Abstract

VALIDATED HPLC METHOD FOR DETERMINITION OF 5-AMINO- 2-METHYL BENZOIC ACID IN CANAGLIFLOZIN DRUG SUBSTANCES

Ritesh Kumar Srivastava* and S.Senthil Kumar

ABSTRACT

A sensitive, cost-effective, reproducible HPLC method was developed and validate for the quantitative determination of genotoxic impurity 5- amino-2-methyl benzoic acid present in canagliflozin drug substance by applying the concept of threshold of toxicological concern, a limit of 5.0μg/g was calculated based on the maximum daily dose of the drug substance. The impurity was separated on X-terra MS C18, 250 mm x 4.6mm, 5μm analytical column with a mobile phase consisting of the 0.1 % Orthophosphoric acid and methanol with the gradient program at a flow rate 1.0mL/min. The effluent was monitored by UV detection at 228 nm with column temperature maintained at 35 °C and the injection volume 30μL. Validation activity was planned and completed based on the ICH guideline. The LOD and LOQ value were found to be 0.52μg/g and 1.51 μg/g and accuracy results were well in the range 97.99 to 103.46 %. The linearity curve showed the correlation coefficient of 0.9996 and method very sensitive.

Keywords: Impurity, HPLC, TTC, Validation, ICH guidelines.