Analysis of testosterone in female and juvenile plasma samples for research requires an analytically sensitive method with a limit of quantitation of at least 10 pg/mL. Liquid chromatography coupled with mass spectrometry (LC/MS), an analytically sensitive and selective technique, is widely accepted for testosterone analysis in complex matrices such as human serum or plasma.

The analysis of testosterone in human serum for clinical research is currently performed using many different sample preparation procedures. In this note, we evaluated a Prelude SPLC system coupled to a TSQ Endura MS (Figure 1) in the analysis of testosterone using two sample preparation methods: online sample cleanup and offline sample cleanup. Each method uses specific chromatography, but both share the same mass spectrometer settings and parameters.

To describe an accurate and precise high-throughput analytical technique for the analysis of testosterone, androstenedione, and 17-hydroxy progesterone utilizing micro-scale solid phase extraction (SPE), followed by liquid chromatography coupled with triple quadrupole mass spectrometry (LC-MS/MS). For research use only. Not for use in diagnostic procedures.

A Thermo Scientific HPLC system coupled with the Thermo Scientific TSQ Vantage mass spectrometer are applied for the rapid determination of testosterone. The separation was performed on a Thermo Scientific Accucore C8 HPLC column, with a cycle time of 1.5 minutes.

The Thermo Scientific HPLC system is applied for the determination of testosterones. The separation is performed on a Thermo Scientific Accucore RP-MS HPLC column, with UV detection at 254 nm.