Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti, 1, 00197, Rome, Italy; Dipartimento Chimica e Tecnologia del farmaco, "Sapienza" Università di Roma, Piazzale Aldo Moro 5, 00161, Rome, Italy.
Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti, 1, 00197, Rome, Italy; ISSUL - Institute of Sport Sciences, University of Lausanne, Synathlon - Quartier Centre, 1015, Lausanne, Switzerland.
J Pharm Biomed Anal. 2021 Feb 20;195:113849. doi: 10.1016/j.jpba.2020.113849. Epub 2020 Dec 28.
Analytical procedures to detect the misuse of selective androgen receptor modulators in human urine, targeting either the parent drugs and/or their main metabolites, were developed and validated. In detail, 19 target compounds belonging to 9 different chemical classes were considered: arylpropionamide (i.e., andarine (S4), ostarine (S22), S1, S6, S9 and S23), diarylhydantoin (i.e., GLPG0492), indole (i.e., LY2452473, GSK2881078), isoquinoline-carbonyle (i.e., PF-02620414), phenyl-oxadiazole (i.e., RAD140), pyrrolidinyl-benzonitrile (i.e., LGD4033), quinolinone (i.e., LGD2226, LGD3303), steroidal (i.e., Cl-4AS-1, MK0773 and TFM-4AS-1), and tropanol (i.e., AC-262536 and ACP105) derivatives. The metabolites of the target compounds considered were enzymatically synthesized by using human liver microsomes. Sample pre-treatment included enzymatic hydrolysis followed by liquid-liquid extraction at neutral pH. The instrumental analysis was performed by ultra-high-performance liquid chromatography coupled to either high- or low-resolution mass spectrometry. Validation was performed according to the ISO 17025 and the World Anti-Doping Agency guidelines. The analyses carried out on negative samples confirmed the method's selectivity, not showing any significant interferences at the retention times of the analytes of interest. Detection capability was determined in the range of 0.1-1.0 ng/mL for the screening procedure and 0.2-1.0 ng/mL for the confirmation procedure (except for GLPG0492 and GSK2881078). The recovery was greater than 80 % for all analytes, and the matrix effect was smaller than 35 %. The method also matched the criteria of the World Anti-Doping Agency in terms of repeatability of the relative retention times (CV% < 1.0) and of the relative abundances of the selected ion transitions (performed only in the case of triple quadrupole, CV% < 15), ensuring the correct identification of all the analytes considered. Urine samples containing andarine, ostarine, or LGD4033 were used to confirm the actual applicability of the selected analytical strategies. All target compounds (parent drugs and their main metabolites) were detected and correctly identified.
开发并验证了用于检测人尿中选择性雄激素受体调节剂（靶向母体药物和/或其主要代谢物）滥用的分析程序。详细地说，考虑了 19 种属于 9 种不同化学类别的目标化合物：芳基丙酰胺（即，那屈肝素（S4）、奥沙那林（S22）、S1、S6、S9 和 S23）、二芳基乙内酰脲（即，GLPG0492）、吲哚（即，LY2452473、GSK2881078）、异喹啉羰基（即，PF-02620414）、苯并恶二唑（即，RAD140）、吡咯烷基苯甲腈（即，LGD4033）、喹啉酮（即，LGD2226、LGD3303）、甾体（即，Cl-4AS-1、MK0773 和 TFM-4AS-1）和托烷（即，AC-262536 和 ACP105）衍生物。使用人肝微粒体酶促合成了目标化合物的代谢物。样品预处理包括在中性 pH 值下进行酶解，然后进行液-液萃取。仪器分析通过超高效液相色谱与高分辨率或低分辨率质谱联用进行。验证符合 ISO 17025 和世界反兴奋剂机构的指南。对阴性样本进行的分析证实了该方法的选择性，在感兴趣的分析物的保留时间处没有显示出任何显著的干扰。在筛查程序中，检测限为 0.1-1.0ng/mL，在确认程序中为 0.2-1.0ng/mL（GLPG0492 和 GSK2881078 除外）。所有分析物的回收率均大于 80%，基质效应小于 35%。该方法在相对保留时间的重复性（CV%<1.0）和所选离子跃迁的相对丰度（仅在三重四极杆的情况下进行，CV%<15）方面也符合世界反兴奋剂机构的标准，确保了所有考虑的分析物的正确识别。使用含有那屈肝素、奥沙那林或 LGD4033 的尿液样本验证了所选分析策略的实际适用性。所有目标化合物（母体药物及其主要代谢物）均被检测并正确识别。
