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

Background: 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.