Candidate reference measurement procedure based on isotope dilution-two dimensional-liquid chromatography-tandem mass spectrometry for the quantification of androstenedione in human serum and plasma

Ott M, Bauland F, Fischer V, Köppl D, Gaudl A, Geistanger A, Ceglarek U, Rauh M, Taibon J (2025)

Publication Type: Journal article

Publication year: 2025

Journal

Clinical Chemistry and Laboratory Medicine Walter de Gruyter

DOI: 10.1515/cclm-2024-1135

Abstract

Objectives: Androstenedione (ASD) is a biomarker used in the diagnosis of hyperandrogenism and adrenal hyperplasia. Therefore, accurate measurement of serumASD is pivotal in clinical settings. We aimed to develop a novel highly selective reference measurement procedure (RMP) based on isotope dilution-two dimensional-liquid chromatographytandem mass spectrometry (ID-2D-LC-MS/MS) for the quantification of ASD in human serum/plasma. Methods: To achieve high selectivity and sensitivity, a twodimensional heart-cut LC approach for LC-MS/MS and a supported liquid extraction sample preparation protocol were employed. Matrix effects were evaluated through a post-column infusion experiment and comparison of standard line slopes. Precision and accuracy were determined via a multi-day validation experiment. Reproducibility was assessed by comparing results from two independent laboratories, and measurement uncertainty (MU) was evaluated in compliance with current guidelines. Results: Our novel RMP proved effective for measuring ASD concentrations ranging from 0.00800 ng/mL to 12.0 ng/mL (0.0279 nmol/L to 41.9 nmol/L) and demonstrated matrixindependence. The relative mean bias varied between 0.6 and 2.2 %across different matrices and concentration levels. Intermediate precision was observed to be between 1.2 and 1.9 %. The expanded measurement uncertainty for ASD target value assignment ranged from 1.7 to 2.5 %, irrespective of sample concentration. Equivalence to the JCTLMlisted RMP was evaluated through a method comparison study, revealing a high degree of agreement (r≥0.997). Additionally, by comparing results from two independent laboratories, the transferability and reproducibility of the RMP were confirmed. Conclusions: This isotope dilution two-dimensional LC-MS/ MS method can be used for the evaluation and standardization of routine assays and for measuring individual patient samples, ensuring traceability.

Authors with CRIS profile

Manfred Rauh Department of Paediatrics and Adolescent Medicine

Involved external institutions

Roche Diagnostics GmbH DE Germany (DE) Chrestos Concept GmbH & Co. KG DE Germany (DE) Universität Leipzig DE Germany (DE)

How to cite

APA:

Ott, M., Bauland, F., Fischer, V., Köppl, D., Gaudl, A., Geistanger, A.,... Taibon, J. (2025). Candidate reference measurement procedure based on isotope dilution-two dimensional-liquid chromatography-tandem mass spectrometry for the quantification of androstenedione in human serum and plasma. Clinical Chemistry and Laboratory Medicine. https://doi.org/10.1515/cclm-2024-1135

MLA:

Ott, Myriam, et al. "Candidate reference measurement procedure based on isotope dilution-two dimensional-liquid chromatography-tandem mass spectrometry for the quantification of androstenedione in human serum and plasma." Clinical Chemistry and Laboratory Medicine (2025).

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