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

Eline Jurgens1, Edward J. Knaven1, Eva C.A. Hegeman1, Marjolein W.M. van Gemert1, Judith M.A. Emmen2, Ingrid Willemen2, Yvonne Mulder4, Linda IJsselstijn4, Ben M. de Rooij1,*, Theo H.M. Noij1,3

1Research group Analysis Techniques in Life Sciences, Avans University of Applied Sciences, Breda, the Netherlands. 2Amphia Hospital, Breda, the Netherlands. 3Charles River, ’s-Hertogenbosch, the Netherlands. 4Maasstad Hospital, Rotterdam, the Netherlands.

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-hydroxyprogesterone, 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.

KEYWORDS: steroids, profile, LC-MS/MS, congenital adrenal hyperplasia, saliva.

Read the full text here: https://doi.org/10.17145/jab.19.006

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