Bioanalytical method development and validation of mefenamic acid in human plasma by liquid chromatography coupled with tandem mass spectrometry

In this present study the bioanalytical method development and validation of an LC-MS/MS method in human plasma for the quantification of mefenamic acid is described .The method carried out by using liquid-liquid extraction as diclofenac is utilised as an internal standard (IS). Chromatographic separation was carried out on a Hypersil Gold C-18 (100 mm x 4.8 mm, 5 µm) column, using a mobile phase composed of methanol and ammonium acetate buffer in a 70:30 ratio. The flow rate was set at 1 mL/min, and the total runtime was 3 minutes. The analyte was subjected to LC-MS/MS using positive ionization source. The standard curve was linear from 81.94 ng/ml to 10246.69 ng/ml. This assay allows quantification of mefenamic acid at lower concentrations in human plasma. The observed mean recovery value was found to be 68.03% for Mefenamic acid and 94.74% for Diclofenac respectively. From this observed results it was concluded that the developed method was more economic, precise, accurate and lesser time utility for the routine analysis.