单一稳定液体试剂直接测定α-淀粉酶

Direct Assay of α-Amylase by Single Stable Liquid Reagent

目的 :建立以α- 2 -氯 - 4-硝基苯 -半乳糖吡喃麦芽糖苷 ( Gal- G2 - CNP)作底物的α-淀粉酶 (α-amylase)的直接测定方法。方法 :利用国外最新合成的 Gal- G2 - CNP( alpha- 2 - chloro- 4- nitrophenyl-galactopyranosylmaltoside)作底物 ,用速率法测定α-淀粉酶活力。结果 :线性范围可达 30 0 0 U/L ( r=0 .9998) ,变异系数为 0 .30 %～ 0 .53%。与宝灵曼公司的 ET- G7- PNP法 〔1〕的相比 :r=0 .9998,Y=0 .76X+ 0 .2 0 ( n=2 0 0 ) ,与 G3 - CNP的相比 r=0 .9999,Y=1 .0 1 X+ 1 1 .7( n=2 0 0 )。结论 :简便、直接、灵敏、经济、试剂单一稳定、反应转化率高、线性范围宽、基质本身自然水解极微 ,各项指标符合国际临床化学学会( IFCC)提出的“理想的淀粉酶检测方法”的要求〔2〕。

Objective To establish a simple,sensitive,direct,wide line limit and single stable liquid reagent and kinetic rate assay method for alpha 2 chloro 4 nitrophenyl galactopyranosylmaltoside (Gal G 2 CNP) by the novel substrata.Method The novel substrata was prepared by the method of Gal G 2 CNP kinetic characters on overseas the latest substrata were studied.The kinetic rate assay method was evaluated.Results The linearity was arrived 3 000 U/L( r =0 999 8) and CV was 0 30%～0 53%.Compared with method of ET G 7 PNP(EPS) 〔1〕 for Boehringer Mannheim,it was shown that r =0 999 8, Y =0 76 X +0 20( n =200),with G 3 CNP, r =0 999 9, Y =1 01 X +11 7( n =200).Conclusion The direct assay method of amylase with a single stable liquid reagent was a simple,direct,sensitive and economic.It had high inversion of reaction,wide linearity range and the lowest of natural hydrolysis for the substrata itself.Its all index was accord with “the ideal method of assay for amylase”raised by IFCC 〔2〕 .It can performed with various types of automated analyzers.