Female and male serum reference intervals for challenging sex and precursor steroids by liquid chromatography tandem mass spectrometry.

•Four challenging steroids are measured in serum by a novel validated LC-MS/MS assay.•The ECLIA for estradiol measurement was in good agreement with the LC-MS/MS method.•The RIA for estrone measurement was in severe disagreement with the LC-MS/MS method.•Estrogens and dihydrotestosterone varied with menopause in women but not with age in men.•17-Hydroxypregnenolone levels depended on sex, menopause in women and age in men.