Steroid profiling of modern and archaeological human bones using targeted liquid chromatography-tandem mass spectrometry (LC–MS/MS)

Abstract

A growing body of research explores the characterisation of steroids preserved in hard tissues (e.g., teeth and bones) to reconstruct the functioning of endocrine systems. Mineralized tissues have been shown to reflect circulating hormone levels during the last decade of an individual’s life and, importantly, these molecular records persist over archaeological timescales, albeit the mechanisms of preservation and degradation are poorly known. Here, we applied a targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to detect and quantify steroid molecules in modern and archaeological human bone, from a set of individuals with different biological sex, age-at-death, and bone health. We quantified nine steroids, including progestogens (pregnenolone, 17α-hydroxypregnenolone, progesterone, and 17α-hydroxyprogesterone); androgens (dehydroepiandrosterone (DHEA), androstenedione, and testosterone); and glucocorticoids (corticosterone, cortisol, and cortisone). The three most abundant steroid signals detected were DHEA, cortisol, and pregnenolone, but while modern bones yielded a broad steroid profile, archaeological bones showed a narrower profile, dominated by progestogens. This study confirms that human bones accumulate and store a broad range of steroid molecules and therefore can be a promising matrix for archaeo- and palaeo-metabolomics. However, the variability of our results highlights the importance of elucidating the accumulation patterns and diagenetic pathways of steroids in hard tissues before utilizing those signals for any interpretation of past physiological conditions.