CHEMOMETRIC ASSISTED NEW STABILITY INDICATING NPHPLC METHOD DEVELOPMENT AND VALIDATION OF CLAVULANIC ACID , AMOXCILLNE AND LACTOBACILLUS IN COMBINED DOSAGE FORM

Abstract

A new chemometric assisted by high-performance liquid chromatography (HPLC) with photodiode array (PDA)
detection was implemented for the simultaneous determination of tablet dosage form. Two chemometric
calibration techniques, principle component analysis (PCA) and partial least squares (PLS) were applied to the
peak area at 246nm of PDA detector responses. Chromatographic separation of Amoxicillin, Clavulanic acid
and Lactobacillus was achieved on Waters Alliance-e2695, by using Chiral Cell ODH 150x4.6mm, 5µ column
and the mobile phase containing Hexane: THF: Acetic acid in the ratio of 96.5:3:0.5% v/v. The flow rate was
1.0 ml/min; detection was carried out by absorption at 246nm using a photodiode array detector at ambient
temperature. The number of theoretical plates and tailing factor for Amoxicillin, Clavulanic acid and
Lactobacillus were NLT 2000 and should not more than 2 respectively. % 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 Amoxicillin, Clavulanic acid 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 thequantitative analysis of Amoxicillin,
Clavulanic acid and Lactobacillus in pharmaceutical tablet dosage form