title: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for simultaneous measurement of salivary testosterone and cortisol in healthy men for utilization in the diagnosis of late-onset hypogonadism in males.
abstract: It is well known that late-onset hypogonadism in males can cause a variety of symptoms, and the differential diagnosis is relatively difficult, including psychological disorders, stress, and mood disturbances. The level of serum cortisol can be measured to reflect a patient's level of stress. Salivary hormones facilitate the evaluation of physiological hormonal actions based on free hormone assay. For the simultaneous measurement of testosterone and cortisol levels in saliva, we validate a sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay. Concerning accuracy and precision, the lower limit of quantification of salivary testosterone and cortisol were established as 5 and 10 pg/mL, respectively. Testosterone and cortisol in saliva is stable for 2 days, 14 days, and 28 days at room temperature, refrigeration and frozen, respectively. Freezing and thawing for 3 cycles and stimulation of salivation with gum chewing do not alter the measured values of testosterone and cortisol. Total, bioavailable, and free serum testosterone showed slight diurnal changes, but total and bioavailable serum cortisol showed marked diurnal changes. Salivary testosterone levels negatively correlate with age, regardless of the time of saliva collection (r=0.64, p<0.05). However, there is no relationship between salivary cortisol and age (r=0033, p>0.05). LC-MS/MS allows rapid, simultaneous, sensitive, and accurate quantification of testosterone and cortisol in saliva for the diagnosis late-onset hypogonadism or other hormone related disease.
authors.affiliation: Department of Integrative Cancer Therapy and Urology, Kanazawa University Graduate School of Medical Science, Ishikawa, Japan.
authors.forename: Futoshi
authors.initials: F
authors.lastname: Matsui
authors.forename: Eitetsu
authors.initials: E
authors.lastname: Koh
authors.forename: Kenrou
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Sugimoto
authors.forename: Ho-Su
authors.initials: HS
authors.lastname: Sin
authors.forename: Yuji
authors.initials: Y
authors.lastname: Maeda
authors.forename: Seijiro
authors.initials: S
authors.lastname: Honma
authors.forename: Mikio
authors.initials: M
authors.lastname: Namiki
mesh_terms: D000328:Adult; D017668:Age of Onset; D000368:Aged; D000375:Aging; D002638:Chewing Gum; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002940:Circadian Rhythm; D003863:Depression / Q000175:diagnosis*; D003937:Diagnosis, Differential; D006801:Humans; D006854:Hydrocortisone / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism*; D007006:Hypogonadism / Q000175:diagnosis*; D057230:Limit of Detection; D008297:Male; D008875:Middle Aged; D019964:Mood Disorders / Q000175:diagnosis*; D015203:Reproducibility of Results; D012463:Saliva / Q000378:metabolism*; D012738:Sex Hormone-Binding Globulin / Q000378:metabolism; D013048:Specimen Handling / Q000379:methods; D013315:Stress, Psychological / Q000175:diagnosis*; D053719:Tandem Mass Spectrometry / Q000379:methods; D013739:Testosterone / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism*; D013997:Time Factors
chemical_list: D002638:Chewing Gum; D012738:Sex Hormone-Binding Globulin; D013739:Testosterone; D006854:Hydrocortisone