CHEMOMETRIC ASSISTED NP-HPLC METHOD FOR THE DEVELOPMENT AND VALIDATION OF NORFLOXACIN AND LACTOBACILLUS

Abstract

A new chemometric carried out by High Performance Liquid Chromatography (HPLC) through photodiode array
(PDA) detection was executed for the simultaneous estimation of tablet dosage form. Two chemometric calibration
techniques, principle component analysis (PCA) and partial least squares (PLS) were applied to the peak area at 242
nm of PDA detector responses. Chromatographic separation of Norfloxacin and Lactobacillus was achieved on
Waters Alliance-e2695, by using Chiral Cell ODH 150 × 4.6 mm, 5µ column and the mobile phase containing
Hexane : IPA : Ethanol in the ratio of 20:40:40% v/v. The flow rate was 1.0 ml/min; detection was carried out by
absorption at 242 nm using a photodiode array detector at ambient temperature. The number of theoretical plates and
tailing factor for Norfloxacin and Lactobacillus were NLT 2000 and should not more than 2 respectively. The %
Relative standard deviation of peak areas of all measurements always less than 2.0. The proposed method was
validated according to ICH guidelines. The method was found to be simple, economical, suitable, precise, accurate
& robust method for quantitative analysis of Norfloxacin and Lactobacillus and study of its stability. The
‘UNSCRAMBLER (camo)’software was used for the numerical calculations. All of the two-chemometric analysis
methods in this study can be satisfactorily applied for the quantitative analysis of Norfloxacin and Lactobacillus in
pharmaceutical tablet dosage form