高效液相色谱柱后反应安培法检测胞壁酸

Amperometric detection of muramic acid in high-performance liquid chromatography with a post-column reaction.

作者信息

Watanabe N

出版信息

J Chromatogr. 1984 Dec 21;316:495-500. doi: 10.1016/s0021-9673(00)96177-6.

Abstract

Highly sensitive detection of muramic acid has been developed by using an amperiometric detector in high-performance liquid chromatography. The reducing ability of muramic acid was coupled with the redox reaction of bis(1,10-phenanthroline)copper(II), CBP, as mediator. CBP, reduced by muramic acid in a post-column reaction, was reoxidized with the amperometric detector, resulting in a highly sensitive and selective detection. A doublet peak was obtained, due to the alpha- and beta-anomers of muramic acid. This was merged into a single peak by adding phosphoric acid to the sample solution. Addition of phosphoric acid to serum, spiked with muramic acid, also clarified the chromatogram. The detection limit was 0.05 ppm (4 pmol of muramic acid injected) in serum samples.

摘要

通过在高效液相色谱中使用安培检测器，已开发出对胞壁酸的高灵敏度检测方法。胞壁酸的还原能力与作为媒介物的双（1,10-菲咯啉）铜（II）（CBP）的氧化还原反应相耦合。在柱后反应中被胞壁酸还原的CBP，通过安培检测器重新氧化，从而实现高灵敏度和选择性检测。由于胞壁酸的α-和β-异头物，得到了一个双峰。通过向样品溶液中加入磷酸，该双峰合并为一个单峰。向添加了胞壁酸的血清中加入磷酸，也使色谱图更加清晰。血清样品中的检测限为0.05 ppm（注入4 pmol胞壁酸）。