气相色谱法测定脂肪酶的活力Determinationoflipase.pdf

628 药学学报 Acta Pharmaceutica Sinica 2009, 44 (6) : 62863 1 气相色谱法测定脂肪酶的活力 * 李谭瑶, 邓克国, 陈 波, 姚守拙 (湖南师范大学化学生物学及中药分析教育部重点实验室, 湖南 长沙 410081) 摘要: 本文发展了一种以气相色谱检测脂肪酶活力的方法。以三丁酸甘油酯为底物，通过气相色谱对产物正 丁酸进行定性定量分析。正丁酸在0.11～11.35 mmol L 1 内线性关系良好, 回归方程为: y = 45 464x 3 115.2, r = 0.996 8 。反应时间仅需5 min ，最适pH 值和温度分别为7.5 和32 。低、中、高3 个 度正丁酸的回收率分别为 90.3%，104.6%和89.4% ；RSD 分别为3.01%，4.50%和6.64% 。通过实验测定出Km = 0.25 mmol mL 1，奥利司他 的半数抑制 度 (IC50) 为0.048 5 mg mL 1。本方法具有反应体积小、操作简单、快速、灵敏等特点。 关键词: 脂肪酶；三丁酸甘油酯；正丁酸；气相色谱 中图分类号: R917 文献标识码:A 文章编号: 05 13-4870 (2009) 06-0628-04 Determination of lipase activity by gas chromatography * LI Tan-yao, DENG Ke-guo, CHEN Bo , YAO Shou-zhuo (Key Laboratory of Chemical Biology and Traditional Chinese M edicine Research, M inistry of Education, Hunan Normal University, Changsha 410081, China) Abstract: A rapid gas chromatography method was developed for determination of lipase activity using tributyrin as substrate. The standard curves of butyric acid hydrolyzed from tributyrin were linear in the range of 0.11 11.35 mmol L 1. The recoveries of low, moderate and high concentrations of tributyrin were 90.3%, 104.6%, 89.4% with RSD of 3.0 1%, 4.50%, 6.64%, respectively. The incubation time was only 5 minutes which was less than with the half time of the conventional titrimetry and spectrophotometry. The optimum pH value was 7.5 and the optimum temperature was 32 ℃. B