Koncki Robert, Rudnicka Katarzyna, Tymecki Łukasz
University of Warsaw, Department of Chemistry, Pasteura 1, PL-02-093 Warsaw, Poland.
Anal Chim Acta. 2006 Sep 1;577(1):134-9. doi: 10.1016/j.aca.2006.05.100. Epub 2006 Jun 17.
A simple flow injection system for potentiometric detection of alkaline phosphatase (ALP) activity has been developed and adapted for determination of selected inhibitors of this enzyme. In this system monofluorophosphate (MFP) has been applied as a specific ALP substrate. The use of this substrate enables application of fluoride ion selective electrode (FISE) as a detector of the product of the enzyme catalyzed reaction. Moreover, chemical stability and low cost of MFP enables the use of the substrate as a component of the carrier. This way, fluoride ions contained in this substrate define and stabilize baseline signal generated by the detector. Effects from several potential ALP inhibitors and interfering species were studied and discussed. The system allows inhibitive detection of beryllium and vanadate ions at ppb levels with relatively high selectivity, short time of analysis and high throughput of the system (near 8 samples h(-1)).
已开发出一种用于电位检测碱性磷酸酶(ALP)活性的简单流动注射系统,并将其用于测定该酶的特定抑制剂。在该系统中,单氟磷酸酯(MFP)被用作特异性ALP底物。使用这种底物能够将氟离子选择性电极(FISE)用作酶催化反应产物的检测器。此外,MFP的化学稳定性和低成本使得该底物能够用作载体的组分。通过这种方式,该底物中所含的氟离子定义并稳定了检测器产生的基线信号。研究并讨论了几种潜在的ALP抑制剂和干扰物质的影响。该系统能够以相对较高的选择性、较短的分析时间和较高的系统通量(接近每小时8个样品)对ppb水平的铍离子和钒酸根离子进行抑制性检测。