UPLC–MS/MS同时测定大鼠血、脑脊液中奎硫平及其活性代谢物

Simultaneous Determination of Quetiapine and Its Active Metabolites in Rat Plasma and Cerebrospinal Fluid by UPLC-MS/MS

目的建立同时测定大鼠血、脑脊液中奎硫平及其活性代谢物7-羟基奎硫平羟7-羟基-N-去烷基奎硫平浓度的UPLC-MS/MS法。方法样品经碱化后用叔丁基甲醚提取,采用Acquity UPLCTM BEH C18柱(2.1mm×50mm,1.7μm),柱温40℃,以水(含50mmol·L-1醋酸铵,3‰甲酸)-乙腈(73∶27)为流动相,流速0.25mL·min-1,进样量5μL。采用电喷雾离子化四极杆串联质谱,选择离子监测方式进行扫描,测定样品浓度,卡马西平作为内标。结果血浆中:奎硫平在0.0775～155μg·L-1,7-羟基奎硫平在0.049～98μg·L-1,7-羟基-N-去烷基奎硫平在0.067～670μg·L-1内线性关系良好,萃取回收率均>89%,方法回收率均>87%,,内羟,间,密度RSD均<9%。脑脊液中:奎硫平在0.02～38.75μg·L-1,7-羟基奎硫平在0.0098～29.5μg·L-1,7-羟基-N-去烷基奎硫平在0.067～670μg·L-1内线性关系良好,萃取回收率均>87%,方法回收率均>85%,,内羟,间,密度RSD均<16%。结论该方法该该度高、快速羟该确羟有效,且血、脑中浓度均可测定,为研究转运蛋白对抗,神分裂药物进入血脑屏障的影响提供了依据。

OBJECTIVE To establish a UPLC-MS/MS method for the simultaneous determination ofquetiapine and its active metabolites （7-hydroxy quetiapine and 7-hydroxy-N-dealkly quetiapine） in rat plasma and cerebrospinal fluid. METHODS The samples were extracted by Methyl t-Butyl Ether after alkalinization. The UPLC seperation of the compounds was performed on a Acquity UPLC^TM BEH C18 （2.1 mm×50 mm, 1.7μm） column. The mobile phase column, the temperature was at 40℃, consisted of water （ammonium acetate： 50 mmol·L^-1, formic acid： 3‰）- acetonitrile （73：27）. The flow rate was 0.25 mL·min^-1 and the sample size was 5 μL. Electrospray ionization （ESI） source was applied and operated in the positive ion mode. Carbamazepine was employed as the internal standard. RESULTS In plasma： The calibration curves were linear in the rang of 0.077 5-155 μg·L^-1 for quetiapine, 0.049-98 μg·L^-1 for 7-hydroxy quetiapine and 0.067-670 μg·L^-1 for 7-hydroxy-N-dealkly quetiapine, respectively. The average extraction recoveries for all the three samples were above 89%. The methodology recoveries were much higher than 87%. The intra-day and inter-day RSD were less than 9 %. In cerebrospinal fluid： The calibration curves were linear in the rang of 0.02-38.75 μg·L^-1 for quetiapine, 0.009 8-29.5 μg·L^-1 for 7-hydroxy quetiapine and 0.067-670 μg·L^-1 for 7-hydroxy-N-dealkly quetiapine, respectively. The average extraction recoveries for all the three samples were above 87%. The methodology recoveries were much higher than 85%. The intra-day and inter-day RSDs were less than 16%. CONCLUSION The method is highly sensitive, rapid, accurate and validated and it can be used in both plasma and cerebrospinal fluid obtained in the study of transport proteins＇ influence on the antipsychotics after passing the blood-brain barrier.