# Apparent Hyperandrogenemia Due to Immunoassay Interference Resolved by Liquid Chromatography–Tandem Mass Spectrometry

**Authors:** Dongni Huang, ZhiXuan Guo, JingWen Fan, Yan Zhao, Qi Pan, Lixin Guo

PMC · DOI: 10.1210/jcemcr/luaf140 · 2025-07-02

## TL;DR

A woman's apparently high testosterone levels were found to be a lab error, resolved using a more accurate testing method.

## Contribution

Demonstrates how immunoassay interference can falsely indicate hyperandrogenism, resolved by LC-MS/MS.

## Key findings

- Elevated testosterone levels in a female patient were shown to be an immunoassay artifact.
- LC-MS/MS confirmed normal testosterone levels, resolving diagnostic uncertainty.
- Heterophilic antibodies were identified as the source of the assay interference.

## Abstract

Testosterone is commonly measured using immunoassays in clinical practice; however, these methods are subject to analytical interference in both men and women. In women, due to physiologically lower testosterone levels, even modest elevations may appear pathologic and complicate the diagnostic process. We report a case of a 43-year-old female patient with persistently elevated serum testosterone levels documented across multiple institutions. Despite comprehensive endocrine evaluations and imaging studies, no underlying cause could be identified. Ultimately, liquid chromatography–tandem mass spectrometry (LC-MS/MS) confirmed normal testosterone levels, indicating that the falsely elevated testosterone was due to assay interference. Further analysis using a protein precipitation assay revealed the presence of heterophilic antibodies, which were responsible for the assay interference. This case highlights the importance of considering immunoassay artifacts in the differential diagnosis of unexplained biochemical hyperandrogenism, particularly in the absence of clinical signs of androgen excess.

## Full-text entities

- **Diseases:** hyperandrogenism (MESH:D017588), androgen excess (MESH:D014770), Hyperandrogenemia (MESH:D011085)
- **Chemicals:** Testosterone (MESH:D013739)
- **Species:** Homo sapiens (human, species) [taxon 9606]

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Source: https://tomesphere.com/paper/PMC12214029