HPLC法间接测定肿瘤患者外周血中二氢尿嘧啶脱氢酶的活性  

Indirect Determination of Dihydropyrimidine Dehydrogenase Activity in Peripheral Blood of Cancer Patients by HPLC

作　　者：姚勤[1] 邓杨林[1] 吴迪[1] 陈镜宇[2] 张峻[1] 

机构地区：[1]昆明医学院第一附属医院临床药学中心,昆明650032 [2]昆明医学院,昆明650500

出　　处：《中国药房》2012年第22期2045-2046,共2页China Pharmacy

基　　金：云南省应用基础研究面上项目(2009CD085)

摘　　要：目的:建立间接测定肿瘤患者外周血中二氢尿嘧啶脱氢酶(DPD)活性的方法。方法:采用高效液相色谱(HPLC)法测定肿瘤患者血浆中内源性物质二氢尿嘧啶(H2U)和尿嘧啶(U)的比值,间接估算DPD的活性。其中,色谱柱为XterraRPC18,流动相为甲醇-0.02mol·L-1磷酸二氢钠(pH4.0)(5:95),流速为1.0mL·min-1,检测波长分别为204nm(H2U)、260nm(U)。结果:H2U、U检测浓度分别在62.5～2000μg·L-1(r=0.9997)、15.625～500μg·L-1(r=0.9993)范围内线性关系良好,日内RSD分别≤7.7%、8.3%,日间RSD分别≤13.8%、13.4%,方法回收率分别为99.63%～103.02%、98.38%～109.08%。63例肿瘤患者的DPD活性(H2U/U比值)在肿瘤患者中变异较大,范围为1.51～6.67。结论:本方法可用于肿瘤患者外周血DPD活性的常规监测。OBJECTIVE：To establish a method for the determination of dihydropyrimidine dehydrogenase （DPD） activity in peripheral blood of cancer patients indirectly.METHODS：The ratio of dihydrouracil （H2U） to uracil （U） in plasma of cancer patients was determined by HPLC,and the activity of DPD was predicted indirectly.The determination was performed on Xterra RPC18 column with mobile phase composed of methanol-0.02 mol·L-1 sodium dihydrogen phosphate （pH 4.0）（5：95） at the flow rate of 1.0 mL·min-1.The detection wavelength was 204 nm for H2U and 260 nm for U.RESULTS：The linear ranges were 62.5～2 000 μg·L-1（r=0.999 7） for H2U and 15.625～500 μg·L-1 for U（r=0.999 3）.The RSDs of intra-day and inter-day were less than 7.7% and 13.8% for H2U 8.3% and 13.4% for U.The recoveries of H2U were 99.63%～103.02% and those of U were 98.38%～109.08%.The activity of DPD in 63 cancer patients （H2U/U ratio） varied greatly in the range of 1.51～6.67.CONCLUSION：The method can be used for routine monitoring of DPD activity in peripheral blood of cancer patients.

关 键 词：二氢嘧啶脱氢酶 尿嘧啶 二氢尿嘧啶 高效液相色谱法 

分 类 号：R969.1[医药卫生—药理学] R979.1[医药卫生—药学]