Extraction, Isolation, Purification And Function Identifying Of Fish Head Polysaccharide

During the process of aquatic products,a lot of offals which occupy a large proportion of the fish body should be reused.The proportion could be as large as 40%-50%.Kinds of effort are taken to make a great use of those offals for some new products,which are necessary in the following fields:agriculture,light industry, environmental industry and food industry.As a result,people have paid more and more attention in the process of fish and looking forward to higher economy,zoology and social benefits.This study takes freshwater fish head as a raw material and do some research on its polysaccharide.The research includes:different extracting methods and various extracting conditions of fish head polysaccharide,in vitro anti-oxidation experiments and some basic cell culture work,through which more functions of fish head polysaccharide could be recognized further.Through some single factor optimization and uniform design experiments,five different extracting methods has been used to study fish head polysaccharide,SDS-PAGE has also been used to make sure which kind is the best one.NaOH extracting method is proved to be a better one than the four other methods and is also optimized to figure out the best extracting conditions:extracting temperature is 30℃,extracting time is 13 hours,the material fluid ratio is 1:55 and ethanol concentration is 75%.The results of in vitro anti-oxidation experiments prove:fish head polysaccharide has certain effects to eliminate DPPH and another two kinds of free radicals.Especially to DPPH,a higher cleaning away rate could be reached.The IC₅₀ value on O₂^-·and IC₅₀ value on·OH are separately 0.41mg/mL and1.98 mg/mL.The results of MTT indicate that fish head polysaccharide could restrain and control the multiplication of K562 and BEL7402.An evidence of the typical apoptotic morphologic changes in K562.On the contrary,fish head polysaccharide do have some effects to control the multiplication of BEL7402,but apoptosis is not the main approach.After fish head polysaccharide is acted on K562,as time goes by,cells in S phase obviously increases,the proportion of cells in G₀/G₁ and G₂/M phase decreases. BEL7402 acted by fish head polysaccharide does not appear a typical change of cell cycle.The proportions of apoptosis cell of K652 and bel7402 are 21.28%and 11.80%.