Application of a GLP protocol for the measurement of 17β-estradiol and testosterone by LC-MS/MS in the H295R steroidogenesis assay, Test No 456

Regulators have concerns about environmental chemicals, such as agrochemicals and their metabolites, and how to recognise their potential ability to disrupt the endocrine system1. These concerns led to the publication of a series of OECD in vitro test guidelines for the assessment of potential endocrine disrupting chemicals (EDCs). We have developed an extraction procedure and tandem LC-MS method1 to support one of these tests, TG4562, which investigates compound effects on the synthesis of the steroid hormones Testosterone (T) and 17β-estradiol (E2). The test uses the human H295R adreno-carcinoma cell line, as it encodes for all the key enzymes in the steroidogenesis pathway.

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