TY - JOUR T1 - Simultaneous determination of testosterone, cortisol, and dehydroepiandrosterone in saliva by stable isotope dilution on-line in-tube solid-phase microextraction coupled with liquid chromatography-tandem mass spectrometry. AU - Kataoka,Hiroyuki, AU - Ehara,Kentaro, AU - Yasuhara,Rie, AU - Saito,Keita, Y1 - 2012/10/13/ PY - 2012/08/27/received PY - 2012/10/02/accepted PY - 2012/09/30/revised PY - 2012/10/16/entrez PY - 2012/10/16/pubmed PY - 2013/6/26/medline SP - 331 EP - 40 JF - Analytical and bioanalytical chemistry JO - Anal Bioanal Chem VL - 405 IS - 1 N2 - We have developed a simple and sensitive method for the simultaneous determination of testosterone (TES), cortisol (CRT), and dehydroepiandrosterone (DHEA) in saliva by automated online in-tube solid-phase microextraction (SPME) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using a Discovery HS F5 column. The optimum in-tube SPME conditions were 25 draw/eject cycles of 40 μL of sample at a flow rate of 200 μL/min using a Supel-Q PLOT capillary column as an extraction device. The extracted compounds were easily desorbed from the capillary by passage of the mobile phase, and no carryover was observed. The in-tube SPME LC-MS/MS method showed good linearity with correlation coefficients r ≥ 0.9998 for TES, CRT, and DHEA using their respective stable isotope-labeled internal standards. The intra-day and inter-day precisions (relative standard deviations) were below 4.9 and 8.5 % (n = 5), respectively. This method was successfully utilized to analyze TES, CRT, and DHEA in saliva samples without any other pretreatment or interference peaks, and the quantification limits (S/N = 10) of TES, CRT and DHEA were about 0.01, 0.03 and 0.29 ng/mL saliva, respectively. The recoveries of these compounds spiked into saliva samples were each above 94%. This method was applied to analyze changes in salivary TES, CRT, and DHEA levels resulting from stress and fatigue load. SN - 1618-2650 UR - https://www.unboundmedicine.com/medline/citation/23064711/Simultaneous_determination_of_testosterone_cortisol_and_dehydroepiandrosterone_in_saliva_by_stable_isotope_dilution_on_line_in_tube_solid_phase_microextraction_coupled_with_liquid_chromatography_tandem_mass_spectrometry_ L2 - https://dx.doi.org/10.1007/s00216-012-6479-4 DB - PRIME DP - Unbound Medicine ER -