Laboratory of Toxicology, KULeuven, Campus Gasthuisberg O&N 2, PO Box 922, Herestraat 49, 3000 Leuven, Belgium.
Forensic Sci Int. 2012 Feb 10;215(1-3):136-45. doi: 10.1016/j.forsciint.2011.01.047. Epub 2011 Feb 26.
Narcotic analgesics are widely (ab) used and sometimes only occur in low concentrations in biological samples. Therefore, a highly sensitive liquid chromatography tandem mass spectrometry method was developed for simultaneous analysis of 9 narcotic analgesics and metabolites (buprenorphine, O-desmethyltramadol, fentanyl, norbuprenorphine, norfentanyl, pethidine, piritramide, tilidine and tramadol) in urine and whole blood. Sample preparation was performed on a mixed-mode cation exchange solid phase extraction cartridge with an additional alkaline wash step to decrease matrix effects and thus increase sensitivity. Ionization with electrospray ionization was found to be more efficient than atmospheric pressure chemical ionization. The use of a mobile phase of high pH resulted in higher electrospray ionization signals than the conventional low pH mobile phases. In the final method, gradient elution with 10mM ammonium bicarbonate (pH 9) and methanol was performed on a small particle column (Acquity C18, 1.7 μm, 2.1 mm × 50 mm). Selectivity, matrix effects, recovery, linearity, sensitivity, precision, accuracy and stability were validated in urine and whole blood. All parameters were successfully evaluated and the method showed very high sensitivity, which was the major aim of this study. The applicability of the method was demonstrated by analysis of several forensic cases involving narcotic analgesics.
麻醉性镇痛药被广泛（滥用）使用，有时仅以低浓度存在于生物样本中。因此，开发了一种高灵敏度的液相色谱串联质谱法，用于同时分析尿液和全血中的 9 种麻醉性镇痛药及其代谢物（丁丙诺啡、O-去甲曲马多、芬太尼、去甲丁丙诺啡、去甲芬太尼、哌替啶、喷他佐辛、替利定和曲马多）。样品制备采用混合模式阳离子交换固相萃取小柱进行，增加碱性洗涤步骤以降低基质效应，从而提高灵敏度。电喷雾电离比大气压化学电离更有效。使用高 pH 值的流动相可产生比传统低 pH 值流动相更高的电喷雾电离信号。在最终方法中，采用 10mM 碳酸氢铵（pH 9）和甲醇在小颗粒柱（Acquity C18，1.7μm，2.1mm×50mm）上进行梯度洗脱。在尿液和全血中对选择性、基质效应、回收率、线性、灵敏度、精密度、准确度和稳定性进行了验证。所有参数均成功评估，该方法具有很高的灵敏度，这是本研究的主要目标。通过对涉及麻醉性镇痛药的几个法医案例的分析，证明了该方法的适用性。
