【作者】 高守红； 缪海均； 陈万生；

【Author】 Gao Shouhong,Miao Haijun,Chen Wansheng (Department of Pharmacy,Changzheng Hospital,Second Military Medical University,Shanghai 200003,PR China)

【机构】 第二军医大学长征医院药学部；

【摘要】 目的:建立一种简单、快速和灵敏的LC-MS/MS方法测定人血浆中丙戊酸和其三个代谢产物(3-OH-丙戊酸,4-ene-丙戊酸and5-OH-丙戊酸浓度);方法:采用苯甲酸为内标,血浆样品经固相萃取后,在Agilent Zorbax SB-C8(3.5μm,2.1×100mm,I.D.)色谱柱上,流动相为甲醇-10mM醋酸铵(内含0.1%甲酸)(80∶20),流速0.3mL/min,整个色谱方法运行2min,在Agilent G6410A串联质谱用负离子检测模式;结果:丙戊酸、3-OH-丙戊酸,4-ene-丙戊酸和5-OH-丙戊酸最低定量限分别为2030,51.5,50.15和51.25ng/mL;丙戊酸和其三个代谢产物的r>0.995,日内和日间精密度均<15.0%;结论:该方法成功的用于癫痫病人口服丙戊酸后测定丙戊酸和其三个代谢产物的定量研究。更多还原

【Abstract】 Objective:A rapid and sensitive method using liquid chromatography-tandem mass spectroscopy (LC-MS/MS) was developed and validated for simultaneous quantitative determination of valproic acid and three major metabolites (3-OH-valproic acid,4-ene-valproic acid and 5-OH-valproic acid) in human plasma.Methods:The analytes and internal standard were isolated from 200μL samples by solid phase extraction using a ZORBAX SB-C 8 column (3.5μm,2.1×100 mm) with an isocratic mobile phase consisting of methanol-10mM ammonium acetate (80:20,v/v) containing 0.1% formic acid at a flow rate of 0.3 mL/min.The method had a chromatographic total run time of 2.0min.The Agilent G6410A triple quadrupole LC/MS system was operated under a multiple reaction monitoring and single ion monitoring mode using the electrospray ionization technique in a negative mode.Results:The lower limit of quantification of valproic acid,3-OH-valproic acid,4-ene-valproic acid and 5-OH-valproic acid of the method was 2030,51.5,50.15 and 51.25 ng/mL,respectively.The method was linear for valproic acid and the three metabolites with correlation coefficients>0.995 for all analytes.The intra-day and inter-day accuracy and precision of the assay were less than 15.0%.Conclusions:This analytical method was successfully used to assay plasma concentrations of valproic acid and the three metabolites in human plasma from epileptic patients.更多还原