Extraction and purification process of anticoagulant peptide in centipedes

Abstract

The invention discloses an extraction and purification process of anticoagulant peptide in centipedes. The process comprises the following steps: freezing fresh and live adult centipedes by using liquid nitrogen and grinding the frozen fresh and live adult centipedes into powder; adding an ammonium sulfate aqueous solution and a protease K aqueous solution into the powder and treating for 5 to 15 minutes; then putting into boiling water and boiling for 2 to 8 minutes; then putting on ice and cooling, and evenly shaking and then centrifugally fetching supernatant liquid; adding absolute ethyl alcohol into the supernatant liquid, evenly mixing and then centrifugally fetching an upper layer of protein liquid; adding a sodium sulfate aqueous solution of which the volume is 0.1 to 0.5 time of that of the upper layer of protein liquid and the mass percentage is 10 to 30 percent to obtain a mixture; evenly mixing the mixture and then adding the absolute ethyl alcohol of which the volume is 1 to 2 times of that of the mixture; evenly mixing the mixture and then centrifuging the mixture for 15 to 25 minutes at speed of 12000 to 14000r / min, and collecting an upper layer of solution; the fifth step: freeze-drying the upper layer of solution so as to be made into freeze-dried powder. According to the process disclosed by the invention, anticoagulant peptide in the centipedes can be quickly and effectively extracted.

Description

technical field [0001] The invention relates to an extraction and purification process of active ingredients, in particular to an extraction and purification process of centipede anticoagulant peptide, and belongs to the technical field of deep processing of pharmaceutical raw materials. Background technique [0002] Centipede, also known as Tianlong and Millennium Bug, is an animal of the Centipede family of the Arthropoda Phylum Chiliopoda Plastic Order. Its medicinal use was first recorded in "Shen Nong’s Materia Medica", which has a medicinal history of more than two thousand years; "Compendium of Materia Medica" is contained in The forty-second volume of the Ministry of Insects is a traditional Chinese medicinal material for animals; it has been included in the "Pharmacopia of the People's Republic of China" in modern times. Its dry worm body is pungent in nature, warm in taste, poisonous, and belongs to the Liver Meridian. Wherever qi and blood condense, it can be ope...

AI Technical Summary

Problems solved by technology

[0004] Centipede toxin exists in living centipedes, and the content is very small. Medicinal centipedes are processed dried whole insects, which basically do not contain centipede toxins. However, it is recorded in the literature that centipedes are slightly poisonous. Compared with living centipedes, the hemolytic activity of dried centipedes was greatly reduced, and the activity decreased with the prolongation of storage time.

In addition, the researchers also found that there is a big difference between the two in the promotion of platelet aggregation. The centipede's role in dredging collaterals and promoting blood circulation may be closely related to proteases, hemolytic factors and other blood system activating factors in its venom.

[0005] In the prior art, the methods for extracting the medicinal components of centipede toxin include enzymatic hydrolysis, water extraction and alcohol precipitation, alcohol extraction and alcohol precipitation, water leaching, etc., but the effective rate of the extracted active ingredients is not high.

Images