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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0.11～11.35mmol·L-1内线性关系良好,回归方程为:y=45464x-3115.2,r=0.9968。反应时间仅需5min,最适pH值和温度分别为7.5和32℃。低、中、高3个浓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=0.25mmol·mL-1,奥利司他的半数抑制浓度(IC50)为0.0485mg·mL-1。本方法具有反应体积小、操作简单、快速、灵敏等特点。
This paper developed a method for the detection of lipase activity by gas chromatography. Using butyric acid as substrate, the product n-butyric acid was qualitatively and quantitatively analyzed by gas chromatography. N-butyric acid in 0.11 ~ 11.35mmol·L-1 within a good linear relationship, regression equation: y = 45464x-3115.2, r = 0.9968. The reaction time only 5min, the optimum pH and temperature were 7.5 and 32 ℃. The recoveries of n-butyric acid at low, medium and high concentrations were 90.3%, 104.6% and 89.4%, respectively. The RSDs were 3.01%, 4.50% and 6.64%, respectively. Km = 0.25mmol · mL-1 was determined experimentally, and the half-value inhibition concentration (IC50) of orlistat was 0.0485mg · mL-1. The method has the advantages of small reaction volume, simple operation, quickness and sensitivity.