Determination of endocrine disruptors and endogenic androgens and estrogens in rat serum by high-performance liquid chromatography–tandem mass spectrometry

•Simultaneous quantification of free and conjugated steroids and endocrine disruptors.•Limits of quantification between 0.1 and 0.7ng/mL in rat serum.•Rapid extraction performed in less than 10min.•Study provides interesting information about toxicological patterns of EDCs on rats. To simultaneously measure some targeted endocrine disruptors and several forms of sex hormones in rat serum, an accurate analytical procedure was developed. First, a comparison between a polymeric-based solid-phase extraction (SPE) and a micro-extraction by packed sorbent was performed to choose the optimal method to extract and concentrate the analytes: bisphenol A, atrazine, vinclozolin metabolite, testosterone, androstenedione, estrone, estradiol, estrone-sulfate and glucuronide and estradiol-sulfate and glucuronide. The analyses were then performed by high-performance liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) with electrospray ionisation in positive and negative modes. The protocol based on SPE was validated using the ICH/2005 guidelines. The validation demonstrated good performance in terms of linearity (R2>0.99), recovery (71–90%) and repeatability (relative standard deviation: 1–18%). The method was sensitive with LOQ comprised between 0.1 and 0.4ng/ml for androgens and between 0.098 and 10.2ng/ml for estrogens. The results obtained on the serum of rats exposed to the targeted endocrine disruptors showed the suitability of this analytical strategy.