COMPOSITIONS FOR LIPASE ACTIVITY DETERMINATION AND METHOD OF DETERMINING ACTIVITY
申请人:Asahi Kasei Pharma Corporation
公开号:EP1813680A1
公开(公告)日:2007-08-01
[PROBLEMS] To provide lipase activity determination reagents which function by an enzymatic method. The reagents are easy to use in an ordinary clinical examination, have excellent handleability, and are excellent in accuracy and reproducibility.
[MEANS FOR SOLVING PROBLEMS] Lipase activity is determined with any of reagents which comprise a low-concentration buffer and a diglyceride dissolved therein. The diglyceride is used as a substrate for lipase, whereby the liquid reagents can have long-term storage stability. One of the reagents converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, pyruvate kinase, lactate dehydrogenase, reduced NAD, ATP, and phosphoenol pyruvate. Another reagent converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, glucose, ADP-dependent hexokinase, glucose-6-phosphate dehydrogenase, oxidized NAD or oxidized NADP, and ATP. A further reagent converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, glycerol-3-phosphate oxidase, peroxidase, and a dye which colors in the presence of hydrogenperoxide.
[问题] 提供用酶法测定
脂肪酶活性的试剂。这些试剂易于在普通临床检查中使用,具有极好的可操作性,而且准确性和再现性极佳。
[解决问题的方法]
脂肪酶活性的测定可使用由低浓度缓冲液和溶于其中的二
甘油酯组成的试剂。二
甘油酯用作
脂肪酶的底物,因此液体试剂具有长期储存稳定性。其中一种试剂借助单
甘油酯
脂肪酶将
脂肪酶反应产生的单
甘油酯转化为
甘油,并进一步含有
甘油激酶、
丙酮酸激酶、
乳酸脱氢酶、还原型
NAD、
ATP 和
磷酸烯醇
丙酮酸。另一种试剂在单
甘油酯
脂肪酶的帮助下将
脂肪酶反应产生的单
甘油酯转化为
甘油,并进一步含有
甘油激酶、
葡萄糖、
ADP 依赖性
己糖激酶、
葡萄糖-6-
磷酸脱氢酶、氧化的
NAD 或氧化的
NADP 和
ATP。另一种试剂在单
甘油酯
脂肪酶的帮助下,将
脂肪酶反应产生的单
甘油酯转化为
甘油,并进一步含有
甘油激酶、
甘油-3-
磷酸氧化酶、
过氧化物酶和一种在
过氧化氢存在下着色的
染料。