Report from the HarmoSter study: Impact of calibration on comparability of LC-MS/MS measurement of circulating cortisol, 17OH-progesterone and aldosterone

Flaminia Fanelli, Marco Cantù, Anastasia Temchenko, Marco Mezzullo, Johanna M. Lindner, Mirko Peitzsch, James M. Hawley, Stephen Bruce, Pierre-Alain Binz, Mariette T. Ackermans, Annemieke C. Heijboer, Jody van den Ouweland, Daniel Koeppl, Elena Nardi, Finlay Mackenzie, Manfred Rauh, Graeme Eisenhofer, Brian G. Keevil, Michael Vogeser, Uberto Pagotto

Research output: Contribution to journalArticle*Academicpeer-review


Objectives: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is recommended for measuring circulating steroids. However, assays display technical heterogeneity. So far, reproducibility of corticosteroid LC-MS/MS measurements has received scant attention. The aim of the study was to compare LC-MS/MS measurements of cortisol, 17OH-progesterone and aldosterone from nine European centers and assess performance according to external quality assessment (EQA) materials and calibration. Methods: Seventy-eight patient samples, EQA materials and two commercial calibration sets were measured twice by laboratory-specific procedures. Results were obtained by in-house (CAL1) and external calibrations (CAL2 and CAL3). We evaluated intra and inter-laboratory imprecision, correlation and agreement in patient samples, and trueness, bias and commutability in EQA materials. Results: Using CAL1, intra-laboratory CVs ranged between 2.8-7.4%, 4.4-18.0% and 5.2-22.2%, for cortisol, 17OH-progesterone and aldosterone, respectively. Trueness and bias in EQA materials were mostly acceptable, however, inappropriate commutability and target value assignment were highlighted in some cases. CAL2 showed suboptimal accuracy. Median inter-laboratory CVs for cortisol, 17OH-progesterone and aldosterone were 4.9, 11.8 and 13.8% with CAL1 and 3.6, 10.3 and 8.6% with CAL3 (all p<0.001), respectively. Using CAL1, median bias vs. all laboratory-medians ranged from -6.6 to 6.9%, -17.2 to 7.8% and -12.0 to 16.8% for cortisol, 17OH-progesterone and aldosterone, respectively. Regression lines significantly deviated from the best fit for most laboratories. Using CAL3 improved cortisol and 17OH-progesterone between-method bias and correlation. Conclusions: Intra-laboratory imprecision and performance with EQA materials were variable. Inter-laboratory performance was mostly within specifications. Although residual variability persists, adopting common traceable calibrators and RMP-determined EQA materials is beneficial for standardization of LC-MS/MS steroid measurements.

Original languageEnglish
Pages (from-to)726-739
Number of pages14
JournalClinical Chemistry and Laboratory Medicine
Issue number5
Early online date2022
Publication statusPublished - 1 Apr 2022


  • 17OH-progesterone
  • aldosterone
  • calibration
  • cortisol
  • external quality control
  • harmonization
  • inter-laboratory performance
  • liquid chromatography-tandem mass spectrometry
  • method comparison
  • steroid hormones

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