Simultaneous UHPLC-MS/MS method of estradiol metabolites to support the evaluation of Phase-2 metabolic activity of induced pluripotent stem cell derived hepatocytes

Simultaneous Quantification of Steroid Hormones Using hrLC-MS in Endocrine Tissues of Male Rats and Human Samples

“…The urine matrix does not impair the extraction of the non-sulfated steroids but the sulfated species suffer a recovery efficiency decay. To note, the studies measuring steroids in urine and tissues, as biological matrices report recovery efficiencies of over 100% in some of the cases [17][18][19]. An explanation for this phenomenon might be that the metabolites can be either free in solution or tethered to other molecules, such as membranal lipids during the extraction process.…”

“…The time required to perform the chromatographic separation is typically long in the literature; they report runtimes from over 10 min up to 45 min [3,5,[18][19][20][21][22]27]. Only the work of Quanson et al [16] and Indapurkar et al [17] described a methodology with a short runtime (4 to 5 min); however, they tested and applied the method solely in cell matrices: PCa and induced pluripotent stem cell lines, respectively. Indapurkar et al [17] developed a methodology specific for estradiol-related metabolites and Quanson et al [16] measured androgenic steroids using an ultra-performance convergence chromatography.…”

“…Indeed, a number of methods to detect and quantify steroid hormones have been reported during the last two decades. Many of the studies describe methodologies to detect steroids from several biological sources: cell cultures [3,16,17]; urine samples [18][19][20]; animal tissues [21][22][23]; human serum [24][25][26]; human hair [27] and waste water [28,29]. In general, steroid metabolomics methodologies focus on profiling a specific set of metabolites of interest in targeted tissues (or in circulation) rather than analyzing steroidogenesis status in a system of organs and related fluids.…”