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Abstract

DEVELOPMENT AND VALIDATION OF A STABILITY INDICATING RP-HPLC METHOD FOR ASSAY DETERMINATION OF METHYLPARABEN, PROPYLPARABEN, POTASSIUM SORBATE AND SENNOSIDES

Shrinivas S. and Revanasiddappa M.*

ABSTRACT

A gradient, reversed phase, high pressure liquid chromatography method was developed for simultaneous assay determination of methylparaben, propylparaben, potassium sorbate and sennosides in a single analysis with good resolution between the components. The chromatographic separation is carried out in a reverse phase mode with ultraviolet (UV) detection at 258nm, Column thermo hypersil BDS C18, 250mm × 4.0mm, 5μm using a gradient program run for 40 minutes. Flow rate is 0.7mL/min with ambient column temperature and injection volume is 10.0 μL. The mobile phase consists of 0.5M ammonium acetate buffer adjusted pH to 4.5 units with glacial acetic acid and acetonitrile. The method successfully separates the three preservatives (methylparaben, propylparaben and potassium sorbate) and the active ingredient (sennosides). The developed method was validated according to the international conference on harmonization (ICH) guidelines regarding: Precision, specificity by degradation, linearity, accuracy, range and robustness. The proposed method shown good linearity (correlation coefficient and regression coefficient were not less than 0.999 and 0.998) in the range of 50 to 150% of working concentration of sennosides and 20 to 200% of working concentration of preservatives. The recovery for sennosides at 50, 100 and 150% of working concentration level was within 98 to 102%. The recovery for preservatives at 20, 50, 100, 150 and 200% of working concentration level were within the acceptance criteria. The range of the method is concluded that developed method is from 50 to 150% target concentration of sennosides and 20 to 200% target concentration of methylparaben, propylparaben and potassium sorbate. The validated method is stability indicative, highly selective, simple, accurate, cost effective, and it is applicable for routine quality-control analysis of drug substances as well as drug product in pharmaceutical industries.

Keywords: Methylparaben; Propylparaben; Potassium sorbate: Sennosides: HPLC.


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