Paleohormones: A new biomolecular perspective on archaeological bone

<h2>Abstract</h2> There has been limited analysis of steroid hormones in archaeological skeletal material. What has been done has used enzyme-linked immunosorbent assays (ELISA), which should be critically viewed due to its known cross-reactivity leading to false positives and/or inaccurate results. Here, we (1) propose the first quantitative analysis of steroids in human bone, (2) verify the preservation of steroids in archaeological human bone, (3) are the first to apply liquid chromatography-tandem mass spectrometry (LC-MS/MS) to the analysis of paleohormones. LC-MS/MS provides high sensitivity and specificity, allowing the accurate quantification of closely related compounds. We analyzed archaeological femora and ribs from 13 individuals (1350–1829 CE) and quantified endogenous testosterone and progesterone, while other steroids (cortisol, cortisone, corticosterone, 11-dehydrocorticosterone, dihydrotestosterone and β-estradiol) could not be detected. Testosterone values in ribs significantly differ between males and females and may be used as a method of sex determination in the future.