Abstract

METHOD DEVELOPMENT AND VALIDATION OF METHOTREXATE DRUG BY RP-HPLC

A Santosh kumar Sreevatsav*, N. Shiva Prasad, D.Akhilesh Kumar,
Sanjay Kumar M, M.Satish Kumar

ABSTRACT

A simple and reproducible method was developed for Methotrexate by Reverse phase high performance liquid chromatography (RP-HPLC). Methotrexate was separated on C-18 column (1000mm x 6mm x 5μ), using potassium di hydrogen orthophosphate at the UV detection of 303nm. Isocratic elution of Acetonitrile and buffer as mobile phase with various ratios and flow rates. Eventually 92:8 v/v buffer and acetonitrile was being set with the flow rate of 1.4 ml/min. The statistical validation parameter such as selectivity (or) specificity, linearity, precision,accuracy, robustness were checked. The recovery and assay studies of method were within 97% to 102%. Indicates that the proposed method can be adoptable for Quality control analysis of method.

Keywords: Methotrexate, RP-HPLC, Validation, acetonitrile