A high-throughput UHPLC-MS/MS method for the most frequently found compounds; tetrahydrocannabinol (THC), amphetamine, methamphetamine, MDMA, clonazepam, diazepam, nordiazepam, oxazepam, alprazolam, nitrazepam, morphine, and codeine, in driving under the influence of drugs (DUID) cases in whole blood, is presented. Automated sample preparation by 96-well supported liquid extraction (SLE) plates with ethyl acetate + heptane (80 + 20, v/v) as organic solvent was carried out on a Freedom Evo 200 platform from Tecan. An aliquot of 100 μL whole blood was used. Sample preparation time for 96 samples was 1.5 h. Compounds were separated with gradient elution on a C₁₈ column (50 × 2.1 mm, 1.7 μm) with a mobile phase consisting of 5 mM pH 10.2 ammonium formate and methanol. The run time was 4.5 min and 1 μL was injected on an Acquity UPLC I-Class system with a Xevo TQS tandem-quadrupole mass spectrometer in multiple-reaction monitoring mode (MRM) from Waters. Isotope labelled, ¹³C, internal standards (ISs) were used for all compounds except for alprazolam and morphine, which had deuterated analogs. Quantification was carried out with calibrators without whole blood matrix. Full validation was carried out according to international guidelines, and a new approach for evaluation of process efficiency (PE) has been presented. Linear or quadratic weighted (1/x) calibration curves were used with R² ≥ 0.999. The method showed satisfactory deviations ±16% when compared to the existing methods, and satisfactory agreement with proficiency testing control samples (z-score −1.6 to 1.8, n = 16 samples). The precision, estimated as the relative standard deviation (RSD) of the concentration difference between results from two independent analyses of authentic whole blood samples, was ≤7.2% in antemortem and ≤9.3% in postmortem samples. Recovery was ≥85% for all the compounds, except morphine ≥62% and THC ≥ 50%. PE was satisfactory for all the compounds with low variation in IS response, RSD ≤ 16% (THC 27%) in antemortem samples and ≤34% (THC 66%) in postmortem samples. To the best of our knowledge, this is the first automated 96-well SLE UHPLC-MS/MS method developed for the simultaneous determination of these 12 compounds in whole blood covering the concentration ranges found in forensic samples. The method has been used in routine work during the last ten months, analysing about 9900 antemortem and 1000 postmortem whole blood samples, and has proven to be robust and reliable.

针对最常发现的化合物的高通量UHPLC-MS / MS方法；介绍了在全血中药物（DUID）的影响下驾驶的四氢大麻酚（THC），苯丙胺，甲基苯丙胺，摇头丸，氯硝西am，地西epa，诺地西p，奥沙西epa，阿普唑仑，硝西epa，吗啡和可待因。在来自Tecan的Freedom Evo 200平台上，使用乙酸乙酯+庚烷（80 + 20，v / v）作为有机溶剂，通过96孔支撑液体萃取（SLE）板进行自动样品制备。使用100μL全血的等分试样。96个样品的样品制备时间为1.5 h。在C ₁₈上通过梯度洗脱分离化合物色谱柱（50×2.1 mm，1.7μm），流动相由5 mM pH 10.2甲酸铵和甲醇组成。运行时间为4.5分钟，并在沃特世（Waters）的多反应监测模式（MRM）中使用Xevo TQS串联四极杆质谱仪在Acquity UPLC I-Class系统上注入1μL。所有化合物均使用同位素标记的¹³ C内标（IS），除阿普唑仑和吗啡外，它们均具有氘代类似物。使用没有全血基质的校准物进行定量。根据国际准则进行了全面验证，并提出了一种评估过程效率（PE）的新方法。R ²使用线性或二次加权（1 / x）校准曲线 ≥0.999。与现有方法相比，该方法显示出令人满意的偏差±16％，并且与水平测试对照样品（z评分-1.6至1.8，n = 16个样品）具有令人满意的一致性。准确度估计为两次独立分析的真实全血样品结果之间的浓度差的相对标准偏差（RSD），在死前样品中≤7.2％，在死后样品中≤9.3％。除吗啡≥62％和THC≥50％外，所有化合物的回收率均≥85％。PE对所有IS反应变化低，死前样品的RSD≤16％（THC 27％）和死后样品的≤34％（THC 66％）的化合物均令人满意。据我们所知，这是第一个开发的用于同时测定法医样品中浓度范围的全血中这12种化合物的自动96孔SLE UHPLC-MS / MS方法。该方法已在过去的10个月中用于日常工作，分析了大约9900个死前和1000个事后全血样本，并被证明是可靠且可靠的。