Steroid profiling in nails using liquid chromatography-tandem mass spectrometry

•An LC-MS/MS method was developed for the quantification of 12 steroid hormones in nails.•3 steroid hormones were detected in nails for the first time.•Six steroid hormones were detected and quantified in human fingernails of mothers and their infants.•No significant differences could be found between right and left hand steroid levels. The retrospective analysis of endogenous steroid hormones in nails can be used to elucidate endocrine diseases and thus help with their diagnosis and treatment. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) based method was developed for the simultaneous identification and quantification of 12 steroid hormones (aldosterone, cortisone, cortisol, corticosterone, 11-deoxycortisol, androstenedione, 11-deoxycorticosterone, testosterone, dehydroepiandrosterone (DHEA), 17α-hydroxyprogesterone (17-OHP), dihydrotestosterone (DHT) and progesterone) in human fingernails. Steroid hormones were extracted from 0.5 mg to 10 mg pulverized nail clippings by methanolic extraction, followed by a liquid-liquid extraction. The analysis was conducted with LC-MS/MS in electrospray ionization positive mode. The method was validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, matrix effect, recovery and robustness. It was successfully applied for steroid profiling in nails of mothers and their infants where cortisol, cortisone, testosterone, progesterone, androstenedione and 11-deoxycorticosterone could be detected. Furthermore, it could be shown that there is no significant difference in concentrations between left and right hand for cortisol, cortisone and progesterone. A positive linear correlation between cortisol and cortisone in nails was found. In conclusion, it could be shown that nails are a suitable matrix for the retrospective monitoring of cumulative steroid hormone levels.