A validated liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry method for quantitation of cocaine and benzoylecgonine in human plasma
A validated liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry method for quantitation of cocaine and benzoylecgonine in human plasma
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DOI：
10.1093/jat/25.7.497
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发表时间：
2001-10-01
影响因子：
2.5
通讯作者：
Foltz, RL
中科院分区：
文献类型：
--
作者：
Lin, SN;Moody, DE;Foltz, RL
为了支持对治疗可卡因滥用的各种药物方案的研究，开发了一种准确、精确和敏感（2.5至750 ng/mL）的液相色谱-串联质谱分析方法，以测定人血浆中的可卡因和苯甲酰卵泡碱。加入可卡因-d3和苯甲酰卵磷脂-d3作为内标，固相萃取。在2.5 ng/mL浓度下，可卡因回收率为94.4%，苯甲酰茶碱回收率为80.3%。选择母体离子atm/z304和290进行反应监测，分别对可卡因和苯甲酰茶碱产生强片段atm/z182和168。该方法得到了充分的验证。定量下限为2.5 ng/mL时的平均测定浓度在目标的10.8%以内，在低（5 ng/mL）、中（50 ng/mL）和高（650 ng/mL）质量控制下测定的精密度范围为0.9 ~ 6.2% CV。经1% NaF处理的血浆中可卡因和苯甲酰ecgonine浓度在室温下保持7小时变化小于10%，在三次冻融循环中无显著变化。在- 20°C下保存11个月的血浆样品中，可卡因和苯甲酰冈碱的浓度保持稳定。两种分析物的甲醇原液是稳定的，在- 20°C下储存235天，保持在新制备的原液的2%以内。两种提取的分析物在甲醇溶液中重构，无论在- 20°C或室温下保存在自动进样器上，都可以稳定长达7天。该方法坚固、快速、可靠，已应用于700多个样品的药代动力学分析，以评估静脉注射（i.v）可卡因挑战和治疗药物之间的潜在相互作用。其中三名受试者接受40毫克（静脉注射）可卡因的结果证明了该方法的实用性。
In order to support studies on various medication protocols for the treatment of cocaine abuse, an accurate, precise, and sensitive (2.5 to 750 ng/mL) liquid chromatography-tandem mass spectrometry assay was developed to determine cocaine and benzoylecgonine in human plasma. Cocaine-d3and benzoylecgonine-d3were added as internal standards and samples were subjected to solid-phase extraction. Cocaine recovery was 94.4% and benzoylecgonine was 80.3% at 2.5 ng/mL. The selected reaction monitoring of parent ions atm/z304 and 290 resulted in strong fragments atm/z182 and 168 for cocaine and benzoylecgonine, respectively. The method was fully validated. The mean measured concentration at the 2.5 ng/mL, the lower limit of quantitation, was within 10.8 % of the target and the precision determined at the low (5 ng/mL), medium (50 ng/mL), and high (650 ng/mL) quality controls ranged from 0.9 to 6.2 %CV. Cocaine and benzoylecgonine concentrations in plasma treated with 1% NaF showed changes of less than 10% when maintained at room temperature for up to 7 h and no significant changes when subjected to three freeze-thaw cycles. The concentrations of cocaine and benzoylecgonine remained stable in plasma samples stored at −20°C for up to 11 months. Methanolic stock solutions of both analytes are stable, staying within 2% of the freshly prepared stock solutions, when stored at −20°C for up to 235 days. Both extracted analytes reconstituted in methanolic solutions are stable for up to seven days whether stored at −20°C or at room temperature on the autosampler. The method is rugged, rapid, and robust and has been applied to the batch analysis of more than 700 samples during pharmacokinetic profiling to assess potential interactions between intravenous (i.v.) cocaine challenge and treament medications. Results from three of these subjects receiving 40 mg (i.v.) cocaine demonstrate the utility of the method.