Implementation of a liquid chromatography tandem mass spectrometry assay for eight adrenal C-21 steroids and pediatric reference data

Horm Res Paediatr. 2013;79(1):22-31. doi: 10.1159/000346406. Epub 2013 Jan 17.

Abstract

Background: Sensitive and accurate determination of steroids is essential for diagnosing congenital and acquired adrenal diseases. Since plasma concentrations change during childhood, age-specific reference ranges are the prerequisite for clinical interpretation. The objectives of this study were to develop a sensitive and reliable method for simultaneous detection and quantification of progesterone, 17-hydroxyprogesterone, deoxycorticosterone (DOC), 11-deoxycortisol, 21-deoxycortisol, corticosterone, cortisol (F) and cortisone (E) by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) and to establish age- and sex-specific reference ranges from birth to adulthood.

Methods: All eight steroids were measured simultaneously in 0.1 ml plasma by UPLC-MS/MS. Samples of 905 children were measured and grouped in five age groups.

Results: The assay was linear up to 70 ng/ml (700 ng/ml for F; r(2) > 0.992). The limit of detection ranged between 0.01 ng/ml for DOC and 0.07 ng/ml for E. Correlations with radioimmunoassays yielded a coefficient of determination between 0.82 and 0.99. Reference data are reported as a function of age and sex.

Conclusions: The UPLC-MS/MS method presented here for the simultaneous detection of eight C-21 adrenal hormones together with the detailed reference ranges for children provides a valuable methodology for assessing adrenal steroids in clinical routine and research.

Publication types

  • Evaluation Study

MeSH terms

  • 17-alpha-Hydroxyprogesterone / blood
  • Adolescent
  • Adrenal Glands / metabolism*
  • Child
  • Child, Preschool
  • Chromatography, Liquid / methods*
  • Desoxycorticosterone / blood
  • Female
  • Humans
  • Infant
  • Infant, Newborn
  • Male
  • Progesterone / blood
  • Reference Values
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Steroids / blood*
  • Tandem Mass Spectrometry / methods*
  • Young Adult

Substances

  • Steroids
  • Desoxycorticosterone
  • Progesterone
  • 17-alpha-Hydroxyprogesterone