论文部分内容阅读
目的:建立同时检测人血浆中卡马西平和苯巴比妥浓度的UPLC-MS/MS方法,应用于临床卡马西平和苯巴比妥血药浓度监测。方法:以地西泮为内标,血浆样品经乙腈沉淀蛋白,运用Waters XEVO TQD三重四级杆液质联用仪检测,色谱柱为ACQUITY UPLC~■ BEH C_(18)柱(50 mm×2.1 mm,1.7μm);流动相为乙腈(10 mmol·L~(~(-1))甲酸铵)-水(10 mmol·L~(~(-1))甲酸铵,含0.1%甲酸),梯度洗脱,流速为0.4 ml·min~(-1),柱温40℃;采用电喷雾离子化源(ESI),正离子多反应监测模式扫描分析;加入200μl乙腈沉淀蛋白,12 000×g离心10 min转移至1.5 ml EP管中取2μl上样。结果:卡马西平的保留时间为1.23 min,线性范围为0.25~25μg·ml~(-1)(r=0.999 7),最低定量限为0.01μg·ml~(~(-1));苯巴比妥的保留时间为1.11 min,线性范围为0.5~50μg·ml~(-1)(r=0.999 6),最低定量限为0.05μg·ml~(-1)。卡马西平的高、中、低浓度的回收率为(82.1±6.83)%,(82.91±4.3)%和(84.35±3.09)%;苯巴比妥的高、中、低浓度的回收率为(84.27±6.91)%,(84.32±7.74)%和(89.07±6.24)%。卡马西平和苯巴比妥的日内、日间RSD均<10%,血浆样品体系中的其他内源性物质不干扰测定。结论:该方法准确可靠,操作简便,重复性好,适于检测人血浆中卡马西平和苯巴比妥的血药浓度测定。
OBJECTIVE: To establish a simultaneous UPLC-MS / MS method for the simultaneous determination of carbamazepine and phenobarbital in human plasma, which is used in the clinical monitoring of carbamazepine and phenobarbital concentrations. Methods: The diazepam was used as an internal standard. The plasma samples were separated by acetonitrile precipitation and detected by Waters XEVO TQD Triple Quadrupole LC / MS. The chromatographic column was ACQUITY UPLC-BEH C 18 column (50 mm × 2.1 The mobile phase consisted of 10 mmol·L ~ (-1) ammonium formate) - water (10 mmol·L ~ (-1)) ammonium formate containing 0.1% formic acid) Gradient elution at a flow rate of 0.4 ml · min -1 with a column temperature of 40 ° C. Electrospray ionization (ESI) was used to scan and analyze the positive ion multi-reaction monitoring mode. 200 μl of acetonitrile precipitated protein was added and 12 000 × g Centrifuge 10 min transferred to 1.5 ml EP tube to take 2μl sample. Results: The retention time of carbamazepine was 1.23 min, the linear range was 0.25 ~ 25μg · ml -1 (r = 0.999 7), the limit of quantification was 0.01μg · ml ~ (-1) The retention time of barbital was 1.11 min and the linear range was 0.5-50 μg · ml -1 (r = 0.999 6). The lowest limit of quantification was 0.05 μg · ml -1. The recoveries of high, medium and low concentrations of carbamazepine were (82.1 ± 6.83)%, (82.91 ± 4.3)% and (84.35 ± 3.09)%, respectively. The recoveries of high, (84.27 ± 6.91)%, (84.32 ± 7.74)% and (89.07 ± 6.24)%, respectively. Carbamazepine and phenobarbital intraday, daytime RSD were <10%, plasma samples of other endogenous substances do not interfere with the determination. Conclusion: The method is accurate and reliable, simple and convenient to operate, reproducible and suitable for the determination of plasma concentrations of carbamazepine and phenobarbital in human plasma.