高效液相色谱法测定血中伊立替康及活性代谢物SN-38浓度

Determination of Irinotecan and Its Active Metabolite SN-38 in Plasma by High Performance Liquid Chromatography Coupled with Fluorescence Detection

目的：建立高效液相色谱法同时测定结直肠癌患者血中的伊立替康（CPT-11）及其活性代谢物7-乙基-10-羟基喜树碱（SN-38）的浓度，并对我院基因型指导给药方案进行评价。方法：以2μg·mL-110-羟基喜树碱作为内标，先用100μL 10%高氯酸沉淀蛋白，再用50μL 10%高氯酸酸化血浆。采用Agilent ZORBAX Eclipse C8色谱柱（4.6 mm×150 mm，5μm）对CPT-11和SN-38进行分离；以0.05 mol·L^-1的磷酸二氢钠-乙腈-三乙胺（75∶25∶0.1，v∶v，磷酸调pH 3.0）为流动相；荧光检测波长：激发波长380 nm，发射波长550 nm。结果：人血浆中CPT-11和SN-38线性范围均为3-1000 ng·mL^-1，定量下限为3 ng·mL^-1；准确度分别是98.5%和100.0%；回收率分别是83.8%和84.3%。结论：本方法可靠、简便、快速，可为伊立替康个体化给药提供参考。

Objective: To determine irinotecan (CPT-11) and its active metabolite 7-ethyl-10-hydroxy-camptothecin (SN-38) in plasma of patients with colorectal cancer by high performance liquid chromatography coupled with fluorescence detection and to evaluate the dosage regimen by genotype information. Methods:Irinotecan and metabolites were detected in plasma. The internal standard was 2μg·mL-1 10-hydroxycamp-tothecin. Protein in plasma was precipitated with 10% perchloric acid before acidification with 10% perchloric acid. Separation of the compounds was achieved using an Agilent ZORBAX Eclipse C8 (4.6 mm×150 mm, 5μm) analytical column. The elution was performed with a mobile phase of 0.05 mol·L-1 sodium dihydrogen phosphate solution -acetonitrile (75∶25, v∶v) containing 0.1% triethylamine. The fluorescence detector excitation and emis-sion wavelengths were set at 380 and 550 nm, respectively. Results: The linear ranges for CPT-11 and SN-38 were 3-1000 ng·mL-1. The limits of quantification of CPT-11 and SN-38 were 3 ng·mL-1. The average accuracy of CPT-11 and SN-38 were 98.5% and 100.0%; the average recoveries were 83.8% and 84.3%. Conclusion: This method is reliable, convenient and efficient, which can provide reference for individualized drug administration.