目的：证明响应曲面法（ RSM）是确认淀粉酶（ AMY）参考测量程序最佳测量条件的方法。方法采用单因素方差分析研究pH值、氯离子（ Cl－）、亚乙基-P-硝基苯-D-麦芽七糖苷（ EPS）浓度对AMY活性的影响是否具有统计学意义。采用Box-Behnken试验设计，在实验室条件下研究pH值（6．9～7．1）、Cl－（50～90 mmol／L）和EPS浓度（4．5～5．5 mmol／L）对AMY活性的联合效应，建立pH值、Cl－和EPS对AMY活性影响的定量关系模型，优化得出pH值、Cl－和EPS的最佳组合。结果 pH值、Cl－和EPS对AMY活性影响有统计学意义（ P＜0．05）；pH值、Cl－和EPS对AMY活性影响的一次、二次效应均有统计学意义（ P＜0．01），Cl－和EPS交互作用有统计学意义（P＜0．05）。 AMY最优条件：pH值7．02、Cl－72．79 mmol／L、EPS 5．07 mmol／L。结论所得pH值、Cl－和EPS的最佳测量条件与国际临床化学和检验医学联合会（ IFCC）推荐条件无差异，RSM可以作为确认参考测量程序最佳测量条件的方法。%Objective To confirm that response surface method ( RSM) is a method for amylase ( AMY) reference determination procedure with optimum conditions.Methods The single factor analysis of variance was used to study pH, chloride ion (Cl-) and ethylidene-4-nitrophenyl-alpha-D-maltoheptaoside(EPS) concentrations in order to know whether they had statistical significance for AMY activity.Box-Behnken design was used to study the combined influence of pH (6.9-7.1), Cl-(50-90 mmol/L) and EPS (4.5-5.5 mmol/L) concentrations on AMY activity.The quantitative relation model on pH, Cl-and EPS concentrations for AMY activity was established in order to obtain their optimum combination.Results There was statistical significance on the influence of the 3 factors on AMY activity ( P<0.05) . The one-shot and second-order effects of pH,Cl-and EPS had statistical significance on AMY activity (P<0.01).Cl-and EPS interacted significantly (P<0.05).The optimum conditions for AMY were pH 7.02,Cl-72.79 mmol/L and EPS 5.07 mmol/L.Conclusions The optimum conditions of pH, Cl- and EPS have no difference with those recommended from the International Federation of Clinical Chemistry and Laboratory Medicine ( IFCC ) .RSM is a method with optimum conditions to confirm the reference determination procedure.
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