Methods for the analysis of steroids have long been of interest due to the multiple uses for such methods in medical applications, sports monitoring, and environmental science. The analysis of steroids involves inherent analytical hurdles due to their low biological concentrations, poor ionization efficiencies, and frequent occurrence of isomerism. One analytical technique that has been recently applied to steroid analysis is ion mobility spectrometry (IMS). While previous work has focused on the use of metal adduction and multimer formation to enhance separation through IMS analysis coupled to mass spectrometry (MS), this work furthers this approach by coupling IMS-MS with liquid chromatography (LC). Three different LC methods with varying tradeoffs between chromatographic resolution and run time were developed, with one of these achieving a resolution above 1.5 for all steroid isomers. These results also indicate that the coupling of LC to IMS-MS can increase the overall resolution of steroid isomers relative to what can be achieved by either LC or IMS alone. Furthermore, the use of LC and IMS in concert can allow for a more rapid analysis of steroid isomers than can be achieved by LC-MS alone. Finally, the IMS dimension provided for measurements of ion-neutral collision cross sections (CCSs), which were found to be in good agreement with previously reported measurements. Thus, this approach provides three complementary quantitative parameters (retention time, CCS, and mass-to-charge ratio) that can contribute the identification of analytes. Overall, the work presented here demonstrates the potential of coupling LC, IMS, and MS for the analysis of isomeric steroid hormones.

由于类固醇的分析方法在医学应用，运动监测和环境科学中有多种用途，因此长期以来一直引起人们的兴趣。由于类固醇的低生物浓度，较差的电离效率和频繁发生的异构现象，因此类固醇的分析涉及固有的分析障碍。最近已应用于类固醇分析的一种分析技术是离子迁移谱（IMS）。虽然先前的工作集中在使用金属加成和多聚体形成以通过与质谱（MS）耦合的IMS分析增强分离的工作，但这项工作通过将IMS-MS与液相色谱（LC）耦合来进一步推广此方法。开发了三种在色谱分离度和运行时间之间权衡取舍的不同液相色谱方法，其中一种对所有类固醇异构体的分离度均高于1.5。这些结果还表明，相对于单独使用LC或IMS而言，LC与IMS-MS的偶联可以提高类固醇异构体的总体分离度。此外，与单独使用LC-MS相比，同时使用LC和IMS可以更快速地分析类固醇异构体。最后，IMS尺寸用于离子中性碰撞截面（CCS）的测量，发现该尺寸与以前报道的测量高度吻合。因此，该方法提供了三个互补的定量参数（保留时间，CCS和质荷比），这些参数可以有助于分析物的鉴定。总体而言，此处介绍的工作证明了LC，IMS，