内源性尿嘧啶和二氢尿嘧啶的简易高效液相色谱测定法  

A Simplified HPLC Method for Determination of Endogenous Uracil and Dihydrouracil

作　　者：黄红兵[1] 黄民[2] 李苏[3] 廖海[3] 刘韬[1] 余更生[3] 姜文奇[3] 

机构地区：[1]中山医科大学肿瘤医院药剂科,广州510060 [2]中山医科大学临床药理教研室,广州510080 [3]中山医科大学肿瘤医院内科,广州510060

出　　处：《中国临床药理学杂志》2001年第4期290-293,共4页The Chinese Journal of Clinical Pharmacology

摘　　要：目的：建立测定内源性嘧啶类物质尿嘧啶（Ｕ）及二氢尿嘧啶（ＵＨ２）的反相高效液相色谱法，并应用于癌症病人５－氟尿嘧啶（５－Ｆｕ）化疗前的嘧啶代谢缺陷及毒性预测。方法：色谱柱：大连依利特ＯＤＳ２（２５０ｍｍ×４．６ｍｍ， ５μｍ）。以５－Ｆｕ作内标，经沉淀蛋白并以异丙醇与乙酸乙酯混合液提取，进行色谱分析。流动相为０．０１ｍｏｌ·Ｌ^（－１）ＫＨ２ＰＯ４－Ｈ３ＰＯ４缓冲液（ｐＨ＝３．０），流速０．５ｍｌ·ｍｉｎ^（－１），检测波长２０４ｎｍ。结果： ＵＨ２的线性方程为Ｙ＝１３６．７０Ｘ－９．４２（ｒ＝０．９９０９， ｎ＝７）， Ｕ的线性方程为Ｙ＝ ７８．０５Ｘ＋１．０３（ｒ＝０．９９９３，ｎ＝７），线性浓度范围为８μｇ·Ｌ^（－１）～５００μｇ·Ｌ^（－１），ＵＨ２最低检测浓度为６μｇ·Ｌ^（－１），Ｕ最低检测浓度为４μｇ·Ｌ^（－１）。平均回收率ＵＨ２为６６．６％，Ｕ为７０．４％。结论：本方法在同一色谱条件下完全分离二种痕量内源性嘧啶类物质，并予以定量分析，结果可作为确定行５－氟尿嘧啶化疗方案的肿瘤病人个体化给药剂量的依据。OBJECTIVE: To establish a rapid and simplified method for determining endogenous uracil (U) and dihydrouracil (UH2) in human plasma. METHOD: A HP HPLC system equipped with a C18 column (250mm × 4.6mm, 5μm) was used in the assay of U and UH2.The plasma sample was extracted with a solution of 15% of isopropanol in ethylacetate. 5-Fluorouracil (5-Fu) was chosen as the internal standard. The mobile phase was composed of a phosphate buffer solution (100 mmol.L^(-1), pH=3.0) and UV detection wavelength was 204 um. RESULT: The calibration curve equations of UH2 and U were Y=103.7X - 9.42 (r=0.9909, n=7) and Y=78.05X + 1 .03 (r=0.9993, n=7), respectively. The linear range was 8-500 μg.L^(-1) for both UH2 and U. The limitation of quantification was 6μg.L^(-1) for UH2 and 4μg.L^(-1) for U. CONCLUTION: This reversed-phase HPLC method is simple, rapid and sensitive. It can be used to detect the dihydropyrimidine dehydrogenase activity in human plasma.

关 键 词：内源性尿嘧啶 二氢尿嘧啶 高效液相色谱法 

分 类 号：R979.1[医药卫生—药品] R927[医药卫生—药学]