Clinical determination of 17-hydroxyprogesterone in serum by LC-MS/MS: comparison to Coat-A-Count RIA method

J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Aug 7;840(1):69-74. doi: 10.1016/j.jchromb.2006.04.038. Epub 2006 Jun 5.


17alpha-Hydroxyprogesterone is a metabolic precursor of cortisol; elevated levels of 17alpha-hydroxyprogesterone are indicative of congenital adrenal hyperplasia. Traditional determination by immunoassay is plagued by poor antibody specificity, resulting in significant interferences. This study explores an LC-MS/MS method for the quantitation of 17OHP in serum. Deuterated 17alpha-hydroxyprogesterone was added as internal standard, followed by solid-phase extraction, HPLC separation with a C16-amide reverse-phase column with run time of 7 min, and quantification by MS/MS (positive electrospray ionisation) in the selected reaction monitoring mode (SRM). Transitions monitored were 331>109 for the analyte and 339>113 for the deuterated internal standard. Intra-assay precision (%R.S.D.) was 7.4% at 7 nmol/L, inter-assay precision (%R.S.D.) at 2, 7 and 27 nmol/L was 15.4, 10.0 and 7.9% and accuracy at 0.9 nmol/L was 100%. The method was linear from 0.156 to 80 nmol/L. Lower limit of quantitation was 0.2 nmol/L, providing meaningful data for patients within normal range as well as those with elevated levels.

Publication types

  • Comparative Study
  • Validation Study

MeSH terms

  • 17-alpha-Hydroxyprogesterone / blood*
  • Chromatography, High Pressure Liquid / methods*
  • Humans
  • Mass Spectrometry / methods*
  • Radioimmunoassay / methods*
  • Reference Standards
  • Reproducibility of Results
  • Sensitivity and Specificity


  • 17-alpha-Hydroxyprogesterone