Koželj Gordana, Perharič Lucija, Stanovnik Lovro, Prosen Helena
Institute of Forensic Medicine, Faculty of Medicine, Korytkova 2, SI-1000 Ljubljana, Slovenia.
Department of Infectious Diseases and Environmental Risk, National Institute of Public Health, Trubarjeva 2, SI-1000 Ljubljana, Slovenia.
J Pharm Biomed Anal. 2014 Aug 5;96:197-206. doi: 10.1016/j.jpba.2014.03.037. Epub 2014 Apr 1.
A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of atropine and scopolamine in 100μL human plasma was developed and validated. Sample pretreatment consisted of protein precipitation with acetonitrile followed by a concentration step. Analytes and levobupivacaine (internal standard) were separated on a Zorbax XDB-CN column (75mm×4.6mm i.d., 3.5μm) with gradient elution (purified water, acetonitrile, formic acid). The triple quadrupole MS was operated in ESI positive mode. Matrix effect was estimated for deproteinised plasma samples. Selected reaction monitoring (SRM) was used for quantification in the range of 0.10-50.00ng/mL. Interday precision for both tropanes and intraday precision for atropine was <10%, intraday precision for scopolamine was <14% and <18% at lower limit of quantification (LLOQ). Mean interday and intraday accuracies for atropine were within ±7% and for scopolamine within ±11%. The method can be used for determination of therapeutic and toxic levels of both compounds and has been successfully applied to a study of pharmacodynamic and pharmacokinetic properties of tropanes, where plasma samples of volunteers were collected at fixed time intervals after ingestion of a buckwheat meal, spiked with five low doses of tropanes.
建立并验证了一种用于测定100μL人血浆中阿托品和东莨菪碱的液相色谱 - 串联质谱（LC-MS/MS）方法。样品预处理包括用乙腈进行蛋白沉淀，随后进行浓缩步骤。分析物和左旋布比卡因（内标）在Zorbax XDB-CN柱（75mm×4.6mm内径，3.5μm）上通过梯度洗脱（纯化水、乙腈、甲酸）进行分离。三重四极杆质谱在电喷雾电离正模式下运行。对脱蛋白血浆样品评估基质效应。采用选择反应监测（SRM）在0.10 - 50.00ng/mL范围内进行定量。两种托烷类药物的日间精密度以及阿托品的日内精密度均<10%，东莨菪碱的日内精密度在定量下限（LLOQ）时<14%且<18%。阿托品的日间和日内平均准确度在±7%以内，东莨菪碱在±11%以内。该方法可用于测定两种化合物的治疗和中毒水平，并已成功应用于托烷类药物的药效学和药代动力学性质研究，其中在摄入掺有五种低剂量托烷类药物的荞麦餐后，按固定时间间隔采集志愿者的血浆样本。
