Principles of stopped-flow sequential injection analysis and its application to the kinetic determination of traces of a proteolytic enzyme.
Principles of stopped-flow sequential injection analysis and its application to the kinetic determination of traces of a proteolytic enzyme.
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停流连续注射分析原理及其在蛋白水解酶痕量动力学测定中的应用。
DOI：
10.1021/ac00017a006
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发表时间：
1991
影响因子：
7.4
通讯作者：
Gübeli,T
中科院分区：
文献类型：
--
作者：
Ruzicka,J;Gübeli,T
描述了顺序注射分析的概念，并演示了这种新技术在蛋白水解酶痕量测定中的应用。自动化方法基于带有荧光检测的停流程序。采用了由凸轮驱动柱塞泵产生的正弦流型，而不是传统的恒定流量。使用非反应性荧光化合物的新型流动注射系统的验证在3个数量级的浓度范围内产生了高度可重复性的结果。在5 ~ 75 pu mL~ 1范围内，酶活的标定图可以用二阶多项式拟合。检测Rmlt为2.5/uu mL~\，对应于7.2 ng mL~ 1纯酶。
The concept of sequential Injection analysis Is described, and an application of this novel technique to an assay of traces of a proteolytic enzyme Is demonstrated. The automated method Is based on a stopped-flow procedure with fluores-cence detection. Instead of using the conventional constant flow rate, a sinusoidal flow pattern, as generated by a robust cam-driven piston pump, has been applied. The validation of the novel flow Injection system using a nonreactlve fluorescent compound yielded highly reproducible results over a concentration range of 3 orders of magnitude. The calibration graph for the enzyme activities In the range 5-75 pu mL~ 1 could be fitted a second-order polynomial. The detection Rmlt Is 2.5/uu mL~\which corresponds to 7.2 ng mL~ 1 pure active enzyme.