HPLC-MS/MS法测定人血浆中卡马西平浓度

Establishment of Concentration Determination Method of Carbamazepine in Human Plasma by HPLC-MS/MS

目的采用高效液相色谱-串联质谱法（HPLC-MS/MS）建立人血浆中卡马西平浓度测定方法,应用于药代动力学实验室生物样品检测室间质量评价。方法采用HPLC-MS/MS方法,以卡马西平-D8对照品为内标,Agilent Eclipse plus C18（4.6 mm×100 mm,3.5μm）为分析柱,流动相为含5 mmol/L甲酸铵的甲醇-水体系（90∶10,v∶v）,流速为0.5 m L/min,采用多重反应监测（MRM）模式进行检测。结果卡马西平的线性范围为5-1 000 ng/m L（r2=0.998 9）,定量下限为5 ng/m L,日内、日间精密度和准确度,绝对回收率和基质效应以及样品稳定性均符合生物样品测定需要,室间质评血样结果通过质量评价标准验证。结论本方法灵敏度高、特异性好,可用于人血浆中卡马西平的浓度测定以及药代动力学实验室生物样品检测室间质量评价。

OBJECTIVE To establish an HPLC-MS / MS method for determination of carbamazepine in human plasma and applied to the quality evaluation of pharmacokinetic laboratory biological sample. METHODS HPLC-MS / MS method was established to determine carbamazepine in human plasma,and carbamazepine-D8 was employed as the internal standard. The analyses were separated on a Agilent Eclipse plus C18（ 4.6 mm×100 mm,3.5 μm） as the mobile phase consisting of methanol-water（ 90 ∶10,v ∶v） containing 5mmol / L ammonium format,at a flow rate of 0.5 m L / min. The mass spectrometer was operated in multiple reactions monitoring（ MRM）mode.RESULTS The linearity range of carbamazepine was 5-1 000 ng / m L（ r2= 0.998 9） with a sensitivity of 5 ng / m L as the lower limit of the quantification. The intra-day and inter-day precisions,the recoveries and matrix effects at three spiked levels,and the sample stability were all suitable to the biological samples determination. The blood results of interstitial room quality evaluation could pass the validation for quality evaluation criteria. CONCLUSION The established method is highly sensitive, specific for determination of carbamazepine in human plasma,and it can be used for the quality evaluation of pharmacokinetic laboratory biological sample measure.