Meek J L, Eva C
J Chromatogr. 1984 Dec 28;317:343-7. doi: 10.1016/s0021-9673(01)91673-5.
Enzymes can be used in high-performance liquid chromatography post-column reactors to improve sensitivity and specificity of detection for some compounds by converting the compounds to easily detectable products. The enzymes can be covalently bound to a post-column reactor, but a simpler approach is to bind them by adsorption to an ion exchanger or a hydrophobic interaction support. This technique has been applied to electrochemical detection of acetylcholine by using adsorption of choline oxidase and cholinesterase to a 3-cm long commercially available weak anion-exchange cartridge. Conversion of acetylcholine to peroxide is quantitative during the 10-sec residence time in the cartridge. Enzyme elution from the cartridge is negligible when low ionic strength mobile phases are used. Fresh enzyme needs to be added to the cartridge at only 1-2 week intervals.
酶可用于高效液相色谱柱后反应器，通过将某些化合物转化为易于检测的产物来提高检测的灵敏度和特异性。酶可以共价结合到柱后反应器上，但一种更简单的方法是通过吸附将它们结合到离子交换剂或疏水相互作用载体上。该技术已应用于乙酰胆碱的电化学检测，即将胆碱氧化酶和胆碱酯酶吸附到一个3厘米长的市售弱阴离子交换柱上。在柱中10秒的停留时间内，乙酰胆碱向过氧化物的转化是定量的。当使用低离子强度流动相时，酶从柱上的洗脱可以忽略不计。只需每隔1-2周向柱中添加新鲜酶即可。
