【摘要】 目的 用液相色谱-串联质谱法(LC-MS/MS)建立尿液基质中3种钩吻生物碱的检测方法，为钩吻中毒事件应急处置提供技术支持。方法 尿液经过固相萃取(SPE)净化后，用LC-MS/MS法的多反应监测模式(MRM)检测，并考察SPE净化的填料种类、上样体积和洗脱体积等对测定的影响。结果 使用WCX(弱阳离子交换)固相萃取柱的效果较好，上柱3.00 mL尿样时，以2 mL 5%氨水-甲醇溶液能使钩吻生物碱洗脱充分；经过验证，3种钩吻生物碱的检出限为0.01～0.02μg/L,定量限为0.02～0.05μg/L,在0.02～15.0μg/L范围内钩吻生物碱的浓度与峰面积呈良好的线性关系(r≥0.995)。以空白人尿液为基质，进行低、中、高3个浓度水平(0.05～5.00μg/L)的加标回收试验，钩吻生物碱的回收率为84.7%～105.0%,相对标准偏差(RSD)3.2%～6.4%(n=6)。结论 本法较好地满足了尿液基质中3种钩吻生物碱的定量测定，灵敏度较高，净化步骤较简便，能用于疑似钩吻中毒的快速检验。更多还原

【Abstract】 Objective The liquid chromatography-tandem mass spectrometry(LC-MS/MS) method was established for determination of three gelsemium elegans alkaloids in urine matrix, which can provide technical support for the emergency response to poisoning incident. Methods Urine was purified by solid-phase extraction(SPE) and then was determined using LC-MS/MS in the mode of multiple reaction monitoring(MRM). The effect of key factors of SPE purification, such as type of sorbents, sample volume, and elution volume, on the determination of gelsemium elegans alkaloids were investigated. Results The results determined by SPE using weak cation exchange sorbent(WCX) was better than two other sorbents, i.e., MCX and HLB. When 3.00 mL of urine was added to cartridge, 2 mL of 5% ammonia-methanol solution resulted in adequate elution of gelsemium elegans alkaloids. The limits of detection of the three alkaloids in gelsemium elegans were validated to be in the range of 0.01-0.02 μg/L, and the limits of quantification were in the range of 0.02-0.05 μg/L.The concentration of alkaloids in the range of 0.02-15.0 μg/L showed a good linear relationship with peak area(r ≥0.995). The spiked recovery tests were performed at three concentration levels(0.05-5.00 μg/L) of low, medium and high using blank human urine as the matrix. The recoveries of three alkaloids ranged from 84.7% to 105.0% with the relative standard deviation(RSD) of 3.2% to 6.4%(n=6). Conclusion The method meets the requirements for the quantitative determination of three gelsemium alkaloids in the urine matrix with higher sensitivity and simpler purification steps. It may be used for rapid test of suspected poisoning incidents caused by gelsmium elegans.更多还原