Assay validation of hair androgens across the menstrual cycle

Saliva is a common noninvasive biofluid for measuring stress and sex hormones, yet one pressing limitation is that salivary hormones fluctuate momentarily, daily, and (for girls) across the menstrual cycle. Hair steroid assays are thought to provide a cumulative index which collapses across hormonal variability, potentially eliminating the confound of daily and menstrual cyclicity and thereby reflecting individual differences in average hormone levels. Here we seek to validate a hair bioassay methodology and test whether hair androgens accurately measure long-term, stable androgen levels in emerging adult women across two menstrual cycles.

Hair samples were collected at the end of each menstrual cycle for two cycles, and saliva samples were collected in the morning once per week across two menstrual cycles (N = 11 women). Hair samples were segmented by 1 cm for the first 4 cm to reflect the hormone levels of the past four serial months. Hair samples were assayed using commercially-available enzyme-immuno-assays for testosterone and DHEA.

Hair androgen concentrations were significantly correlated with averaged saliva hormone levels (DHEA: r = .75, p < .05; Testosterone: r = .67, p < .05). With respect to hair hormone stability, there were significant correlations for almost all the pairs of two 1 cm hair segments collected in two months that corresponded to the same time period. Hair androgens in one segment were significantly correlated with those in next segment. Regarding salivary androgen stability, the intra-class correlation across the weekly saliva samples indicated that for DHEA 59% of the total variance was within person and 41% was between person; and for testosterone 91% of the total variance was between person, and only 9% within person.

Results suggest that a one-time measure of hair provides a valid and reliable estimate of average steroid levels across two months. Moreover, whereas saliva measures of androgen levels capture week-to-week fluctuations in steroids, hair samples provide information on individual differences in average exposure to steroids, across long periods of time, such as months. Results are encouraging that hair DHEA and testosterone reflects the cumulative hormonal concentration and can be used as a stable hormonal index. Results also indicate that it is feasible to collect the first 3-4 centimeters of hair for studies of stable hormone levels.