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Measurement of Urinary Free Cortisol and Cortisone by LC-MS/MS

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Clinical Applications of Mass Spectrometry in Biomolecular Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2546))

Abstract

Monitoring urinary free cortisol (UFC) excretion helps assess adrenal function and is used to screen for endogenous Cushing’s syndrome caused by an adrenal or pituitary tumor. While serum cortisol levels fluctuate in response to time of day, stress, and concentrations of cortisol-binding globulin (CBG), a 24-h urine collection measures the cortisol produced over the entire day and does not suffer from as much variability as a serum measurement.

We describe here a method of measurement of urinary free cortisol (UFC) and cortisone using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Urine samples, combined with stable isotope-labeled internal standards, are extracted by liquid-liquid extraction using ethyl acetate and hexane. An API 5500 mass spectrometer operated in positive atmospheric pressure chemical ionization (APCI) mode is used for detection.

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Acknowledgments

We thank the ARUP Institute for Clinical and Experimental Pathology® for supporting this project.

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Correspondence to Julie A. Ray .

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Ray, J.A., Kish-Trier, E., Johnson, L.M. (2022). Measurement of Urinary Free Cortisol and Cortisone by LC-MS/MS. In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Biomolecular Analysis. Methods in Molecular Biology, vol 2546. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2565-1_11

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  • DOI: https://doi.org/10.1007/978-1-0716-2565-1_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2564-4

  • Online ISBN: 978-1-0716-2565-1

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