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J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Jul 15;870(2):251-4. doi: 10.1016/j.jchromb.2008.06.020. Epub 2008 Jun 20.

Determination of mycophenolic acid glucuronide in microsomal incubations using high performance liquid chromatography-tandem mass spectrometry.

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Department of Pharmacy Practice, College of Pharmacy, University of Florida, PO Box 100486, Gainesville, FL 32610, USA.


A sensitive and specific HPLC-MS/MS method was developed for the analysis of mycophenolic acid glucuronide (MPAG) in incubations with human liver microsomes. Incubation samples were processed by protein precipitation with acetonitrile. MPAG and the internal standard phenolphthalein glucuronide were chromatographed on a C18 Synergi Fusion-RP column (100 mm x 2 mm, 4 microm) using gradient elution with a mixture of 1mM acetic acid in deionized water and 1mM acetic acid in acetonitrile at a flow rate of 0.22 mL/min. The mass spectrometer was operated with negative electrospray ionization and analysis was carried out in the single reaction monitoring (SRM) mode using the mass transitions of m/z 495-->319 and m/z 493-->175 for MPAG and phenolphthalein glucuronide, respectively. The MPAG calibration curve was linear over the concentration range of 1.0-20 microM. The within-day and between-day relative standard deviations ranged from 1.1 to 7.9% and accuracy was within 8%. The simple and reproducible method is suitable for measuring mycophenolic acid glucuronidation in microsomal incubations.

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