Method for refining bee venom

Abstract

A method for refining bee venom comprises the steps: preparing the bee venom, adding water of 5-10 times of the amount of the bee venom into the bee venom to perform dissolution and then perform high speed centrifugation, and water phase is obtained after the centrifugation; adding acetone with the 1-3 times of the volume of the water phase into the water phase to perform standing at the temperature of less than or equal to -8 DEG C until the precipitation finishes completely, adding the acetone in precipitate and smashing the mixture after separation, performing filtering separation and volatilizing the acetone in the precipitate to obtain a rough bee venom product; using 0.08-0.12mol / L of acetum to dissolve the rough product, then using a sephadex G50 chromatographic column to perform the separation and purification, using acetum with the same concentration to perform elution and using an ultraviolet detector to perform on-line detection at 280nm of wavelength, collecting eluent of a third peak, adding activated carbon to perform slow stirring, and freezing and drying filtrate after the separation. In the refined bee venom, melittin content is greater than or equal to 76%, macromolecule substance content is less than 1.5%, and a lyophilized agent for clinical injection can be directly added.

Description

1. Technical field [0001] The invention relates to a method for separating and purifying bee venom, in particular to a method for refining bee venom. 2. Background technology [0002] Bee venom is a kind of venom with aromatic smell secreted by worker bee glands. It is stored in the poison sac and is discharged from the tail sting needle when worker bees attack other species. It is a complex mixture with complex chemical composition. Peptides, large molecular weight enzymes, small molecular weight amines and other substances, peptides mainly include melittin (accounting for about 50% of the dry weight of bee venom), melittin, MCD peptide, heart peptide, andu peptide, Histamine peptides, etc. Enzymes include more than 55 kinds of enzymes, of which phospholipase A 2 (referred to as PLA 2 , average molecular weight 14500) is the main substance that produces allergic reactions after being stung by bees. Amines include other active amines such as histamine and catecholamines. A...

AI Technical Summary

Problems solved by technology

[0003] Bee venom has a long history of being used in folk health care. Modern research has confirmed that bee venom has anti-inflammatory and analgesic effects, and is used to treat rheumatoid arthritis, rheumatoid arthritis, ankylosing spondylitis and other rheumatic diseases. Inflammatory muscle neuralgia and other diseases have a definite curative effect, however, the large molecular weight enzymes in bee venom are likely to cause allergies, and the small molecular weight amines are easy to cause pain, which greatly affects the clinical promotion and use of bee venom preparations

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