Current strategies for quantification of estrogen in clinical research

Denver, Nina and Khan, Shazia and Homer, Natalie Z.M. and MacLean, Margaret R. and Andrew, Ruth (2019) Current strategies for quantification of estrogen in clinical research. Journal of Steroid Biochemistry and Molecular Biology. ISSN 0960-0760

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    Abstract

    Estrogens and their bioactive metabolites play key roles in regulating diverse processes in health and disease. In particular, estrogen and estrogenic metabolites have shown both protective and non-protective effects on disease pathobiology, implicating the importance of this steroid pathway in disease diagnostics and monitoring. All estrogens circulate in a wide range of concentrations, which in some patient cohorts can be extremely low. However, elevated levels of E2 are also reported in disease. For example, in pulmonary arterial hypertension (PAH) levels are elevated in men with idiopathic PAH and in postmenopausal women with PAH. Conventional immunoassay techniques have been under scrutiny for some time with their selectivity, accuracy and precision coming into question. Analytical methodologies such as gas and liquid chromatography coupled to single and tandem mass spectrometric approaches (GC-MS, GC-MS/MS, LC-MS and LC-MS/MS) have been developed to quantify endogenous estrogens and in some cases their bioactive metabolites in biological fluids such as urine, serum, plasma and saliva. Liquid-liquid or solid-phase extraction approaches are favoured with derivatization remaining a necessity for lower volumes of sample. The limits of quantitation of individual assays vary but are commonly in the range of 0.5 – 5 pg/mL for estrone and estradiol, with limits of their bioactive metabolites being higher. This review provides an overview of current approaches for measurement of estrogens in biological matrices by MS, highlighting the advances in this field and the challenges remaining for routine use in the clinical and research environment.