LC‐MS/MS‐based method for long‐term steroid profiling in human scalp hair

SummaryBackground Liquid chromatography–tandem mass spectrometry (LC-MS/MS) is the method of choice for quantification of steroids. Human scalp hair provides the possibility to measure long-term retrospective steroid concentrations, which is especially useful for steroids with large time-dependent fluctuations in concentration, such as the glucocorticoid cortisol. Aim We set up and validated a LC-MS/MS-based method for long-term steroid profiling, quantifying cortisol, cortisone, testosterone, androstenedione, dehydroepiandrosterone sulphate (DHEAS) and 17-α-hydroxyprogesterone (17OHP). Method Hair locks were cut from the posterior vertex of healthy male and female volunteers and washed in isopropanol. Steroids were extracted using methanol, and extract was cleaned up by solid-phase extraction and measured on a Waters XEVO-TQ-S LC-MS/MS. Lower limit of quantification, precision, matrix interference and intra-individual variation were determined. Results The functional sensitivity of our steroid analysis was <1·3, <9·3, 2·3, <1·3, <15·9, 1·87 pg/mg hair for cortisol, cortisone, testosterone, androstenedione, dehydroepiandrosterone sulphate (DHEAS), and 17OHP, respectively. Measured over a 9-month period, the inter-run CVs were below 16% for all steroids. Intra-individual coefficients of variation were below 15% for all steroids, except 17OHP (19·7%). Conclusion The authors present a LC-MS/MS-based method for long-term steroid profiling in human scalp hair, potentially providing novel insights by a multitude of clinical and research applications in the field of endocrinology.