Urinary Sex Steroids and Anthropometric Markers of Puberty - A Novel Approach to Characterising Within-Person Changes of Puberty Hormones

Abstract

Background/AimsThe longitudinal relationships of within-individual hormone and anthropometric changes during puberty have not ever been fully described. The objectives of this study were to demonstrate that 3 monthly urine collection was feasible in young adolescents and to utilise liquid chromatography-tandem mass spectrometry assay methods for serum and urine testosterone (T), estradiol (E2) and luteinizing hormone (LH) in adolescents by relating temporal changes in urine and serum hormones over 12 months to standard measures of pubertal development.MethodsA community sample of 104 adolescents (57 female) was studied over 12 months with annual anthropometric assessment, blood sampling and self-rated Tanner staging and urine collected every 3 months. Serum and urine sex steroids (T, E2) were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and LH by immunoassay.ResultsA high proportion (92%) of scheduled samples were obtained with low attrition rate of 6.7% over the 12 months. Urine hormone measurements correlated cross-sectionally and longitudinally with age, anthropometry and Tanner stage.ConclusionWe have developed a feasible and valid sampling methodology and measurements for puberty hormones in urine, which allows a sampling frequency by which individual pubertal progression in adolescents can be described in depth.

Foot Length Growth as a Novel Marker of Early Puberty

“…Following this, tandem-mass spectrometry analysis was performed using an API-5000 triple-quadrupole mass spectrometer (Applied Biosystem/MDS SCIEX, Ontario, Canada). 6,14 This method can quantify low levels of circulating E 2 and T in 99% of serum specimens from adolescents. 6…”

“…Early morning serum E 2 (collected in follicular phase for post-menarcheal participants) and T were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using previously described methods. 6,14 Briefly, organic solvent extracts of serum were separated by liquid chromatography using a Shimadzu Nexera UHPLC system (Shimadzu Scientific Instruments, Columbia, MD). Following this, tandem-mass spectrometry analysis was performed using an API-5000 triple-quadrupole mass spectrometer (Applied Biosystem/MDS SCIEX, Ontario, Canada).…”

“…Peak height velocity (PHV) requires serial calibrated measurements with sufficient frequency to determine a clear peak. Sex steroid measurement directly samples the hormonal milieu that drives the observed physical changes of puberty, but is limited due to acceptability of repeat biological sampling with adequate frequency 6 and the assay cost making this approach less useful for population-based assessment of puberty. Thus, a marker that is simple and easily measured, cost-effective, correlates with established markers of puberty (especially sex steroids), and which additionally may indicate the early stages of puberty is likely to be of significant research and clinical utility.…”

Methods for determining pubertal status in research studies: literature review and opinions of experts and adolescents

“…In a study examining the correlation of three-monthly urinary oestradiol, testosterone and LH with self-reported SMS-based Tanner staging, the levels of all three hormones were correlated with the staging at baseline and 12 months later. 43 In another study, staging based on physical examination reflected testosterone and DHEA levels in both sexes, but was only modestly related to oestradiol in girls. 44 In boys, 24-hour testosterone levels in later puberty correlate reasonably well with testicular volume.…”

“…45 However, assays of urinary sex hormones both in boys and girls can be difficult to interpret due to within-person variability, and longitudinal sampling would be required to determine the hormonal changes associated with progression through pubertal stages. 43,46 It has been suggested that both inhibin A and B may have a potential as markers of pubertal development in boys and girls. 46,47 However, progressive changes in their levels are not consistent enough to enable pubertal staging.…”