DEVELOPMENT AND VALIDATION OF HPTLC METHOD FOR SIMULTANEOUS DETERMINATION OF PAMABROM AND PARACETAMOL IN SYNTHETIC MIXTURE

Abstract

A new, simple, precise, accurate and selective high performance thin-layer chromatographic (HPTLC) method has been developed and validated for the simultaneous determination of pamabrom and paracetamol in a synthetic mixture. Chromatographic separation was carried out on Merck TLC aluminium sheets of silica gel 60F254 using Chloroform: Acetonitrile (5.0: 5.0 % v/v) as mobile phase followed by densitometric analysis at 277 nm. This system was found to give compact spots for pamabrom (Rf value of 0.34 ± 0.004) and paracetamol (Rf value of 0.56 ± 0.004). The method was validated in terms of linearity, accuracy, precision, limit of detection, limit of quantification and specificity in accordance with International Conference on Harmonization (ICH) guidelines. The calibration curve was found to be linear between 100 to 350 and 1300 to 4550 ng/spot for pamabrom and paracetamol, respectively with significantly high value of correlation coefficient (r² > 0.99). The limits of detection and quantitation were found to be 7.65 and 23.17 ng/spot, respectively for pamabrom and 52.63 and 159.48 ng/spot, respectively for paracetamol. The proposed method was found to be accurate, precise, reproducible, specific and sensitive and can be applicable for the simultaneous determination of pamabrom and paracetamol in Synthetic mixture.