Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, SE-58758, Linköping, Sweden.
Drug Test Anal. 2014 Jun;6 Suppl 1:22-9. doi: 10.1002/dta.1637.
A sensitive and robust ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for quantification of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine and 3,4-methylenedioxy methamphetamine in hair samples. Segmented hair (10 mg) was incubated in 2M sodium hydroxide (80°C, 10 min) before liquid-liquid extraction with isooctane followed by centrifugation and evaporation of the organic phase to dryness. The residue was reconstituted in methanol:formate buffer pH 3 (20:80). The total run time was 4 min and after optimization of UHPLC-MS/MS-parameters validation included selectivity, matrix effects, recovery, process efficiency, calibration model and range, lower limit of quantification, precision and bias. The calibration curve ranged from 0.02 to 12.5 ng/mg, and the recovery was between 62 and 83%. During validation the bias was less than ±7% and the imprecision was less than 5% for all analytes. In routine analysis, fortified control samples demonstrated an imprecision <13% and control samples made from authentic hair demonstrated an imprecision <26%. The method was applied to samples from a controlled study of amphetamine intake as well as forensic hair samples previously analyzed with an ultra high performance liquid chromatography time of flight mass spectrometry (UHPLC-TOF-MS) screening method. The proposed method was suitable for quantification of these drugs in forensic cases including violent crimes, autopsy cases, drug testing and re-granting of driving licences. This study also demonstrated that if hair samples are divided into several short segments, the time point for intake of a small dose of amphetamine can be estimated, which might be useful when drug facilitated crimes are investigated.
建立并验证了一种灵敏、稳健的超高效液相色谱-串联质谱（UHPLC-MS/MS）法，用于检测头发样品中的苯丙胺、甲基苯丙胺、3,4-亚甲二氧基苯丙胺和 3,4-亚甲二氧基甲基苯丙胺。分段头发（10mg）在 2M 氢氧化钠（80°C，10min）中孵育，然后用异辛烷进行液液萃取，离心后将有机相蒸发至干。残渣用甲醇：甲酸缓冲液 pH 3（20:80）重新溶解。总运行时间为 4min，在优化 UHPLC-MS/MS 参数后进行验证，包括选择性、基质效应、回收率、过程效率、校准模型和范围、定量下限、精密度和偏差。校准曲线范围为 0.02-12.5ng/mg，回收率在 62-83%之间。在验证过程中，所有分析物的偏差均小于±7%，精密度均小于 5%。在常规分析中，加标对照样品的精密度<13%，真实头发对照样品的精密度<26%。该方法应用于苯丙胺摄入对照研究的样品以及先前用超高效液相色谱飞行时间质谱（UHPLC-TOF-MS）筛选方法分析的法医头发样品。该方法适用于法医案例中这些药物的定量分析，包括暴力犯罪、尸检案例、药物检测和驾驶执照重新发放。这项研究还表明，如果将头发样本分成几个短段，就可以估计摄入小剂量苯丙胺的时间点，这在调查药物助长的犯罪时可能很有用。
