Development and validation of a serum total testosterone liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay calibrated to NIST SRM 971

► A sensitive LC-MS/MS assay for total testosterone was developed. ► This assay is calibrated to the NIST SRM 971. ► The assay compared well with an immunoassay at concentrations > 60 ng/dl. ► The assay compared well to a LC-MS/MS assay certified by the CDC HoST program.