液相色谱-质谱联用法测定癫痫患者血浆中丙戊酸及其代谢产物2-丙基-4-戊烯酸浓度  

Simultaneous determination of valproic acid and its metabolite 2-propyl-4-pentenoic acid in epileptic patients plasma by liquid chromatography-tandem mass spectrometry

作　　者：李力[1] 李贺[1] 余杨[1] 廉洪[1] LI Li;LI He;YU Yang;LIAN Hong(Department of Pharmacy, Zhejiang Hospital,Hangzhou 310013,China)

机构地区：[1]浙江医院药剂科,杭州310013

出　　处：《中国临床药理学杂志》2018年第23期2757-2760,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家科技重大专项项目-老年病新药临床技术平台基金资助项目(2013ZX09303005);浙江省自然科学基金资助项目(LQ15H310003)

摘　　要：目的建立液相色谱-质谱联用法同时测定丙戊酸(VPA)及其毒性代谢产物2-丙基-4-戊烯酸(4-ene-VPA)浓度。方法人血浆样品用乙腈直接沉淀蛋白后,色谱柱:Agilent Eclipse Plus-C_(18)(2. 1 mm×10. 0 mm,3. 5μm),流动相:乙腈-10 mmol·L^(-1)醋酸铵水溶液(均含0. 1%甲酸),梯度洗脱,用电喷雾离子化源,负离子方式,扫描方式为多反应监测(MRM),用于监测的离子反应分别为m/z 143. 2→143. 2(丙戊酸)、m/z 140. 8→140. 8(2-丙基-4-戊烯酸)和m/z 373. 0→329. 1(内标银杏酸(C17:1))。考察该方法的专属性、标准曲线与定量下限、精密度与回收率、基质效应和稳定性。结果 VPA和4-eneVPA分别在0. 50～200. 00μg·m L^(-1)和0. 05～12. 50μg·m L^(-1)内线性关系良好(r分别为0. 998 4和0. 997 5),定量下限分别为0. 50和0. 05μg·m L^(-1)批内、批间RSD均小于8. 59%,提取回收率分别为76. 45%～86. 78%和78. 44%～83. 46%,内标归一化基质效应因子分别为93. 67%～97. 82%和92. 04%～104. 35%。结论本方法简洁、灵敏、专属性好,适用于人血浆中同时测定VPA和4-eneVPA浓度,可用于丙戊酸钠的血药浓度监测。Objective To establish a specific and sensitive liquid chromatography - tandem mass spectrometry method for simultaneous determination of valproic acid (VPA) and its toxic metabolite 2-propyl-4 - pentenoic acid (4-ene-VPA) in epileptic patients plasma,and apply this method for therapeutic drug monitoring.Methods The plasma sample was treated by one-step protein precipitation with acetonitrile.Chromatographic separation was carried out on Agilent Eclipse Plus-C18 (2.1 mm × 10.0 mm,3.5 μm) and eluted with linear gradient using acetonitrile and 10 mmol·L^-1 ammonium acetate,both of which were acidified with 0.1% formic acid.Detection of the analytes were achieved using negative ion electrospray ionization (ESI) in the multiple reaction monitoring (MRM) mode.The MS /MS ion transitions monitored were m/z 143.2→143.2 (VPA) ,m/z 140.8→140.8 (4-ene-VPA) and m/z 373.0→329.1(Ginkgolic acid(C17: 1)) .The specificity,standard curve and lower limitation of quantitation,precision and recovery rate,stability and matrix effect were investigated.Results The linear calibration curves for VPA and 4-ene-VPA were demonstrated in the concentration ranges of 0.50-200.00 μg·mL^-1 (r = 0.998 4) and 0.05-12.50 μg·mL^-1 (r = 0.997 5) separately.The lower limit of quantification of VPA and 4-ene-VPA were 0.50 μg·mL^-1and 0.05 μg·mL^-1.The intra and inter assay precisions were both less than 8.59%,the accuracies were all in standard ranges The extraction recoveries VPA and 4-ene-VPA were between 76.45%-86.78%,78.44%-83.46%,the internal standard normalized matrix effect were 93.67%-97.82% and 92.04%-104.35% respectively.Conclusion The developed method is proved to be simple,sensitive,specific,and could be suitable for simultaneous determination of valproic acid and its toxic metabolite2-propyl-4-pentenoic acid in human plasma,the validation method can be used for the therapeutic drug monitoring of sodium valproate.

关 键 词：丙戊酸 2-丙基-4-戊烯酸 毒性代谢产物 液相色谱-质谱联用法 血药浓度监测 

分 类 号：R971[医药卫生—药品]