Federal Public Service Justice, National Institute of Criminalistics and Criminology, Brussels, Belgium.
Ther Drug Monit. 2013 Aug;35(4):510-21. doi: 10.1097/FTD.0b013e31828e7e6b.
A sensitive and selective ultra performance liquid chromatographic-tandem mass spectrometric method was developed and fully validated for the simultaneous determination of (in order of chromatographic elution) methylecgonine, pholcodine, morphine, hydromorphone, oxymorphone, norcodeine, codeine, dihydrocodeine, oxycodone, 6-Monoacetylmorphine (6-MAM), hydrocodone, ethylmorphine, norfentanyl, benzoylecgonine, tramadol, normeperidine, meperidine, cocaine, pentazocine, cocaethylene, fentanyl, norbuprenorphine, 2-ethylidine-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), buprenorphine, propoxyphene, and methadone in blood. The matrixes analyzed during the validation experiments were as follows: citrated blank plasma for calibrators, fluoride blank plasma for internal quality control (QC), lyophilized serum for external QC, fluoride plasma and whole blood for authentic samples, and lyophilized serum and whole blood for proficiency testing schemes. Samples were extracted with cation exchange solid-phase extraction cartridges. The target drugs were separated and quantified in a chromatographic run of 8.1 minutes using 0.1% formic acid in water and methanol (with 0.1% formic acid) as mobile phase. The limit of quantification ranged from 0.5 to 2.5 ng/mL depending on the compound and the therapeutic concentration. The intra- and interassay precision was less than 15% for all the compounds (except for pentazocine and EDDP, which was <20%) determined with 2 internal and 2 external QC samples, and the bias was within ±15% (except for methylecgonine, which was <20%). Extraction efficiency was greater than 70% for all the compounds except for EDDP. Matrix effects were evaluated with authentic blood samples (n = 10), and they ranged from 47 to 95%, but they were compensated for most analytes using deuterated analogs as internal standards. Prepared samples were stable for 62 hours in the autosampler. This method was successfully applied to authentic samples (n = 120), involving the use of heroin, cocaine, tramadol, and methadone, and to proficiency testing schemes.
建立并充分验证了一种灵敏、选择性的超高效液相色谱-串联质谱法，用于同时测定（按色谱洗脱顺序）：甲基可卡因而、福可定、吗啡、氢吗啡酮、羟吗啡酮、去甲可待因、可待因、二氢可待因、羟考酮、6-单乙酰吗啡（6-MAM）、氢可酮、乙基吗啡、诺夫啡烷、苯甲酰可卡因而、曲马多、去甲哌替啶、哌替啶、可卡因、喷他佐辛、可乐定、芬太尼、去甲纳布啡、2-亚乙基-1,5-二甲基-3,3-二苯基吡咯烷（EDDP）、丁丙诺啡、丙氧芬和美沙酮。在验证实验中分析的基质如下：用于校准品的柠檬酸盐空白血浆、用于内部质控（QC）的氟化物空白血浆、用于外部 QC 的冻干血清、用于真实样品的氟化物血浆和全血，以及用于能力验证计划的冻干血清和全血。样品用阳离子交换固相萃取小柱提取。在色谱运行 8.1 分钟内，使用 0.1%甲酸在水中和甲醇（含 0.1%甲酸）作为流动相，分离和定量目标药物。根据化合物和治疗浓度的不同，定量下限范围为 0.5 至 2.5ng/mL。使用 2 个内部和 2 个外部 QC 样品测定所有化合物（喷他佐辛和 EDDP 除外，<20%）的日内和日间精密度均<15%，偏差在±15%以内（除甲基可卡因而外，<20%）。除 EDDP 外，所有化合物的提取效率均大于 70%。使用真实血液样本（n=10）评估基质效应，范围为 47%至 95%，但大多数分析物使用氘代类似物作为内标进行补偿。制备好的样品在自动进样器中可稳定 62 小时。该方法成功应用于真实样品（n=120），涉及海洛因、可卡因、曲马多和美沙酮的使用，以及能力验证计划。
