Ensuring quality in 17OHP mass spectrometry measurement: an international study assessing isomeric steroid interference

Chung Shun Ho, Kirsten Hoad, Brian R. Cooke, Trisha Andersen, Peter Graham, Sjoerd A.A. van den Berg, Michaela F. Hartmann, Clara W.S. Lo, Tze Ping Loh, Yolanda B. de Rijke, Bertrand D. van Zelst, Stefan A. Wudy, Rosita Zakaria, Ronda F. Greaves*

*Corresponding author for this work

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Interference from isomeric steroids is a potential cause of disparity between mass spectrometry-based 17-hydroxyprogesterone (17OHP) results. We aimed to assess the proficiency of mass spectrometry laboratories to report 17OHP in the presence of known isomeric steroids. 


A series of five samples were prepared using a previously demonstrated commutable approach. These samples included a control (spiked to 15.0 nmol/L 17OHP) and four challenge samples further enriched with equimolar concentrations of 17OHP isomers (11α-hydroxyprogesterone, 11β-hydroxyprogesterone, 16α-hydroxyprogesterone or 21-hydroxyprogesterone). These samples were distributed to 38 participating laboratories that reported serum 17OHP results using mass spectrometry in two external quality assurance programs. The result for each challenge sample was compared to the control sample submitted by each participant. 


Twenty-six laboratories (68 % of distribution) across three continents returned results. Twenty-five laboratories used liquid chromatography-tandem mass spectrometry (LC-MS/MS), and one used gas chromatography-tandem mass spectrometry to measure 17OHP. The all-method median of the control sample was 14.3 nmol/L, ranging from 12.4 to 17.6 nmol/L. One laboratory had results that approached the lower limit of tolerance (minus 17.7 % of the control sample), suggesting the isomeric steroid caused an irregular result. 


Most participating laboratories demonstrated their ability to reliably measure 17OHP in the presence of the four clinically relevant isomeric steroids. The performance of the 12 (32 %) laboratories that did not engage in this activity remains unclear. We recommend that all laboratories offering LC-MS/MS analysis of 17OHP in serum, plasma, or dried bloodspots determine that the isomeric steroids are appropriately separated.

Original languageEnglish
Pages (from-to)911-918
Number of pages8
JournalClinical Chemistry and Laboratory Medicine
Issue number5
Publication statusPublished - 1 Apr 2024

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