Determination of 17OHPreg and DHEAS by LC-MS/MS: Impact of Age, Sex, Pubertal Stage, and BMI on the [Delta]5 Steroid Pathway

• Published in: The journal of clinical endocrinology and metabolism 2017-01, Vol.102 (1), p.232
• Main Authors: Kulle, Alexandra E, Reinehr, Thomas, Simic-Schleicher, Gunter, Hornig, Nadine C, Paul-Martin, Holterhus
• Format: Article
• Language: English
• Subjects: Age; Body mass; Body mass index; Children; Dehydroepiandrosterone; Dehydroepiandrosterone sulfate; Hydroxylation; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Neonates; Obesity; Scientific imaging; Sex; Steroid hormones; Steroids; Sulfates
• ISSN: 0021-972X; 1945-7197
• DOI: 10.1210/jc.2016-2849

Précis: We developed a UPLC-MS/MS method for measurement of 17OHPreg and DHEAS. Reference ranges based on age, sex, and pubertal stage were determined. The impact of BMI on the Δ5 pathway was studied. Abstract 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. 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. A total of 684 children, 453 (243 female, 210 male) with normal body mass index (BMI; 97th), were categorized into 11 age groups, and age- and Tanner stage (PH)-specific reference ranges were determined. 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). 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.