Optimization Of The Extraction Process Of Rat Tail Tendon Collagen By Orthogonal Experiment Method
Keywords
Rat Tail Tendon, Collagen, Collagen Sponge, Extraction Process, Orthogonal Test Method
Abstract
Objective: To explore the effects of extraction time, acetic acid concentration and solid-liquid ratio on the extraction of rat tail tendon collagen, establish and optimize the extraction process of rat tail tendon collagen, and improve the collagen extraction efficiency. Methods: Healthy SD rat tails were selected, and the acid dissolution method was used to extract rat tail tendon collagen. Different extraction times (12, 24, 48, 72 and 96 hours) and acetic acid concentrations (0.10, 0.25, 0.50, 0.75 and 1.00 mol) were observed. ¡¤L-1) and the effect of solid-liquid ratio (g:mL) (1:5, 1:10, 1:20, 1:40 and 1:80) on the collagen extraction rate, using orthogonal experiments to determine the extraction time , acetic acid concentration, material-liquid ratio and other influencing factors. The extracted collagen was identified through SDS-PAGE electrophoresis experiment, tannic acid precipitation experiment and Fourier transform infrared spectroscopy experiment. Results: Single factor analysis showed that the collagen extraction rate increased with the increase of extraction time, acetic acid concentration and material-liquid ratio within a certain parameter range. Through orthogonal experiments, the optimal process parameters for rat tail tendon collagen extraction were extraction time 72 h, acetic acid concentration 0.5 mol¡¤L-1, and solid-liquid ratio 1:40. SDS-PAGE electrophoresis showed that the sample was type I collagen, the tannic acid precipitation experiment showed that the degree of collagen degradation was low, and the Fourier transform infrared spectrum showed that the collagen structure was intact. The determination of the amino acid content of the sample is consistent with the composition characteristics of collagen. Neutral collagen solution can form a solid collagen gel at 37¡ãC. After the collagen gel is cross-linked with carbodiimide (EDC), scanning electron microscopy shows that the collagen gel is internally connected to form a honeycomb porous structure with moderate pore size. Conclusion: The extraction process of rat tail tendon collagen was successfully established and optimized, and the optimal process parameters of factors such as extraction time, acetic acid concentration, and material-liquid ratio were obtained, which effectively improved the extraction efficiency of rat tail tendon collagen.
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Original research was done by Song Weiwei, Liu Guomin, Yuan Haibo, Peng Chuangang, Xu Kun, Yin Dehui, Cao Hongqian, Li Juan
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