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  • Title: Determination of prednisolone in human adipose tissue incubation medium using LC-MS/MS to support the measurement of 11beta-hydroxysteroid dehydrogenase activity.
    Author: Ding X, Rose MJ, McCaffery I, Rossi J, Paweletz K, Hale C, Emery M, James CA.
    Journal: J Chromatogr B Analyt Technol Biomed Life Sci; 2009 May 01; 877(13):1394-401. PubMed ID: 18945652.
    Abstract:
    A method for the determination of prednisolone in human adipose tissue incubation medium has been developed, validated and used to support studies designed to measure the activity of 11beta-hydroxysteroid dehydrogenase in human adipose tissue. After incubation, samples (80 microL) were extracted using Oasis HLB microElute SPE plates and the resulting extracts were analyzed using reversed-phase chromatography coupled to an Applied Biosystems Sciex PE API-4000 mass spectrometer with a TurboIonSpray interface (400 degrees C). The method was validated over the calibration range of 0.5-100 ng/mL. Intraday precision and accuracy were 6.1% R.S.D. or less and within 6.3%, respectively. Interday precision and accuracy were 4.2% R.S.D. or less and within 3.6%, respectively. Extraction recovery of prednisolone was greater than 84% over the range of low to high quality control sample concentrations. The validated assay was used to support studies designed to estimate ex vivo 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) enzyme activity in human adipose tissue.
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