The contribution of liquid chromatography coupled to tandem mass spectrometry (LC MS/MS) in the analysis of testosterone

The combination of liquid chromatography coupled to tandem mass spectrometry (LC MS/MS) has significantly improved testosterone analysis, offering an analytical method with high sensitivity, specificity and also reproducibility suitable for many applications. This approach is particularly relevant in clinical diagnostics, sports science and the control of substances in dietary supplements. The integration of LC-MS/MS enables accurate and reliable quantification of testosterone concentrations, alleviating the limitations inherent in conventional immunoassays. Testosterone is a key hormone involved in the regulation of numerous physiological processes in both men and women. Accurate analysis of this hormone is crucial for the diagnosis of endocrine dysfunctions, such as polycystic ovary syndrome and hypogonadism, as well as for routine testing. Testosterone concentrations have traditionally been assessed using serum or plasma samples, an approach that presents challenges in terms of collection, storage and transport, particularly in regions with limited resources. Measurement of testosterone using immunoassays has proved imprecise, particularly at low concentrations, due to interference from cross-reactivity with structurally related steroid hormones or synthetic derivatives. Immunoassays are widely used in clinical laboratories for the measurement of testosterone in serum/plasma, although second-generation assays have improved considerably, liquid chromatography-tandem mass spectrometry, LC-MS/MS, has been recommended as a better alternative, due to its high specificity and accuracy and its ability to measure over a wide range of concentrations. The use of dried blood drops (DBS) has emerged as an optimized strategy for hormone analysis, offering significant advantages in terms of sample stability, ease of collection and streamlining of logistical processes. The aim of this study is to optimize the contribution of liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in testosterone analysis, to ensure validity and accuracy comparable to conventional serum methods.