Verri Patrizia, Rustichelli Cecilia, Palazzoli Federica, Vandelli Daniele, Marchesi Filippo, Ferrari Anna, Licata Manuela
Department of Diagnostic Medicine, Clinical and Public Health, Forensic Toxicology Laboratory, University of Modena and Reggio Emilia, via del Pozzo 71, 41124 Modena, Italy.
Department of Life Sciences, University of Modena and Reggio Emilia, via G. Campi 183, 41125 Modena, Italy.
J Pharm Biomed Anal. 2015 Jan;102:450-8. doi: 10.1016/j.jpba.2014.10.002. Epub 2014 Oct 12.
Hair analysis, as complementary matrix, has expanded across the spectrum of toxicological investigations for misuse drug monitoring. Hair has become an important matrix for drug analysis, owing to the possibility to detect target analytes for long time periods, depending on hair length. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed for the quantitation of tramadol, a widely used centrally acting analgesic, and its main metabolites in hair (ODMT, NDMT, NOT). Hair samples were decontaminated and incubated overnight in diluted hydrochloric acid; the extracts were purified by mixed-mode solid phase cartridges and analyzed by LC-MS/MS in positive ionization mode monitoring two transitions per analyte. The procedure was fully validated in terms of linearity, limit of detection and lower limit of quantitation (LLOQ), accuracy, precision, recovery, matrix effect and selectivity. The linear regression analysis was calibrated by deuterated internal standards; for all analytes, responses were linear over the range 0.04-40.00 ng/mg hair, with R(2) values of at least 0.995. The method offered satisfactory precision (RSD < 10%), accuracy (90-110%) and recovery (> 90%) values. The found LLOQ values for tramadol and metabolites were in the range 0.010-0.030 ng/mg hair. The proposed procedure was successfully applied to quantify tramadol and metabolites in real hair samples submitted to our laboratory: three cases of tramadol assumption within the therapeutic dosage (3 × 2 segments) and one case of tramadol abuse in a binge pattern (8 segments). The ranges found for TRAM, ODMT, NDMT and NOT were markedly higher in the abuse case (63.42-107.30, 3.76-6.26, 24.88-45.66, 0.22-1.18 ng/mg hair, respectively) compared to the other case reports (3.29-20.12, 0.28-1.87, 0.45-4.32, 0.07-0.80 ng/mg, respectively); also the values of NMDT/ODMT ratio differed significantly. According to the obtained data, we hypothesized that the binge pattern may influence the metabolites' to parent drug concentration ratios; therefore this parameter could represent a target assessment tool to monitor abuse cases.
毛发分析作为一种补充基质，已在药物滥用监测的毒理学调查领域广泛应用。由于根据毛发长度能够长时间检测目标分析物，毛发已成为药物分析的重要基质。已开发出一种液相色谱 - 串联质谱法（LC - MS/MS）用于定量检测毛发中的曲马多（一种广泛使用的中枢性镇痛药）及其主要代谢物（ODMT、NDMT、NOT）。毛发样品经过去污处理后，在稀盐酸中孵育过夜；提取物通过混合模式固相萃取柱进行纯化，并采用LC - MS/MS在正离子模式下进行分析，每个分析物监测两个跃迁。该方法在线性、检测限和定量下限（LLOQ）、准确度、精密度、回收率、基质效应和选择性方面均得到了充分验证。线性回归分析采用氘代内标进行校准；对于所有分析物，在0.04 - 40.00 ng/mg毛发的范围内响应呈线性，R(2)值至少为0.995。该方法具有令人满意的精密度（RSD < 10%）、准确度（90 - 110%）和回收率（> 90%）。所测得的曲马多及其代谢物的LLOQ值在0.010 - 0.030 ng/mg毛发范围内。所提出的方法成功应用于对提交至我们实验室的实际毛发样品中的曲马多及其代谢物进行定量分析：三例治疗剂量内的曲马多摄入情况（3×2段毛发）以及一例暴饮暴食模式下的曲马多滥用情况（8段毛发）。与其他病例报告相比（分别为3.29 - 20.12、0.28 - 1.87、0.45 - 4.32、0.07 - 0.80 ng/mg），滥用病例中TRAM、ODMT、NDMT和NOT的含量范围明显更高（分别为63.42 - 107.30、3.76 - 6.26、24.88 - 45.66、0.22 - 1.18 ng/mg毛发）；NMDT/ODMT比值也存在显著差异。根据所获得的数据，我们推测暴饮暴食模式可能会影响代谢物与母体药物的浓度比；因此，该参数可能是监测滥用病例的一种目标评估工具。
