咖啡因5种主要代谢物的反相高效液相色谱法测定

为保证应用咖啡因代谢探针的正确性,建立了反相高效液相色谱测定尿中咖啡因代谢物的方法。采用日本岛津 Shim Pack CLC-ODS柱(5μm),以甲醇—乙腈—0.05%醋酸(12∶1∶87)为流动相,咖啡因和它的13种代谢物及内标均能良好分离。重点对其中5个主要代谢物AFMU,1U,1X,17U,17X进行了测定。结果表明这5种代谢物回收率均在87%以上,日内和日间误差均小于3%,显示了方法的稳定性。并对120例志愿者尿中5种主要代谢物浓度进行了测定,为进一步评价多种药物代谢酶活性创造条件。

HPLC DETERMINATION OF FIVE CAFFEINE METABOLITES

An HPLC method for the determination of caffeine metabolites in urine was established. Shim Pack CLC-ODS column (5 microns) was eluted with the mobile phase of methanol--acetonitrile--0.05% acetic acid = 12:1:87 (v/v) at a flow rate of 1.2 ml.min-1, and the ultraviolet absorbance was monitored at 280 nm. The 13 caffeine metabolites and caffeine were well separated and the concentrations of the five metabolites, AFMU, 1U, 1X, 17U, and 17X, were determined. The recoveries of the five metabolites were above 87%, the inter- and intra-day variations were less than 3%. The concentrations of the five metabolites in 120 volunteers were determined. The ratios of the metabolites were employed for the assessment of CYP1A2, NAT, and XO enzymes successfully.