Validated, Stability Indicating HPTLC Method for the Determination of Mycophenolate Mofetil in Human Plasma

Suraj      P. Verma; Sadaf      J. Gilani; Indu      Bala; D.      P. Maurya; Ozair      Alam; Nadeem      Siddique; Suroor      A. Khan

Abstract

A new high-performance thin-layer chromatographic (HPTLC) method has been established for determination of mycophenolate mofetil in human plasma. Mycophenolate mofetil is used as an important immunosuppressive agent. Chromatographic separation was performed on aluminium plates coated with silica gel 60F254; the mobile phase was a combination of triethylamine buffer (pH 5.3) and acetonitrile in the ratio of 20:80 (v/v) respectively. Densitometric analysis of mycophenolate mofetil was performed at 250 nm. The method was rapid (single-step extraction with acetonitrile), sensitive (limit of quantification 15.4 ng per zone), precise (CV ≤ 3.71 %), accurate (drug recovery 95.08-100.6%), and linear over the range 100-1200 ng per zone. Recovery of mycophenolate mofetil from plasma samples was 95.8 ± 4.5%. The half-life of mycophenolate mofetil in plasma was 20.4 h at 4°C and 17.8 h at 20°C. Mycophenolate mofetil is stable in human plasma for at least two months at -20°C and can tolerate two freeze-thaw cycles with losses < 10%. The method was successfully used to determine therapeutic levels of mycophenolate mofetil.

Keywords: Mycophenolate mofetil, High-performance thin-layer chromatography, Human plasma, Densitometric analysis, Limit of quantification, Liquid chromatographic mass spectrometry, Specificity, Freeze-Thaw Stability, Precision, Accuracy