Rapid RP-HPLC Estimation of Methotrexate in Bulk, Pharmaceutical Preparation and in Spiked Plasma samples.

Abstract

A simple, sensitive and selective RP-HPLC method with UV detection for the estimation of Methotrexate in pharmaceutical formulation and in spiked plasma developed and validate in present work. Chromatographic separation of drug is performed with a 250X4.6 mm, 5µm diameter particles RP C-18 column and the mobile phase consisted of a mixture of methanol and water (80:20, v/v), containing 0.1% HPLC grade glacial acetic acid for the adjustment of pH to 4.5. Isocratic elution at a flow rate of 1 ml/min with UV detection at 256 nm at ambient temperature is used in this method. The proposed RP-HPLC method is successfully applied for the determination of MTX in pharmaceutical preparation and spiked plasma samples. The validation studies are carried out and it’s fulfilling ICH requirements. The method is found to be specific, linear, precise (including both intra- and inter- day precision), accurate and robust. This proposed method may represent a valuable aid in the laboratory monitoring of the toxicity of anticancer chemotherapy.

Keywords: Methotrexate, RP-HPLC method, Spiked plasma samples, Estimation, Toxicity monitoring