羊软骨Ⅱ型胶原蛋白提取中脱钙、脱脂工艺的改进

LMPROVED PROCESS FOR DEGREASING AND DECALCIFICATION OF COLLAGEN TYPE Ⅱ FROM SHEEP CARTILAGE

以羊软骨为试验材料,对前处理工艺-脱脂、脱钙工艺进行了改进。经过单因素实验筛选确定了较为理想的脱脂剂为氯仿:甲醇:水=1:2:0.8,脱脂条件是温度为25℃、时间为48 h、固液比1:10时其脱脂率为最高,达55.88%;选择0.5mol/LEDTA为脱钙试剂时脱钙率最高,达29.16%;SDS-PAGE电泳结果表明:用前处理改进后提取的的胶原蛋白与sigma公司的Ⅱ型标准品相一致,证明羊软骨中存在的蛋白主要是Ⅱ型胶原蛋白。紫外光谱分析结果显示:所提胶原的最大吸收峰为230nm左右,与Ⅱ型标准品最大相一致。

The sheep cartilages were collected as raw materials, Optimization of degreasation and decalcification process during ex- tracting collagen from sheep cartilages were investigated. Improved process of degreasing and decalcification were confinued . The opti- mum parameters of degreasing were ： temperature 25~C , degreasing time 48h, chloroform ： methanol ： water = 1 ： 2：0.8. When ratio of Solvent to Solid was 1 ： 10 ,the ratio of degreasing was 55.88%. The optimum decalcifying agent was 0.5mol/L EDTA and the ratio of decalcification was 29.16 %. The extracted collagen with improved process was same with the protein and Sigma C - II by SDS - PAGE. The absorption peak of C - II in UV was about 230 nm. The results showed that the collagen type II was mainly protein in sheep cartilage.