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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ＳｈｉｍＰａｃｋＣＬＣＯＤＳ柱（５μｍ），以甲醇—乙腈—００５％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
In order to ensure the correctness of caffeine metabolism probe, a method for the determination of urinary caffeine metabolites by RP-HPLC was established. ShimaPackCLC ODS column (5μm) was used in Shimadzu Japan. The mobile phase was methanol-acetonitrile-0.05% acetic acid (12:1:87). Caffeine and its 13 metabolites and internal standard could be separated well. Five of the major metabolites, AFMU, 1U, 1X, 17U and 17X, were investigated. The results showed that the recoveries of these five metabolites were above 87%, with the error of less than 3% between day and day, indicating the stability of the method. The urinary concentrations of five major metabolites in 120 volunteers were determined and the conditions were established to further evaluate the activity of various drug-metabolizing enzymes