram of toad venom medicinal material, add 10ml of absolute ethanol, heat and reflux for 40min, and extract twice under the above conditions. The extract was filtered while it was hot, concentrated by rotary evaporation, and then 0.9 g of silica gel was added to mix the sample to obtain the mixed sample silica gel. Take 20 grams of silica gel soaked in petroleum ether: acetone (volume ratio 5:1) and then wet-pack the column, add sample silica gel, and perform isocratic elution with petroleum ether: acetone (volume ratio 5:1) eluent. Identification by HPLC (e.g. figure 1 As shown, the retention time is 7.184min for bufalin, 8.886min for cinobufagin, and 9.549min for bufafagenin), collect the eluate containing these three fat-soluble components, evaporate to dryness, That is to say, the toad venom extract of the present invention is obtained, wherein the mixture of three kinds of bufoligin accounts for 98.01% by weight of the whole extract.

Embodiment 2

[0021] Take 2 grams of toad venom medicinal material, add 25ml of absolute ethanol, heat and reflux for 40min, and extract twice under the above conditions. The extract was filtered while it was hot, concentrated by rotary evaporation, and then 1.8 g of silica gel was added to mix the sample to obtain the mixed sample silica gel. Take 22 grams of silica gel soaked in petroleum ether: acetone (volume ratio 5:1) and then wet-pack the column, add sample silica gel, and perform isocratic elution with petroleum ether: acetone (volume ratio 5:1) eluent. Through HPLC identification, collect the eluate containing the three fat-soluble components of bufagenin, cinobufagenin, and bufafalin, and evaporate to dryness to obtain the toad venom extract of the present invention, wherein the three kinds of bufabugenin mixtures The content is 97.94%.

Embodiment 3

[0023] Take 3 grams of toad venom medicinal material, add 35ml of absolute ethanol, heat and reflux for 40min, and extract twice under the above conditions. Filter the extract while it is hot, concentrate by rotary evaporation, add 3 grams of silica gel to mix the sample, and obtain the mixed sample silica gel. Take 22 grams of silica gel soaked in petroleum ether: acetone (volume ratio 5:1) and then wet-pack the column, add sample silica gel, and perform isocratic elution with petroleum ether: acetone (volume ratio 5:1) eluent. Through HPLC identification, collect the eluate containing the three fat-soluble components of bufagenin, cinobufagenin, and bufafalin, and evaporate to dryness to obtain the toad venom extract of the present invention, wherein the three kinds of bufabugenin mixtures The content is 99.08%.

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Abstract

The invention discloses a toad venom extract and a preparation method thereof. The preparation method comprises the following steps: (1) the toad venom medicinal material is added with absolute ethyl alcohol for reflux extraction; (2) the extracting solution obtained is condensed, separated by a silica gel column, and eluted by petroleum ether and acetone at the volume ratio of 3-10: 1 in an isocratic manner, the eluent is collected, filtered, and evaporated to be dry, thus obtaining the toad venom extract with three bufogenin mixtures. The preparation method of the invention has the advantages of being simple and convenient, being easy to operate, and being suitable for industrialized production; the content of the three bufogenin mixtures in the toad venom extract of the invention is not less than 90%.

Description

technical field [0001] The invention relates to an extract of toad venom and a preparation method thereof. The main component of the extract of toad venom is a mixture of three bufavein agglutinins, cinobufagin and bufalin. Background technique [0002] The medicinal toad is Bufo bufogargarizans Cantor or Bufo melanostictus Schneider, an animal of the family Bufoidae of the amphibian class. Toad venom is the white slurry secreted by the posterior ear glands and skin glands of Bufo bufo or Bufo bufo, which is processed and dried. Ancient and modern prescriptions widely use toads and toads to remove toxins and reduce swelling, relieve pain and kill insects, strengthen the heart and diuresis. Through modern scientific research, it has been found that the mixture of three fat-soluble components of toad bufogenin, cinobufagin and bufalin not only has obvious anti-tumor effect, but also can transform tumor cells into normal cells (induced tumor cell differentiation), induce tumo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/56A61P35/00A61K35/65
Inventor 奉建芳刘丹陈满仓刘小叶祝林
Owner SHANGHAI INST OF PHARMA IND CO LTD
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