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Accurate Method of HPLC-Ms/Ms Determination of Mycophenolic Acid in Human Plasma

Khokhlov AL, Yaichkov II, Shitov LN, Dzhurko YA, Shitova AM, Ryska M, Kubeš V, Shabrov VN and Miroshnikov AE

A simple, accurate and precise method was developed using HPLC-MS/MS to quantify mycophenolic acid in human plasma. Mycophenolic acid-D3 was used as internal standard. Sample preparation involved protein precipitation. Chromatographic analysis was performed on a Phenomenex Kinetex C18 (30 mm × 4.6 mm, 2.6 μm) column using acetonitrile-water as the mobile phase. Detection in negative ionization mode was used to quantify mycophenolic acid by multiple reaction monitoring. The calibration curve was linear in the range of 0.5-30 μg/mL, with intra-and inter-batch values for accuracy and precision (coefficient of variation) ranging from 99.76 to 111.38% and from 2.54 to 9.01%, respectively. The method was successfully applied to conduct a pharmacokinetic study of 360 mg coated tablet "Myfortic" in 48 healthy participants; 768 blood samples were analysed.