HPLC法间接测定肿瘤患者外周血DPD的活性  

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

作　　者：周石桥 

机构地区：[1]曲靖市第五人民医院,云南曲靖655000

出　　处：《昆明医学院学报》2012年第7期91-93,共3页Journal of Kunming Medical College

基　　金：云南省应用基础研究基金资助项目(2009CD085)

摘　　要：目的建立高效液相色谱法(HPLC)间接测定肿瘤患者外周血二氢尿嘧啶脱氢酶(DPD)活性的方法.方法采用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.色谱柱:Xterra RPC18(250 mm×4.6 mm,5μm);流动相:甲醇-0.02 mol/L磷酸二氢钠(pH4.0),比例为5:95;流速为1.0 mL/min,H2U和U检测波长分别为204 nm、260 nm.结果 H2U线性范围为62.5～2 000μg/L,线性方程为:Y=7.48×10-4X+1.24×10-1,r=0.999 7(n=6),批内RSD小于7.7%,批间RSD小于13.8%,方法学回收率为99.63%～103.02%;U线性范围为15.625～500μg/L,线性方程为:Y=3.42×10-3X+2.58×10-2,r=0.999 3(n=6),批内RSD小于8.3%,批间RSD小于13.4%,方法学回收率为98.38%～109.08%.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 an HPLC method for determining dihydropyrimidine dehydrogenase activity in blood of cancer patients. Methods We determined the ratio of dihydrouracil （H2U） and uracil （U） in blood of patients with cancer by HPLC, and predicted the activity of dihydropyrimidine dehydrogenase （DPD） indirectly. The liquid chromatography column was Xterra RPC18 （250 mm x 4.6 mm, 5 I^m） ; the mobile phase was composed of 0.02 mol/L phosphate buffer （pH 4.0） and methanol （95：5） with a flow rate of 1.0 mL/min. The detection wavelength for H2U was 204 nm, and for U was 260 nm. Results A linearity was obtained from 62.5 to 2 000 I^g/L for H2U. The calibration equation of H2U was Y = 7.48 x 10-4X＋1.24 x 10-1, r = 0.999 7 （n = 6）. The RSD within day and between days were less than 7.7 % and 13.8 % for H2U. The recoveries of H2U were 99.63% ~ 103.02%. A linearity was obtained from 15.625 to 500 Ixg/L for U. The calibration equation of U was Y = 3.42 x 10-3X＋2.58 x 10-2, r= 0.9993 （n = 6）. The RSD within day and between days were less than 8.3% and 13.4% for U. The recoveries of U were 98.38% - 109.08%. Conclusion This method can be used for routine clinical determination of DPD activity in 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：R446.112[医药卫生—诊断学]