用猪胰α-淀粉酶测定血清和尿液中钙的酶法。

Enzymatic method for assaying calcium in serum and urine with porcine pancreatic alpha-amylase.

作者信息

Kayamori Y, Katayama Y

机构信息

Department of Clinical Chemistry, National Cardiovascular Center, Osaka, Japan.

出版信息

Clin Chem. 1994 May;40(5):781-4.

PMID:8174252

Abstract

We developed a kinetic assay for calcium in serum and urine, based on the activation of porcine pancreatic alpha-amylase (EC 3.2.1.1) with 2-chloro-4-nitrophenyl-alpha-maltotrioside as substrate. The kinetic generation of 2-chloro-4-nitrophenol, monitored at 405 nm, was proportional to the concentration of calcium in serum and urine. The assay was developed and evaluated with the Cobas Bio centrifugal analyzer. The average within-run and between-day imprecision (CV) was 0.96/1.26% and 1.07/1.63%, respectively, for serum calcium and 1.50/2.54% and 1.70/2.64%, respectively, for urine calcium. Results of the proposed method (y) correlated well with those determined by atomic absorption spectrophotometry (x): y(serum) = 1.005x + 0.028 mmol/L (Sy/x = 0.058, r = 0.974, n = 50), and y(urine) = 1.017x-0.115 mmol/L Sy/x = 0.30, r = 0.981, n = 25). We also present data showing that the method is highly sensitive, rapid, relatively free of interference, and amenable to automation.

摘要

我们基于以2-氯-4-硝基苯基-α-麦芽三糖苷为底物激活猪胰腺α-淀粉酶（EC 3.2.1.1），开发了一种用于血清和尿液中钙的动力学测定方法。在405nm处监测到的2-氯-4-硝基苯酚的动力学生成与血清和尿液中钙的浓度成正比。该测定方法是使用Cobas Bio离心分析仪开发并评估的。血清钙的批内和批间平均不精密度（CV）分别为0.96/1.26%和1.07/1.63%，尿钙的批内和批间平均不精密度（CV）分别为1.50/2.54%和1.70/2.64%。所提出方法的结果（y）与原子吸收分光光度法测定的结果（x）相关性良好：y（血清）=1.005x + 0.028 mmol/L（Sy/x = 0.058，r = 0.974，n = 50），y（尿液）=1.017x - 0.115 mmol/L（Sy/x = 0.30，r = 0.981，n = 25）。我们还提供数据表明该方法具有高灵敏度、快速、相对无干扰且适合自动化。