摘要
目的 建立同时测定人血浆中卡马西平、拉莫三嗪、氯硝西泮、地西泮及其代谢物奥沙西泮浓度的方法。方法 采用超高效液相色谱-质谱联用法（UPLC-MS/MS）,以磺胺甲噁唑（SMZ）为内标,血浆经甲醇直接沉淀后进样分析。色谱柱为WatersACQUITYUPLCHSSPFP柱（2.1mm×100mm,1.8μm）,流动相为0.1%甲酸的5mmol·L1乙酸铵水溶液-0.1%甲酸的甲醇溶液（0~5min,35∶65→10∶90）,流速为0.2mL·min1。电喷雾离子源,正离子多反应监测扫描分析,卡马西平、拉莫三嗪、氯硝西泮、地西泮和奥沙西泮的离子对分别为m/z237.0→194.06、m/z255.98→144.95、m/z316.01→270.0、m/z285.04→193.07和m/z287.02→241;内标磺胺甲噁唑的离子对为m/z253.96→91.97。结果 卡马西平、拉莫三嗪、氯硝西泮、地西泮和奥沙西泮血药浓度分别在2.4~600ng·mL1（r=0.9997）,2.52~630ng·mL1（r=0.9920）,2.08~520ng·mL1（r=0.9979）,2.28~570ng·mL1（r=0.9982）,8.0~800ng·mL1（r=0.9992）线性关系良好;最低检出限分别为0.24,0.63,0.52,0.57,3.2ng·mL1。日内、日间精密度均〈15%;提取回收率均〉70%,且RSD〈15%。结论 该方法灵敏、快速、专属性强,可用于临床血药浓度测定及药动学研究。
OBJECTIVE To develop the method for concentration determination of carbamazepine, lamotrigine, clonazepam, diazepam and oxazepam in human plasma. METHODS UPLC-MS/MS was adopted to analyze plasma with protein precipitated by methanol and sulfamethlazole（SMZ） was used as internal standard. Plasma samples were separated on Waters ACQUITY UPLC HSS PFP（2.1mm×100 mm, 1.8 μm） column with aqueous solution（0.1% formic acid 5 mmol.L^-1 ammonium acetate buffer）-0.1% formic acid method（0-5 min, 35 ： 65→10 ： 90） as mobile phase, and at a flow rate of 0.2 mL·min^-1. The protonated ion of samples was detected in positive ionization by multiple reaction monitoring（MRM） mode. The target compounds carbamazepine, lamotrigine, clonazepam, diazepam, oxazepam and SMZ were quantified with m/z 237.0 →194.06, m/z 255.98→144.95, m/z 316.01→270.0, m/z 285.04→193.07, m/z 287.02→241 and m/z 253.96→91.97, respectively. RESULTS The liner calibration curve of carbamazepine, lamotrigine, clonazepam, diazepam and oxazepam were obtained in the concentration range of 2.4-600 ng.mL-1（r=0.999 7）, 2.52-630 ng.mL-1（r=0.992 0）, 2.08-520 ng-mL-l（r=0.997 9）, 2.28-570 ng·mL-1 （r=0.998 2） and 8.0-800 ng·mL-1（r=0.999 2）, respectively. The lowest detection limit were 0.24 ng.mL^-1, 0.63 ng.mL^-1, 0.52 ng.mL-1, 0.57 ng.mL-1 and 3.2 ng·mL^-1, respectively. The RSD of inter-day and intra-day were less than 15%. The relative recovery was more than 70%, and the RSD was less than 15%. CONCLUSION The method is accurate, sensitive and suitable for blood concentration monitoring and pharmacokinetic study of carbamazepine, lamotrigine, clonazepam, diazepam and oxazepam.
出处
《中国现代应用药学》
CAS
CSCD
2013年第11期1215-1219,共5页
Chinese Journal of Modern Applied Pharmacy
基金
民航总医院院级课题(2012018)
民航科技项目(MHRD201235)
