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Ultra-performance liquid chromatography-photodiode array detection (UPLC-PDA) method is fully validated for measuring mycophenolic acid concentrations in human plasma. The plasma spiking with naproxen, an internal standard, was extracted by solid phase extraction. The eluent solution was diluted with milli-Q and injected into an Ultra Performance LC system. Chromatographic separation was performed on reverse phase column, 1.7 μm (100 mm x 2.1 mm I.D.) and using acetronitrile with 2mM Ammonium acetate in Milli-Q water as a mobile phase. The gradient program of mobile phase was performed at flow rate of 0.45 mL/min with 3 minutes of total run time. Photodiode Array (PDA) Detector was selected at 220 nm. The retention times were 1.43 and 1.75 minutes for mycophenolic acid and naproxen, respectively. This method was fully validated and all accepted in terms of selectivity, accuracy, precision, and stability. The linearity of the method revealed a correlation determination of >0.998 within the concentration range of 0.25 20 μg/mL. Recoveries of extraction ranged from 87.390.9% with repeatability. A simple, rapid, and reproducible UPLC/PDA method for determination of mycophenolic acid concentrations in human plasma was performed and fully validated. This method was successfully applied to quantify mycophenolic acid concentrations in therapeutic drug monitoring and bioequivalence study.

Validation of ultra performance liquid chromatography photodiode array method for determination of mycophenolic acid in human plasma and its application