Objectives: To establish a method for the detection of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Methods: The blood samples were treated with 1-butyl-3-methylimidazolium hexafluorophosphate as an extraction solvent. The samples were extracted by ultrasound-assisted extraction and separated by ZORBAX Eclipse Plus C18, 95Å column. The mobile phase A aqueous solution containing 0.1% formic acid and 10 mmol/L ammonium acetate, and mobile phase B mixed organic solvent containing acetonitrile/methanol (Vacetonitrile∶Vmethanol=2∶3) were used for gradient elution at the flow rate of 1.00 mL/min. An electrospray ion source in positive mode was used for detection in the multiple reaction monitoring.
Results: The linearities of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples were good within the corresponding range, with correlation coefficients (r) greater than 0.995 6. The limits of detection were 3.00, 0.40 and 1.30 ng/mL, respectively. The limit of quantitation were 8.00, 1.00 and 5.00 ng/mL, respectively. The extraction recoveries ranged from 76.00% to 106.44%. The relative standard deviations of the intra-day and inter-day precisions were less than 16%. Carbamazepine and its main metabolite 10,11-dihydro-10,11-epoxycarbamazepine were detected in blood samples of death cases with a mass concentration of 2.71 μg/mL and 252.14 ng/mL, respectively.
Conclusions: This method has high sensitivity and good selectivity, which is suitable for the detection of carbamazepine and its metabolites in blood samples, and can be used for carbamazepine-related forensic identifications.
目的: 建立LC-MS/MS法测定血液样品中卡马西平及其代谢物10，11-二氢-10，11-环氧卡马西平和10，11-二氢-10-羟基卡马西平的方法。方法: 以1-丁基-3-甲基咪唑六氟磷酸盐离子液体为萃取剂处理血液样品，通过超声辅助萃取，采用ZORBAX Eclipse Plus C18，95Å色谱柱分离，流动相A为含0.1%甲酸、10 mmol/L乙酸铵的水溶液，流动相B为混合有机溶剂（V乙腈∶V甲醇=2∶3），梯度洗脱，流速1.00 mL/min，采用电喷雾离子源、正离子模式，多反应监测模式进行检测。结果: 血液样品中卡马西平及其代谢物10，11-二氢-10，11-环氧卡马西平和10，11-二氢-10-羟基卡马西平在相应范围内线性关系良好，相关系数（r）均大于0.995 6，检出限分别为3.00、0.40和1.30 ng/mL，定量限分别为8.00、1.00和5.00 ng/mL，提取回收率为76.00%~106.44%，日内和日间精密度均小于16%。采用本方法从死亡案件的血液样品中检出卡马西平和主要代谢产物10，11-二氢-10，11-环氧卡马西平成分，质量浓度分别为2.71 μg/mL和252.14 ng/mL。结论: 本方法灵敏度高、选择性好，适用于血液样品中卡马西平及其代谢物的检测，可应用于卡马西平相关的法医学鉴定。.
Keywords: 10,11-dihydro-10,11-epoxycarbamazepine; 10,11-dihydro-10-hydroxycarbamazepine; LC-MS/MS; carbamazepine; forensic toxicological analysis; metabolites.