Development and Validation of RP-HPLC Method for Estimation of Methotrexate Drug Intercalated in Mg-Al Layered Double Hydroxide Nanoparticles

Abstract

In the present study, pristine magnesium aluminium layered double hydroxide (Mg-Al LDH, sample A) nanoparticles were synthesized by simple co-precipitation technique followed by intercalation of methotrexate drug (MTX), their detailed physicochemical characterization and development of RP-HPLC method for estimation of MTX intercalated in Mg-Al LDH nanoparticles (sample B). The PXRD analysis showed the basal spacing of 003 plane corresponding to sample A and sample B to be 8.63 Å and 11.39 Å, respectively. Particle size distributions were measured to be 56-75 nm (D50=66.15 nm) in case of sample A and 75-100 nm (D50=87.41 nm) for sample B. Simple, accurate RP-HPLC method was developed for estimation of the MTX drug from sample B, on Agilent HPLC 1200 system with UV detector on L1 column. 0.02 M potassium dihydrogen phosphate solution (pH 3.0) and acetonitrile (85:15 v/v) was used as mobile phase in an isocratic elution mode at a flow rate of 1.0 mL/min at 35°C with a load of 10 μL at 307 nm. The retention time for methotrexate was at 4.44 min. The method was validated as per ICH Q2 (R1) guidelines. The method was found to be linear over a concentration range of 10-100 μg/mL.