The Technology Research Of Leech Processed With Wine

ObjectiveTo explore the difference of protein and amino acid in leech and its products processed with wine,provide the scientific basis for the connotation of processing technology.MethodsAntithrombin activity of leech and its processed products was detected according to the Leech identification method and determination methods in "Chinese Pharmacopoeia" version 2015.The total content of free amino acids of leech and its processed products was determined by ninhydrin method.7 kinds of free amino acids of leech and its processed products was determined by high-performance liquid phase chromatography with pre column derivative method.The crude extraction of protein was collected by saturated ammonium sulfate precipitation,TCA/acetone precipitation and pyrolysis liquid.The sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)was applied to evaluate the three methods and select the best protein extraction method by protein species richness.And construct the Etablish of(2-DE)electrophoresis system of leech and its processed products.ResultsLeech and its product reached the standard of Pharmacopoeia requirements of identification and content,but the antith rombin activity of leech deceased after processed with wine.Ninhydrin method results show that in the total content of free amino acids of leech were obviously more than those of leech processed with wine.High performance liquid phase chromatography with pre column derivative method results show that the content of glutamic acid and phenylalanine were increased and the remaining five amino acids were decreased after the leech processed with wine.The results of SDS-PAGE indicated that the protein types of leech were more than those of leech processed with wine.The extraction with pyrolysis liquid can get more protein,which can be used as the protein extraction method of 2-DE.A protein differences spectrum has been obtained through 2-DE.PDQuest8.0 image analysis software show that 19 different protein spots were found,8 of which is down-regulated and 11 of which is up-regulated.The protein information through 2-DE can be used for protein markers specification processing technology,which can make sure the stability of the traditional process of leeches.ConclusionsAlthough wine processing technology make leech crunchy and smell drop is small,but the antithrombin activity,the type and quantity of protein and the total free amino acid content of leech processed with wine were reduced.If the protein is the main active ingredient leeches,wine processing technology is difficult to highlight its advantages.