Institute of Forensic Sciences, Forensic Toxicology Service, Faculty of Medicine, 15782-Santiago de Compostela, Spain.
J Appl Toxicol. 2013 Aug;33(8):740-5. doi: 10.1002/jat.2722. Epub 2012 Feb 15.
A simple method is presented for the simultaneous determination of morphine, 6-acetylmorphine, codeine, cocaine, benzoylecgonine, cocaethylene, methadone and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in vitreous humor by high-performance liquid chromatography with photodiode array detector after solid-phase extraction with Oasis® HLB cartridges and dichloromethane as eluent. The chromatographic process was carried out using an XTerra® RP8 column (250 × 4.6 mm i.d., 5 µm particle size) and a mobile phase composed of acetonitrile and pH 6.5 phosphate buffer in gradient mode. A linear response from the detector was obtained within the concentration range of 0.1-4 µg ml(-1) , with correlation coefficients higher than 0.99. The limits of detection were lower than 30 ng ml(-1) for all the drugs studied, the coefficients of variation fluctuated between 0.1 and 12.4%, and the average recoveries were higher than 78% for all the drugs except for EDDP, with a value of 66.4%. Finally, the proposed method was applied to 15 vitreous humor samples coming from individuals who had died from opiate and/or cocaine overdose, showing consumption of cocaine in 14 cases, methadone in five cases and heroin in three cases. Average concentrations of 0.30 µg ml(-1) for morphine, 0.24 µg ml(-1) for 6-acetylmorphine, 0.10 µg ml(-1) for codeine, 0.81 µg ml(-1) for cocaine, 1.26 µg ml(-1) for benzoylecgonine, 0.15 µg ml(-1) for cocaethylene, 0.11 µg ml(-1) for methadone and 0.68 µg ml(-1) for EDDP were obtained.
一种简单的方法被提出，用于高效液相色谱法与光电二极管阵列检测器同时测定玻璃体液中的吗啡、6-乙酰吗啡、可待因、可卡因、苯甲酰古柯碱、可乐定、美沙酮和 2-亚乙基-1,5-二甲基-3,3-二苯基吡咯烷（EDDP），经过固相萃取 Oasis®HLB 小柱和二氯甲烷作为洗脱液。色谱过程采用 XTerra®RP8 柱（250×4.6mm id，5μm 粒径）和由乙腈和 pH6.5 磷酸盐缓冲液组成的流动相，以梯度模式进行。检测器得到的线性响应范围为 0.1-4μgml(-1)，相关系数均高于 0.99。所有研究药物的检测限均低于 30ngml(-1)，变异系数在 0.1-12.4%之间波动，除 EDDP 外，所有药物的平均回收率均高于 78%，EDDP 的回收率为 66.4%。最后，该方法应用于 15 例来自阿片类药物和/或可卡因过量死亡个体的玻璃体液样本，结果显示 14 例存在可卡因消耗，5 例存在美沙酮消耗，3 例存在海洛因消耗。吗啡的平均浓度为 0.30μgml(-1)，6-乙酰吗啡为 0.24μgml(-1)，可待因为 0.10μgml(-1)，可卡因为 0.81μgml(-1)，苯甲酰古柯碱为 1.26μgml(-1)，可乐定 0.15μgml(-1)，美沙酮 0.11μgml(-1)，EDDP 为 0.68μgml(-1)。
