LC-MS法测定大鼠脑部海马区中苯巴比妥的含量  

Determination of phenobarbital in rat hippocampus by LC-MS

作　　者：杨志宏[1] 刘晓东[1] 谢林[1] 梁艳[1] 杨慧文[1] 

机构地区：[1]中国药科大学药物代谢与动力学研究中心,南京210009

出　　处：《中国药科大学学报》2006年第2期142-145,共4页Journal of China Pharmaceutical University

基　　金：国家高技术研究发展计划("八六三"计划)资助项目(No.2003AA2Z347A);江苏省药物代谢动力学重点实验室资助项目(No.BM2001201)~~

摘　　要：目的：建立大鼠脑部海马区中苯巴比妥的LC-MS测定方法。方法：海马匀浆液中加入内标茶碱。用乙醚／环己烷混合系统提取、挥干，用甲醇复溶，高速离心后，取上清液进行LC-MS测定。色谱柱为Shim-pack ODSC18（150mm×2．0mm ID，5．0μm），离子源为ESI，检测离子为[M-H]^-：苯巴比妥（m／z）：231；茶碱（m／z）：179，流动相A为超纯水＋0．03％甲酸＋0．004％三乙胺，流动相B为甲醇，以0．2mL／min的流速进行梯度洗脱。结果：此条件下苯巴比妥与内标茶碱显示良好分离；方法回收率大于87％；批内批间变异系数均小于10％；线性范围为10～2000ng／mL；可定量下限为10ng/mL。在相应的生物样品中标准曲线的相关系数r大于0．9997，质控样本、冻融试验及基质效应均符合要求。结论：该方法经考查符合生物样品测定要求，可用于海马中苯巴比妥的含量测定。Aim：To develop a sensitive and specific LC-MS method for the quantitative analysis of phenobarbital （PB） in rat hippocampus. Methods：PB was extracted from rat hippocampus by the mixed solvents of ether and cyclohexane after theophylline was added as internal standard. After evaporation, the residual was reconstituted with methanol and centrifuged. The upper aliquot was used for injection onto HPLC system. The chromatographic analyses were performed on a by Shim-pack ODS C18 column （150 mm× 2.0 mm 1D,5.0 μm） with the mobile phase which was operated in the gradient mode using mobile phase A （methanol-0.03% formic acid-0.004% triethylamine） and mobile phase B （methanol） at a flow rat of 0.2 mL/min. Detection was carried out on an electrospray ionization （ESI） mass spectrometer; operated in selected ion monitoring （SIM） and negative-ionization mode of target ion m/z 231 for PB and m/z 179 for the theophylline. Results： Under this analytic setting, there was good resolution between PB and IS. The calibration curve was linear in the range of 10～ 2000 ng/mL with r 〉 0.99 and the limit of quantitation of 10 ng/ mL. The recoveries of the analytes were more than 87%, and both intra-batch and inter-batch RSD were less than 10%. In addition, freeze-thaw stability,matrix effect, and quality control were with acceptable data. Conclusion： A sensitive, reliable and accurate method suitable for the quantitation of PB in rat hippocampus has been established.

关 键 词：苯巴比妥 LC-MS法 海马 含量测定 

分 类 号：R96[医药卫生—药理学]