Studies Of Synthetic Extraction Of Collagen And Hydroxyapatite From The Fish Scales Of Black Carp

In this study,the procedure for hot water extraction of collagen and making small molecular polypeptide by enzymatic hydrolysis from produced collagen, meanwhile,making hydroxyapatite from the enzymatic hydrolysised fish scale was developed.The fish scale was from fresh water fish species-the Black carp fish as the pilot case study.The whole research was aiming at developing a feasible prodedure for effective ultilization of fish scale.The main research results were as follows:1 The procedure for hot water extraction of collagen from the Fish Scales of Black CarpIt was researched by hot water extracting for extraction of collagen from the Fish Scales of Black Carp.Primary and secondary factors that it was affected extraction of collagen from the fish scales of black carp are as follows:the ratio of substrate and solvent＞extracting time＞extracting temperature.The optimal conditions of the extraction of Black carp fish collagen were found by orthogonal tests as follows: when the ratio of substrate and solvent is 1:14,the extracting time is 2.5h and the extracting temperature is 75℃.A spectrophotometric method for the content of the collagen in the Black carp fish scales was established and a regression equation was obtained as A =0.0629ρ+0.0064,the regression coefficient R²=0.9992.The experiments that was mensurated by mensurating the concentration of the hydroxide-proline through the colorimetry showed:the extracting rate is 70.71%,the determination of recovery rate is 97.33%.2 The preparation of the collagen from the Black carp fish scales by Enzymatic HydrolysisSingle factor and orthogonal experiments were used to analyze the main material and environmental factors which affect the preparation of collage by Enzymatic hydrolysis.The results showed that the optimal condition for the extraction technique,which were as follows:substrate concentration of 50%,temperature 54℃,alkaline proteinase 1.5%,pH value 10,hydrolysis time 4.5h,and the maximum hydrolysis degree was achieved as 28.81%.Through SDS-urea plastic electrophoresis,electrophoresis strip was quite smart, the molecular weight of the enzymatic collagen were isolated to 6KDa,8KDa,14KDa,20KDa,25KDa,30KDa.Then make use of(NH₄)₂SO₄ salting-out to concentrate,and remove sale with Sephadex G-25,during concentrating by rotary evaporator,finally freeze drying to prepare milky white collagens polypeptide production.3 The preparation of Hydroxyapatite from the Fish Scales of Black CarpHydroxyapatite(HAP) were extracted from fish scales by high-temperature sintering.Influence of sintering temperatures,sintering time and pH values of pre-treated solution on phase composition,structure and morphology of extracted particles was studied by X-ray diffraction.Results show that the pure HAP can be obtained after pre-treating waste of fish scales that it was remainde by extrating collagen in pH=13 pre-treated solution and calcining at 800℃for 4h,while the admixture ofβ-tricalcium phosphate(β-TCP) and HAP were obtained by sintering fish scales directly.