摘要
建立了用亚乙基封闭的Ｇ７－ｐＮＰ（ＥＰＳ）作底物并偶联多功能α－葡萄糖苷酶测定α－淀粉酶（Ａｍｙ）的方法学。ＥＰＳ的稳定性好，工作试剂贮２～８℃可稳定２１天；室温（３０℃）稳定７天。由于多功能的α－葡萄糖苷酶对所有的ＥＰＳ残基（Ｇ１～Ｇ５）有相同的水解力，使色团ｐＮＰ１００％回收，通过计算ｐＮＰ的摩尔消光系数容易做到标准化。本法的延滞相较短（＜９０秒），零级反应时间＞１０分钟，线性范围达２０００Ｕ／Ｌ，分析总ＣＶ＜３．０％，本法（Ｙ）与ＢＭ公司的试剂（Ｘ）比较：ｒ＝０．９９８８，Ｙ＝１．０５４Ｘ－６．６。
Studied a methodolog for measuring α Amylase activity with Ethylidene Blocked G7 pNP(EPS) as substrate and with multifunctional α Glucosidase as ancillary enzyme. The stability of EPS was excellent, and the working reagent was stable for 21 days at 2 to 8℃. 100% recovery of the chromophore was due to the multifunctional α Glucosidase cleaves all EPS residues (pNPG1￣pNPG5) with similar velocity. Full cleavage of the substrate also means that standardization can easily be done by calculation with the molar absorbance of NPN. The lag phase of this test was <90 seconds; linear kinetic was >10 minutes; the linear range up to 2000 U/L. The total impresision (CV) was <3 0%. Comparison with BM's reagent (X), yieled a regression equation of Y=1 054X-6 6,r=0 9988
出处
《临床检验杂志》
CAS
CSCD
北大核心
1997年第1期5-8,共4页
Chinese Journal of Clinical Laboratory Science
