Abstract

SIMULTANEOUS ESTIMATION OF FOLIC ACID AND METHOTREXATE IN TABLETS BY HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY

Bhushan B. Sonawane*, Kamlesh K. Saindane, Vijay Kumar M., Sayali U. Warde, Sukhada A. Navaratne and Raman Mohan Singh

ABSTRACT

A new, simple, accurate, fast, economic, and precise high-performance thin-layer chromatographic method was developed and validated for the simultaneous estimation of Folic acid (FA) and Methotrexate (MTX) in a tablet dosage form. The method was developed on silica gel 60 F254 HPTLC plates and the mobile phase was used as ethanol: 1-propanol: 25% ammonia in the ratio of (6:2:2 V/V/V). Densitometric analysis of Folic acid & Methotrexate was performed at the wavelength of 295nm. Well-resolved bands were obtained for Folic acid (RF value of 0.36) and Methotrexate (RF value of 0.49) respectively. The method was developed to ensure appropriate separation of the drug samples and validated as per ICH guidelines. The calibration curve was found to be linear in the concentration range of 50-250 ng/band and 500-1050 ng/band of FA and MTX, respectively. The correlation coefficient is 0.9991 and 0.9990 for FA and MTX, respectively. The method is selective and specific, it was successfully applied in pharmaceutical dosage formulation with potential application in the pharmaceutical analysis of Folic acid and Methotrexate in an individual tablet dosage form.

Keywords: Folic acid, Methotrexate, HPTLC method development, Method Validation.