Institute of Biochemistry-Center for Preventive Doping Research, German Sport University Cologne, Cologne, Germany.
Anal Bioanal Chem. 2011 Aug;401(2):543-51. doi: 10.1007/s00216-010-4455-4. Epub 2010 Nov 30.
Benfluorex [1-(m-trifluoromethylphenyl)-2-(β-benzoyloxyethyl)aminopropane] has been widely used for the treatment of atherogenic metabolic disorders and impaired carbohydrate metabolism (particularly in obese type-II diabetic patients) as well as an anorectic drug. Due to its potentially performance-enhancing properties, benfluorex has been added to the list of prohibited compounds and methods of doping by the World Anti-Doping Agency (WADA) in 2010, necessitating the implementation of the drug as well as its major metabolites into routine doping control procedures. In the present study, human urinary metabolites of benfluorex were characterized by gas chromatography-electron ionization-mass spectrometry (GC-EI-MS) as well as liquid chromatography-electrospray ionization-high resolution/high accuracy tandem mass spectrometry (LC-ESI-MS/MS). Commonly employed sports drug testing approaches consisting of liquid-liquid extraction followed by GC-MS or urine dilution and immediate LC-MS/MS analysis were expanded and validated with regard to specificity, recovery (48-54%, GC-MS only), intra- and interday precision (<25%), limits of detection (5-8 ng/mL for LC-MS/MS and 80 ng/mL for GC-MS), and ion suppression (for LC-ESI-MS/MS only) to allow the detection of benfluorex metabolites 1-(m-trifluoromethylphenyl)-2-(2-hydroxyethyl)aminopropane (M1), 1-(m-trifluoromethylphenyl)-2-(2-carboxymethyl)aminopropane (M2), and 1-(m-trifluoromethylphenyl)-2-aminopropane (M3) as well as the glucuronic acid conjugate of M1.
苯氟雷司[1-(间三氟甲基苯基)-2-(β-苯甲酰氧基乙基)氨基丙烷]被广泛用于治疗动脉粥样硬化代谢紊乱和碳水化合物代谢受损（尤其是肥胖型 2 型糖尿病患者）以及作为食欲抑制剂。由于其具有潜在的增强性能，苯氟雷司于 2010 年被世界反兴奋剂机构（WADA）列入禁用化合物和方法名单，需要将该药物及其主要代谢物纳入常规兴奋剂控制程序。在本研究中，采用气相色谱-电子电离-质谱（GC-EI-MS）和液相色谱-电喷雾电离-高分辨/高精度串联质谱（LC-ESI-MS/MS）对人尿中的苯氟雷司代谢物进行了表征。常用的运动药物检测方法包括液-液萃取后进行 GC-MS 或尿液稀释和即时 LC-MS/MS 分析，本研究对这些方法进行了扩展和验证，以确保其特异性、回收率（GC-MS 仅为 48-54%）、日内和日间精密度（<25%）、检测限（LC-MS/MS 为 5-8ng/mL，GC-MS 为 80ng/mL）和离子抑制（仅适用于 LC-ESI-MS/MS），以允许检测苯氟雷司代谢物 1-(间三氟甲基苯基)-2-(2-羟乙基)氨基丙烷（M1）、1-(间三氟甲基苯基)-2-(2-羧甲基)氨基丙烷（M2）和 1-(间三氟甲基苯基)-2-氨基丙烷（M3）以及 M1 的葡萄糖醛酸缀合物。
