Liquid chromatographic separation and on line bioluminescence…液相色谱分离和在线生物发光.pdfVIP

CLIN. CHEM. 26/6, 712-717 (1980) Liquid-Chromatographic Separation and On-Line Bioluminescence Detection of Creatine Kinase lsoenzymes William D. Bostick, Mark S. Denton, and Stanley R. Dinsmore We separated isoenzymes of creatine kinase by anion- zymes, with reagent addition and on-line detection of the exchange chromatography, with use of an elution gradient coupled NADPH reaction product, is an alternative approach containing lithium acetate (0.1 to 0.6 mol/L). A stream to electrophoresis. Nonreferenced photometric and fluoro- splitter was used to divert a 5% side stream of column metric detection of the column effluent (30, 31) offers no effluent, which was subsequently mixed with the reagents compensation for endogenous light-absorbing or fluorescing necessary for bioluminescence assay of the separated components in serum. Recently, we developed a continuously isoenzymes. The use of the stream splitter greatly de- referenced on-line detection system with self-blanking creased the rate of consumption of reagent and, when capabilities (32-34) for monitoring the NADPH reaction combined with a peristaltic pumping system, permitted product of the coupled assay. independent control of the side-stream flow rate. Thus In this paper, we investigate the utility of the firefly biolu-