Study On Grass Carp Fish Scale Collagen Extraction And Purification

Collagen, rich in nutrients, contains18amino acids including seven kinds ofessential amino acids. Collagen is not only of high nutritional value, but also of highvalue, and that can be applied in the area of biomedical, food processing, health careproducts, etc. There are large amounts of collagen in fish scales and the productionof fish scale is up to more than three hundred thousand tons. To improve thecomprehensive utilization of fish scales of protecting of the environment andconserve resources is of great significance. This research extracts collagen withultra-high pressure auxiliary hot water and purifies the extracted collagen byultra-filtration. At last, Analysis and identification of the collagen are made bySDS-PAGE electrophoresis. The main research contents and findings are as follows:⑴The determination of extraction collagen with ultra-high pressure auxiliaryhot water: First, extract collagen with hot water after the fish scale treated byultra-high pressure, and do the5×5crossover experiment of two factors (theultra-high pressure time and the ultra-high pressure) with ANOVA and Duncan’s newmultiple range method, concluding the optimal ultra-high pressure time15minuetsand the optimal ultra-high pressure250MPa. Second, extract collagen with hot waterfrom fish scale treated by ultra-pressure, and investigate the effects of four factorswhich are hot water temperature, hot water extraction time, pH and solid-liquid ratioon extraction rate. Do the single factor experiment followed with orthogonal testnamely, concluding that the best hot water extraction process was extracted at90℃,2.5hours, pH=7.0, and solid-liquid ratio1:100. Under such circumstances, theextraction ratio was87.9%. Third, compares the two processes, extraction collagenwith ultra-high pressure auxiliary hot water and traditional extraction with water, theformer can greatly improve the extraction rate of collagen with simple method andshort time.⑵SDS-PAGE Analysis: analyze the crude collagen liquid with SDS-PAGEfrom the extraction process of ultra-high pressure auxiliary hot water under the bestconditions, resulting that the collagen was of three significant characteristics chains but not pure enough. Purification was needed.⑶The determination of ultrafiltration separation to purify the crude collagenliquid: First, the research uses the ultra-filtration membrane which aperture size was100kDa to separate and purify the crude collagen water. Second, based on the singlefactor study, to investigate the condition parameters of ultrafiltration separationprocess with quadratic rotation-orthogonal combination tests, and get the regressionmodel of retention rate(Y) and operating pressure(A),operating temperature(B) andpH(C). The regression equation is as follows:Y95.61-6.86A1.43B0.72C0.59AB4.12A~24.11B~20.69C~2Third, analyzes the research ultra-filtration process parameters on the retentionrate, concluding the optimization condition is operating pressure0.14MPa, operatingtemperature40℃, pH=9.50.⑷SDS-PAGE Analysis: analyze the concentrated solution of purified collagenwith SDS-PAGE. The extracted collagen was identified to be protein collagen withthe electrophoretic pattern, which is of obvious features of three clear peptide chainswithout other band.