Proton Affinitive Derivatization for Highly Sensitive Determination of Testosterone and Dihydrotestosterone in Saliva Samples by LC-ESI-MS/MS

In this study, researchers used proton affinitive derivatization with picolinic acid and its analogs to enhance the sensitivity of detecting testosterone (T) and 5α-dihydrotestosterone (DHT) in saliva samples via LC-ESI-MS/MS. They converted T and DHT into their picolinate esters and analyzed their chromatographic behavior. The esters showed distinct single peaks, with elution order being picolinate, 3/6-methylpicolinate, and 5-butylpicolinate. While the picolinate and 3/6-methylpicolinate esters were significantly affected by saliva matrix interference, the 5-butylpicolinate esters were only slightly impacted, making them more reliable for sensitive determination of T and DHT in saliva.