Abstract:

Objective To investigate the effects of extraction time,concentration of acetic acid and solid-liquid ratio on the extraction of rat tail tendon collagen,and to establish and optimize the extraction process of rat tail tendon collagen,and to increase the extraction efficiency. Methods The healthy SD rats tail were selected,the rat tail tendon collagen was extracted by acid-soluble method,the influence of different extraction time (12,24,48,72,96 h),acetic acid concentrations (0.10,0.25,0.50,0.75,1 mol·L^-1) and solid-liquid ratios (1:5,1:10,1:20,1:40,1:80) in the extraction ratio of collagen was observed. The optimal extraction parameters were explored by orthogonal test to judge the extraction time,the concentration of acetic acid and solid-liquid ratio,etc. The extracted collagen was identified by SDS-PAGE electrophoresis,tannic acid precipitation test and Fourier transform infrared spectroscopy test etc. Results The univariate analysis results showed that the collagen extraction rates were increased with the increasing of the extraction time,the concentration of acetic acid and the liquid ratio within a certain range of parameters. The optimal parameters were the extraction time of 72 h,the concentration of acetic acid of 0.5 mol·L^-1,the solid-liquid ratio of 1:40.SDS-PAGE electrophoresis showed that the samples were mainly collagen typeⅠ.Tannic acid precipitation test showed a low degree of degradation of collagen.The extract was confirmed by infrared spectral analysis to be typeⅠcollagen with intact structure.After cross-linking of collagen,the SEM figure showed that the internal interconnection formed a porous honeycomb structure,and the pore size was suitable. Conclusion The extraction process of rat tail tendon collagen is successfully established and optimized.The optimum process parameters of the extraction time,the acetic acid concentration and the material liquid ratio are gained,which can improve the extraction efficiency of rat tail tendon collagen.

Key words: rat tail tendon, collagen, collagen sponge, extraction process, orthogonal test