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University of Sheffield and University College London researchers have made the first successful detection of estrogen, progesterone, and testosterone in bones, teeth and dental calculus, opening a way to identify pregnancy in the archaeological record.

Archaeologists face major gaps in reconstructing reproductive histories. Direct identification of pregnancy relies on rare in utero fetal remains, and indirect pelvic markers lack reliability. Hard tissues survive far more often than soft tissues or hair, requiring new biochemical strategies to uncover pregnancies of the past.

In the study, "First successful detection of oestrogen, progesterone and testosterone in archaeological human hard tissues, and their use as potential biomarkers of pregnancy," in Journal of Archaeological Science, researchers developed a novel ELISA-based extraction and analysis protocol to test whether sex steroids can be recovered from bone, tooth structures, and dental calculus and assessed as pregnancy biomarkers.

Ten individuals of known sex (via DNA or osteologically), dating from the 1st鈥�19th centuries, formed the cohort, including three males and seven females sampled across multiple British cemeteries.

Of the females, two were found with in utero fetal remains, two were buried with fetal remains (mother-child burials), one historically inferred mother (confirmed via DNA), and two with unknown childbirth histories.

Methodology centered on methanol extraction of powdered bone, dentin, enamel, roots from second and third permanent molars, and dental calculus, yielding 74 prepared samples. Assays targeted 17尾-estradiol, progesterone, and testosterone.

Hormones were discovered to be measurable in bone, dentin, enamel, and root. Progesterone appeared in 24.3% of samples, testosterone in 46.6%, and estrogen in 9.5%.

Dental calculus yielded progesterone and testosterone only. Bone provided the most frequent progesterone positives. One pregnant female showed elevated progesterone in bone and tooth structures, including in a vertebral fragment, while another had bone progesterone but no tooth progesterone.

Testosterone was undetectable from in utero pregnancies across tested tissues. Females buried with associated fetal remains showed elevated progesterone in calculus. Enamel and dentin sometimes carried similar steroid concentrations, challenging previous assumptions about enamel's lower organic fraction dictating a lower hormone signal.

Authors conclude that an ELISA workflow can recover sex steroids from archaeological hard tissues and that steroid profiles hold promise for identifying pregnancy at death and for reconstructing aspects of female life histories.

Broader application could expand visibility of pregnant individuals in the archaeological record and support new questions about hormone deposition, tissue physiology, and reproductive timelines in past populations.

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