Steroids in Reproduction: LC-MS-MS and GC (-MS) Based Steroidomics

以最高的可靠性测定类固醇硫酸盐是实现研究单位硫化类固醇在生殖中的先决条件。CS、PREGS、DHEAS和E1S将是要求的主要分析物。用LC-MS-MS对化合物进行定性和定量测定，为分析完整的共轭甾体提供了一线的分析方法。由于除DHEAS外，尚不存在基于LC-MS-MS的适用程序，我们的首要目标将是利用样品制备和质谱分析方面的最新技术发展和改进，开发一种用于上述类固醇结合物的多靶点代谢图谱的分析方法。我们实验室精良的设备和目前可用的最好的新型LC-MS-MS系统(TSQ Qantum Ultra，ThermoFisher)将允许开发基于高选择性TurboFlow-FAIMS-MS的分析方法。与使用稳定同位素标记的分析物类似物作为内部标准一样，这种分析方法将允许在代谢水平上最大限度地改善对生殖机制的阐明。在第二阶段，这一新开发的分析方法将应用于合作主要研究人员的个人研究提案中提到的生物样本的分析。

Determination of steroid sulfates with highest reliability is a prerequisite for the realization of theResearch Unit Sulfated Steroids in Reproduction. CS, PREGS, DHEAS and E1S will be the mainanalytes requested. It is planned to carry out the qualitative and quantitative determination of thesecompounds by liquid chromatography-tandem mass spectrometry (LC-MS-MS), which currentlypresents the analytical first line approach for the analysis of the intact conjugated steroids. Since –with the exception of DHEAS – suitable procedures based on LC-MS-MS do not exist, our first aimwill consist in developing an analytical method for multi-targeted metabolic profiling of the abovementioned steroid conjugates using latest technical developments and improvements in samplepreparation and mass spectrometric analysis. Excellent equipment of our laboratory with one of thebest currently available new LC-MS-MS systems (TSQ Qantum Ultra, ThermoFisher) will permitdevelopment of a highly selective TurboFlow-FAIMS-MS-MS based analytical procedure. Togetherwith the use of stable isotope labeled analogs of the analytes as internal standards, this analyticalapproach will allow for a maximally improved elucidation of reproductive mechanisms at themetabolic level. In a second stage, this newly developed analytical method will be applied to theanalysis of biological samples mentioned in the individual research proposals of the collaboratingprincipal investigators.