Determination of salivary dehydroepiandrosterone using liquid chromatography--tandem mass spectrometry combined with...

A sensitive liquid chromatography-electrospray ionization-tandem mass spectrometric (LC-ESI-MS-MS) method for the quantification of dehydroepiandrosterone (DHEA) in human saliva has been developed and validated. The saliva was deproteinized with acetonitrile, purified using a Strata-X cartridge, derivatized with the permanently charged reagent, 2-hydrazino-1-methylpyridine (HMP), and subjected to LC-MS-MS. The derivatization with HMP was very effective for increasing the detectability of DHEA in the positive-ESI-MS. Quantification was based on the selected reaction monitoring and androsterone was used as an internal standard. This method allowed the reproducible and accurate quantification of the salivary DHEA using a 200-microl sample and the limit of quantitation for DHEA was 25 pg/ml. No significant matrix effect or change in the measured value by freeze/thaw repetition was observed. The developed method was applied to clinical studies, and produced satisfactory results.