Determination of 17OHPreg and DHEAS by LC-MS/MS: Impact of Age, Sex, Pubertal Stage, and BMI on the Δ5 Steroid Pathway

J Clin Endocrinol Metab. 2017 Jan 1;102(1):232-241. doi: 10.1210/jc.2016-2849.

Abstract

Background: Dehydroepiandrosterone sulfate (DHEAS) and 17-hydroxypregnenolone (17OHPreg) are important for understanding the Δ5 pathway (e.g., in adrenarche and obesity). Although mass spectrometry has become the state-of-the-art method for quantifying steroids, there are few comprehensive age-, sex-, and pubertal stage-specific reference ranges for children.

Aims: To develop a sensitive and reliable ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for simultaneous quantification of DHEAS and 17OHPreg and to establish entire age-, sex- and pubertal stage-specific reference ranges in children.

Methods: A total of 684 children, 453 (243 female, 210 male) with normal body mass index (BMI; <90th) and 231 (132 female, 99 male) obese subjects (>97th), were categorized into 11 age groups, and age- and Tanner stage (PH)-specific reference ranges were determined.

Results: The limit of detection was 0.05 nmol/L for 17OHPreg and 0.5 nmol/L for DHEAS. Levels of both steroids declined after the neonatal period. Comparisons with RIA assays (Siemens, Munich, Germany) (DHEAS) and an in-house kit (17OHPreg) revealed 0.95 and 0.93, respectively, as coefficients of determination. Although DHEAS-generally higher in boys-increased continuously starting at 3 to 6 years, 17OHPreg remained largely constant. In obese patients, both were significantly elevated, also in part after alignment to Tanner stages (PH).

Conclusions: UPLC-MS/MS is sensitive and reliable for quantifying DHEAS and 17OHPreg. Our data support differential maturation of CYP17 during adrenarche with successively increasing 17,20-lyase activity but largely constant 17α-hydroxylation activity. Endocrine interpretation of 17OHPreg and DHEAS must consider differential patterns for age, sex, pubertal stage, and BMI.

Publication types

  • Comparative Study

MeSH terms

  • 17-alpha-Hydroxypregnenolone / analysis*
  • Adolescent
  • Age Factors
  • Biomarkers / analysis
  • Body Mass Index
  • Case-Control Studies
  • Child
  • Child, Preschool
  • Chromatography, Liquid / methods*
  • Dehydroepiandrosterone Sulfate / analysis*
  • Female
  • Follow-Up Studies
  • Humans
  • Infant
  • Infant, Newborn
  • Male
  • Obesity / physiopathology*
  • Prognosis
  • Puberty / metabolism*
  • Sex Factors
  • Steroid 17-alpha-Hydroxylase / metabolism
  • Steroids / metabolism*
  • Tandem Mass Spectrometry / methods*

Substances

  • Biomarkers
  • Steroids
  • 17-alpha-Hydroxypregnenolone
  • Dehydroepiandrosterone Sulfate
  • Steroid 17-alpha-Hydroxylase