Kurooka S, Okamoto S, Hashimoto M
J Biochem. 1977 Feb;81(2):361-9. doi: 10.1093/oxfordjournals.jbchem.a131467.
A new and simple colorimetric method for human serum lipase [EC 3.1.1.3] assay has been developed, using 2,3-dimercaptopropan-1-ol tributyroate as a substrate, 5,5'-dithiobis(2-nitro-benzoic acid) as a chromogenic reagent, phenylmethylsulfonyl fluoride as an inhibitor of serum esterases, and sodium dodecylsulfate as a lipase activator. The method requires only 50 micron1X2 of serum sample and a reaction time of less than 30 min. The method is reproducible and sensitive enough to measure low levels of lipase activity in normal and abnormal sera. The gel filtration of serum samples on a Sephadex G-200 column gave one peak of lipase activity, when measured by the present method, and the molecular weight of the enzyme was identical with that of lipase of human pancreatic origin, confirming the specificity of this new method for the serum lipase.
已开发出一种用于测定人血清脂肪酶[EC 3.1.1.3]的新型简便比色法，该方法使用2,3-二巯基丙醇三丁酸酯作为底物，5,5'-二硫代双(2-硝基苯甲酸)作为显色剂，苯甲基磺酰氟作为血清酯酶抑制剂，十二烷基硫酸钠作为脂肪酶激活剂。该方法仅需50微升血清样本，反应时间少于30分钟。该方法具有可重复性，灵敏度足以测量正常和异常血清中低水平的脂肪酶活性。当用本方法测量时，血清样本在Sephadex G-200柱上进行凝胶过滤得到一个脂肪酶活性峰，且该酶的分子量与人胰腺来源的脂肪酶相同，证实了这种新方法对血清脂肪酶的特异性。
