Development and Validation of UV-Visible Spectrophotometric Method for the Analysis of Methotrexate in Pharmaceutical Formulations

A simple, sensitive, selective rapid spectrophotometric method has been developed for the determination of post synaptic α1- Adrino receptor antagonist Methotrexate in pure form and pharmaceutical formulations based on the comparative method of analysis and recovery studies like reductive coupling reaction and diazotization with1,10-PT,2,2-BP, FC reagent, NaNO2/HCl and k3FeCN6/FeCl3 at P H_4.0 which is extractable at 430 nm,480nm,680nm,440nm & 680nm. Beer’s law is obeyed in the concentration ranges 0.5 -3 µg/ml,1-6 µg/ml,0.4-2 µg/ml1-4 µg/ml and 0.1 -0.5 µg/ml . The developed method was applied directly and easily for the analysis of the Pharmaceutical formulations and R.S.D was found to be 0.46%,1.5%,0.6%,0.88% and 0.89% and Recovery 99.92 ± 0.12 , 99.19 ± 0.12, 99.81 ± 0.12 100.03 ± 0.72 and 99.95 ± 0.15 respectively. The method was completely validated and proven to be rugged. The interferences of the other ingredients and excipients were not observed. The repeatability and the performance of the proved method were established by point and internal hypothesis and through recovery studies