Running head: TESTOSTERONE MEASUREMENT IMMUNOASSAYS 1 A comparison of salivary testosterone measurement using immunoassays and tandem mass spectrometry

Enzyme immunoassays (EIAs) are widely used to measure salivary testosterone. However, little is known about how accurately different EIAs assess testosterone, partially because estimates across various EIAs differ considerably. We compared testosterone concentrations across EIAs of three commonly used manufacturers (DRG International, Salimetrics, and IBL International) to liquid chromatography tandem mass spectrometry (LC-MS/MS). Relative to EIAs from Salimetrics and IBL International, EIAs supplied by DRG International provided the closest approximation to LC-MS/MS testosterone concentrations with the least measurement error. Additionally, EIAs largely overestimated testosterone in women’s saliva samples with higher testosterone concentrations, relative to men’s. Examining our results and comparing them to existing data revealed that testosterone EIAs had decreased linear correspondence with LC-MS/MS in comparison to cortisol EIAs. Overall, this paper provides researchers with information to better measure testosterone in their research and more accurately compare testosterone measurements across different methods.