LC-MS/MS Analysis of Serum Estrogens for Clinical Research

The two major biologically active estrogens in non-pregnant humans are 17β-estradiol (E2) and estrone (E1). E2 is produced primarily in the ovaries and testes by the aromatization of testosterone, whereas most of E1 is derived from androstenedione. E2 can be metabolized to E1 and conversion of E1 to E2 is also possible, making the measurement of both compounds desirable.

The greatest challenge when analyzing E2 and E1 is the requirement to measure down to low concentration levels for certain clinical research applications. Currently, some immunoassay techniques lack analytical sensitivity and more commonly selectivity, while published LC-MS/MS methods use large sample volumes with complex sample extraction, often including derivatization.

The aim of this presentation is to demonstrate the performance of the ACQUITY UPLC™ I-Class and Xevo™ TQ-Absolute for the quantification of estrogens for clinical research.

During this presentation you will also learn about the importance of chromatographic separation of isotopic species, and choice of internal standard to compensate for matrix effects and how to measure them.

Presenter: Robert Wardle (Senior Scientist, Waters Corporation)

Robert Wardle is currently a Senior Scientist with the Clinical Business Unit at Waters Corporation in Wilmslow, UK, performing research and development of applications and products for Clinical Research. His primary area of research is vitamins and in particular vitamin D metabolites, steroid hormones and in-depth knowledge of offline and online automation.

Robert joined Waters in 2010, playing a key role in reagent Kit verification activities before moving on to developing LC-MS/MS methods for a range of applications including vitamin D metabolites, estrogens and a large panel of steroid hormones, where he also included the complete LC-MS/MS workflow of these application areas through offline automation and LIMS integration.