Development and Validation of LCMS/MS Methods For Estimation of Bioavailability of Anti-Inflammatory Drugs in Human Plasma

A simple and specific method for determination of bioavailability of Mefenamic acid/ Meloxicam/ Aceclofenac in Human Plasma by liquid chromatography-tandem mass spectrometry operated in positive ionization mode was developed and validated. The LC-MS/MS methods were highly sensitive and suitable for the detection of drug in plasma even in low concentrations. Solid phase extraction methodology was adopted in plasma sample preparation, which provides consistent extraction recovery with minimal endogenous interference and matrix effect. The method involves a rapid solid phase extraction from plasma, simple isocratic chromatographic conditions and mass spectrometric detection in the positive ionization mode using multiple reactions monitoring that enables detection down to low nanogram levels with a short run time. Different parameters such as linearity, range, precision, accuracy, ruggedness and robustness, limit of detection (LOD) and limit of quantification (LOQ) were used for full validation of the method. The results were found to be acceptable as per the guidelines of International Conference on Harmonization (ICH). The method found to be novel, rapid, linear, precise, accurate, robust and rugged and can be successfully applied for the routine analysis of Mefenamic acid / Meloxicam/ Aceclofenac with more sensitivity and covers wider range of quantitation. The method also found to be useful and economical.