Report from the HarmoSter study: different LC-MS/MS androstenedione, DHEAS and testosterone methods compare well; however, unifying calibration is a double-edged sword

• Published in: Clinical chemistry and laboratory medicine Vol. 62; no. 6; pp. 1080 - 1091
• Main Authors: Fanelli, Flaminia, Peitzsch, Mirko, Bruce, Stephen, Cantù, Marco, Temchenko, Anastasia, Mezzullo, Marco, Lindner, Johanna M., Hawley, James M., Ackermans, Mariette T., Van den Ouweland, Jody, Koeppl, Daniel, Nardi, Elena, MacKenzie, Finlay, Binz, Pierre-Alain, Rauh, Manfred, Keevil, Brian G., Vogeser, Michael, Eisenhofer, Graeme, Heijboer, Annemieke C., Pagotto, Uberto
• Format: Journal Article
• Language: English
• Published: Germany De Gruyter 27.05.2024; Walter De Gruyter & Company
• Subjects: Androgens; Androstenedione; Androstenedione - analysis; Androstenedione - blood; Bias; Calibration; Chromatography; Dehydroepiandrosterone; Dehydroepiandrosterone sulfate; Dehydroepiandrosterone Sulfate - analysis; Dehydroepiandrosterone Sulfate - blood; Dehydroepiandrosterone Sulfate - standards; Female; Females; harmonization; Humans; inter-laboratory performance; Laboratories; Liquid chromatography; Liquid Chromatography-Mass Spectrometry - methods; Liquid Chromatography-Mass Spectrometry - standards; liquid chromatography-tandem mass spectrometry; Male; Males; Mass spectrometry; Mass spectroscopy; Middle Aged; Sex hormones; Sulfates; Tandem Mass Spectrometry - methods; Tandem Mass Spectrometry - standards; Testosterone; Testosterone - analysis; Testosterone - blood; Testosterone - standards; liquid chromatography-tandem mass spectrometry; androstenedione; dehydroepiandrosterone sulfate; inter-laboratory performance; testosterone; harmonization
• ISSN: 1434-6621; 1437-4331; 1437-4331
• DOI: 10.1515/cclm-2023-1138

Current liquid chromatography-tandem mass spectrometry (LC-MS/MS) applications for circulating androgen measurements are technically diverse. Previously, variable results have been reported for testosterone. Data are scarce for androstenedione and absent for dehydroepiandrosterone sulfate (DHEAS). We assessed the agreement of androstenedione, DHEAS and testosterone LC-MS/MS measurements among nine European centers and explored benefits of calibration system unification. Androgens were measured twice by laboratory-specific procedures in 78 patient samples and in EQA materials. Results were obtained by and external calibration. Intra- and inter-laboratory performances were valued. Intra-laboratory CVs ranged between 4.2-13.2 % for androstenedione, 1.6-10.8 % for DHEAS, and 4.3-8.7 % and 2.6-7.1 % for female and male testosterone, respectively. Bias and trueness in EQA materials were within ±20 %. Median inter-laboratory CV with vs. external calibration were 12.0 vs. 9.6 % for androstenedione (p<0.001), 7.2 vs. 4.9 % for DHEAS (p<0.001), 6.4 vs. 7.6 % for female testosterone (p<0.001) and 6.8 and 7.4 % for male testosterone (p=0.111). Median bias vs. all laboratory median with and external calibration were -13.3 to 20.5 % and -4.9 to 18.7 % for androstenedione, -10.9 to 4.8 % and -3.4 to 3.5 % for DHEAS, -2.7 to 6.5 % and -11.3 to 6.6 % for testosterone in females, and -7.0 to 8.5 % and -7.5 to 11.8 % for testosterone in males, respectively. Methods showed high intra-laboratory precision but variable bias and trueness. Inter-laboratory agreement was remarkably good. Calibration system unification improved agreement in androstenedione and DHEAS, but not in testosterone measurements. Multiple components, such as commutability of calibrators and EQA materials and internal standard choices, likely contribute to inter-laboratory variability.