HPLC同时直接进样测定尿中咖啡因5种代谢物  

Determination of five metabolites of caffeine in urine by HPLC with direct sample injection method

作　　者：李军[1] 彭向前[2] 张鉴[1] 

机构地区：[1]山东省立医院临床药理中心,济南250021 [2]山东大学药学院,济南250012

出　　处：《药物分析杂志》2006年第4期480-482,共3页Chinese Journal of Pharmaceutical Analysis

基　　金：山东省自然科学基金(项目编号:Q99C05)

摘　　要：目的: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。方法:采用Shim-pack VP-COD 柱(4.6 mm×150 mm,5 μm),岛津 Shim-pack C_(18)预柱,流动相为乙腈-0.05%醋酸,0～24 min 时0.05%醋酸的量从2.5%线性增加至8%,流速为1 mL·min^(-1),柱温25℃,检测波长为280 nm。结果:咖啡因5种代谢物均能良好分离,在所考察的范围内有良好的线性,r 为0.9998～0.9999,平均回收率95.59%～102.2%,日内和日间误差均小于3%。结论:本方法简便、准确、快速,适合于尿中咖啡因代谢物的测定及 N-乙酰基转移酶、细胞色素 P450酶1A2和黄嘌呤氧化酶等药物代谢酶活性的研究。Objective：To establish an HPLC with direct injection method of metabolites of caffeine in the urine for assuring accuracy of caffeine used as metabolic probe. Method ：The Shim -pack VP -COD column（4. 6 mm × 150 mm ,5μm） and Shim - pack C18 pre - column was used. The mobile phase was acetonitrile -0. 05% acetic acid and the gradient program was 0. 05% acetic acid 2. 5% to 8% in 0 to 24 min. The flow rate was 1 mL · mim^ -1 ,and the column temperature was 25 ~C. The detective wavelength was at 280 nm. Results ：5 kinds of metabolites of caffeine could be separated completely. There was a good linearity （ r = 0. 9998 - 0. 9999 ）. The average recovery rate was in the range of 95.59% -102. 2%, the intra- day RSD and inter- day RSD were less than 3%. Conclusion：The method is simple, accurate and rapid, suitable for the determination of metabolites of caffeine in urine. The method is used to assay the activities of polymorphic N- acetyhransferase, cytochrome P450 1 A2 and xanthine oxidase.

关 键 词：反相高效液相色谱法 尿液 咖啡因代谢物 

分 类 号：R917[医药卫生—药物分析学]