Harmonisation of serum dihydrotestosterone analysis: establishment of an external quality assurance program

Ronda F. Greaves 1 , 2 , Lisa Jolly 3 , Michaela F. Hartmann 4 , Chung Shun Ho 5 , Richard K.T. Kam 5 , John Joseph 6 , Conchita Boyder 6  and Stefan A. Wudy 4
  • 1 School of Health and Biomedical Sciences, RMIT University, PO Box 71, Bundoora, Victoria 3083, Australia, Phone: +61 3 9925 7080
  • 2 Centre for Hormone Research, Murdoch Children’s Research Institute, Victoria, Australia
  • 3 RCPA Quality Assurance Programs, Chemical Pathology, Adelaide, South Australia, Australia
  • 4 Steroid Research and Mass Spectrometry Unit, Laboratory for Translational Hormone Analytics, Pediatric Endocrinology, Justus Liebig University, Giessen, Germany
  • 5 Biomedical Mass Spectrometry Unit, Department of Chemical Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, New Territories, Hong Kong SAR
  • 6 PathWest Laboratory Medicine, QE2 Medical Centre, Perth Western Australia, Australia
Ronda F. Greaves, Lisa Jolly, Michaela F. Hartmann, Chung Shun Ho, Richard K.T. Kam, John Joseph, Conchita Boyder and Stefan A. Wudy

Abstract

Background:

Serum dihydrotestosterone (DHT) is an important analyte for the clinical assessment of disorders of sex development. It is also reportedly a difficult analyte to measure. Currently, there are significant gaps in the standardisation of this analyte, including no external quality assurance (EQA) program available worldwide to allow for peer review performance of DHT. We therefore proposed to establish a pilot EQA program for serum DHT.

Methods:

DHT was assessed in the 2015 Royal College of Pathologists of Australasia Quality Assurance Programs’ Endocrine program material. The material’s target (i.e. “true”) values were established using a measurement procedure based on isotope dilution gas chromatography (GC) tandem mass spectrometry (MS/MS). DHT calibrator values were based on weighed values of pure DHT material (>97.5% purity) from Sigma. The allowable limits of performance (ALP) were established as ±0.1 up to 0.5 nmol/L and ±15% for targets >0.5 nmol/L.

Results:

Target values for the six levels of RCPAQAP material for DHT ranged from 0.02 to 0.43 nmol/L (0.01–0.12 ng/mL). The material demonstrated linearity across the six levels. There were seven participating laboratories for this pilot study. Results of the liquid chromatography (LC) MS/MS methods were within the ALP; whereas the results from the immunoassay methods were consistently higher than the target values and outside the ALP.

Conclusions:

This report provides the first peer comparison of serum DHT measured by mass spectrometry (MS) and immunoassay laboratories. Establishment of this program provides one of the pillars to achieve method harmonisation. This supports accurate clinical decisions where DHT measurement is required.

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