Determination of prednisolone in human adipose tissue incubation medium using LC-MS/MS to support the measurement of 11 beta-hydroxysteroid dehydrogenase activity

作者：Ding X; Rose M J^*; McCaffery I; Rossi J; Paweletz K; Hale C; Emery M; James C A

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2009, 877(13): 1394-1401.

DOI：10.1016/j.jchromb.2008.09.025

• Prednisolone
• 11 beta-Hydroxysteroid dehydrogenase type 1
• LC-MS/MS
• mu Elute 96-well SPE
• Human adipose tissue
• Biomarker

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 11 beta-hydroxysteroid dehydrogenase in human adipose tissue. After incubation, samples (80 mu L) were extracted using Oasis HLB mu Elute 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 TurbolonSpray (R) 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 11 beta-hydroxysteroid dehydrogenase type 1 (11 beta-HSD1) enzyme activity in human adipose tissue.

• 出版日期2009-5-1