Gunnar Teemu, Eskola Tanja, Lillsunde Pirjo
National Public Health Institute, Drug Research Unit, Mannerheimintie 166, 00300 Helsinki, Finland.
Rapid Commun Mass Spectrom. 2006;20(4):673-9. doi: 10.1002/rcm.2356.
A toxicological analysis was developed and validated for simultaneous screening and quantification of methadone (METH) and its primary metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP). The method employs microscale liquid-liquid extraction (microLLE) and direct injection of a separated aliquot of the organic layer into a gas chromatography/mass spectrometric (GC/MS) system without any other pre-treatment stages. A fast GC/MS runtime (total 5.8 min; METH, Rt = 3.55 min; EDDP, Rt = 3.40 min) combined with rapid sample preparation allowed cost-efficient and routinely applicable performance with a low amount of manual work. The validated parameters included: linearity (25-1000 ng mL(-1) both; R(METH)2 = 0.998 and R(EDDP)2 = 0.997), accuracy (Bias(METH): from -0.05 to 11.3%, Bias(EDDP): from 1.11 to 4.37%); intra and inter-assay precision (RSD(METH): from 2.4 to 3.9%, from 4.89 to 10.3%; RSD(EDDP): from 4.50 to 6.20%, from 4.57 to 15.2%), extraction efficiency (METH = 95.5%; EDDP = 90.6%), LOQ(Meth,EDDP) = 25 ng mL(-1). Samples were stable for at least 25 h and no selectivity problems or baseline interference were observed. The method should be applicable for identifying and quantitative confirmation of possible misuse and/or illegal use of METH in toxicological cases.
开发并验证了一种毒理学分析方法，用于同时筛查和定量美沙酮（METH）及其主要代谢物2-亚乙基-1,5-二甲基-3,3-二苯基吡咯烷（EDDP）。该方法采用微量液-液萃取（microLLE），并将有机层的分离等分试样直接注入气相色谱/质谱（GC/MS）系统，无需任何其他预处理步骤。快速的GC/MS运行时间（总计5.8分钟；METH，保留时间 = 3.55分钟；EDDP，保留时间 = 3.40分钟）与快速的样品制备相结合，实现了低成本且常规适用的性能，人工操作量少。验证的参数包括：线性（两者均为25 - 1000 ng mL⁻¹；R(METH)² = 0.998，R(EDDP)² = 0.997）、准确度（偏差(METH)：-0.05%至11.3%，偏差(EDDP)：1.11%至4.37%）；批内和批间精密度（RSD(METH)：2.4%至3.9%，4.89%至10.3%；RSD(EDDP)：4.50%至6.20%，4.57%至15.2%）、萃取效率（METH = 95.5%；EDDP = 90.6%）、定量限(Meth,EDDP) = 25 ng mL⁻¹。样品至少在25小时内稳定，未观察到选择性问题或基线干扰。该方法应适用于在毒理学案例中识别和定量确认METH可能的滥用和/或非法使用情况。
