Development and validation of a liquid chromatography coupled to mass spectrometer (LC-MS) method for the simultaneous quantification of estrone-3-sulfate, progesterone, estrone and estradiol in serum of mares and American bisons

Steroid concentrations in serum are fluctuating during pregnancy of many mammal species. The current knowledge about endocrinology gestation is mainly based on immunoassays. However, the lack specificity these assays hampers reliability results. In present work, we developed and validated a methodology associating liquid chromatography (LC) mass spectrometry (MS) to simultaneously quantify, with high accuracy, estrone-3-sulfate (E3S), progesterone (PRO), estrone (E1) estradiol (E2) two different sample preparation procedure simple protein precipitation derivatization dansyl chloride. After chromatographical separation, compounds were analyzed triple-quadrupole spectrometer operating multiple reaction monitoring. Mare American bison samples method results compared measured commercial radioimmunoassay (RIA), enzyme linked immunosorbent assay (ELISA) chemiluminescent microparticle immunoassay (CMIA). Following criterions: relative standard deviation <15% bias <15%, lower limits quantification 0.5 ng/mL 0.1 (PRO) 2 pg/mL (E1 E2) achieved. Most comparison between immunoassays LC-MS showed poor correlation proportional differences. Our able quantify several steroid hormones specificity, accuracy sensitivity constitutes useful performant tool for veterinary clinicians should thus be used update refine mammals.