Plachká Kateřina, Pezzatti Julian, Musenga Alessandro, Nicoli Raul, Kuuranne Tiia, Rudaz Serge, Nováková Lucie, Guillarme Davy
Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Heyrovského 1203, 500 05, Hradec Králové, Czech Republic.
School of Pharmaceutical Sciences, University of Geneva, CMU-Rue Michel Servet 1, 1211, Geneva 4, Switzerland; Institute of Pharmaceutical Sciences of Western Switzerland (ISPSO), University of Geneva, CMU-Rue Michel Servet 1, 1211, Geneva 4, Switzerland.
Anal Chim Acta. 2021 Apr 1;1152:338257. doi: 10.1016/j.aca.2021.338257. Epub 2021 Jan 31.
In this series of two papers, 192 doping agents belonging to the classes of stimulants, narcotics, cannabinoids, diuretics, β2-agonists, β-blockers, anabolic agents, and hormone and metabolic modulators were investigated, with the aim to assess the benefits and limitations of ion mobility spectrometry (IMS) in combination with ultra-high performance liquid chromatography (UHPLC) and high resolution mass spectrometry (HRMS) in anti-doping analysis. In this first part, a generic UHPLC-IM-HRMS method was successfully developed to analyze these 192 doping agents in standard solutions and urine samples, and an exhaustive database including retention times, CCS values, and m/z ratios was constructed. Urine samples were analyzed using either a simple "dilute and shoot" procedure or a supported liquid-liquid extraction (SLE) procedure, depending on the physicochemical properties of the compounds and sensitivity criteria established by the World Anti-Doping Agency (WADA) as the minimum required performance levels (MRPL). Then, the precision of the generic UHPLC-IM-HRMS method was assessed as intraday, interday as well as interweek variation of UHPLC retention times and CCS values, for which RSD the values were always lower than 2% in urine samples. The possibility to filter MS data using IMS dimension was also investigated, and in average, the application of IMS filtration provided low energy MS spectra with 86% less interfering peaks in both standard and urine samples. Therefore, the filtered MS spectra allowed for an easier interpretation and a lower risk of false positive result interpretations. Finally, IMS also offers additional selectivity to the UHPLC-HRMS enabling to separate isobaric and isomeric substances. Among the selected set of 192 doping agents, there were 30 pairs of isobaric or isomeric compounds, and only two pairs could not be resolved under the developed conditions. This illustrates the potential of adding ion mobility to UHPLC-HRMS in anti-doping analyses.
在这两篇系列论文中，对属于兴奋剂、麻醉品、大麻素、利尿剂、β2激动剂、β阻滞剂、同化剂以及激素和代谢调节剂类别的192种掺杂剂进行了研究，目的是评估离子淌度光谱法（IMS）与超高效液相色谱法（UHPLC）和高分辨率质谱法（HRMS）相结合在反兴奋剂分析中的优势和局限性。在第一部分中，成功开发了一种通用的UHPLC-IM-HRMS方法，用于分析标准溶液和尿液样本中的这192种掺杂剂，并构建了一个详尽的数据库，其中包括保留时间、CCS值和质荷比。根据化合物的物理化学性质以及世界反兴奋剂机构（WADA）确定的作为最低要求性能水平（MRPL）的灵敏度标准，使用简单的“稀释进样”程序或支持液-液萃取（SLE）程序对尿液样本进行分析。然后，评估通用UHPLC-IM-HRMS方法的精密度，以UHPLC保留时间和CCS值的日内、日间以及周间变化来衡量，尿液样本中的相对标准偏差（RSD）值始终低于2%。还研究了使用IMS维度过滤质谱数据的可能性，平均而言，IMS过滤的应用在标准和尿液样本中提供了干扰峰减少86%的低能量质谱图。因此，过滤后的质谱图更易于解释，且出现假阳性结果解释的风险更低。最后，IMS还为UHPLC-HRMS提供了额外的选择性，能够分离等压和同分异构物质。在选定的192种掺杂剂中，有30对等压或同分异构化合物，在已开发的条件下只有两对无法分离。这说明了在反兴奋剂分析中，将离子淌度添加到UHPLC-HRMS中的潜力。
