Quantitative Profiling of Seven Steroids in Saliva using LC-MS/MS

Abstract

OBJECTIVES: For the diagnosis and treatment of disorders like congenital adrenal hyperplasia, the determination of steroids in saliva might form a more patient-friendly method. Therefore a selective and sensitive LC-MS/MS method was developed for androstenedione, cortisol, cortisone, 11-deoxycortisol, 21-deoxycortisol, 17α-hydroxyprogesterone, and testosterone in saliva. METHODS: Saliva was extracted and concentrated. Steroids were determined by LC-MS/MS, using multiple reaction monitoring upon electrospray ionization. RESULTS: A method was developed to determine seven steroids in saliva in one single LC-MS/MS run. Limits of quantification were 0.05 ng/mL for androstenedione, 0.25 ng/mL for cortisol, 0.50 ng/mL for cortisone, 0.125 ng/mL for 11-deoxycortisol, 0.125 ng/mL for 21-deoxycortisol, 0.125 ng/mL for 17α-hy-droxyprogesterone, and 0.05 ng/mL for testosterone. The within-run repeatability was < 5% and the within-laboratory precision was < 5% for all analytes except androstenedione, where the saliva used in the validation study contained androstenedione levels below the LOQ. CONCLUSIONS: A sensitive and selective method was developed for the determination of a profile of seven steroids in saliva in one analytical run.