Abstract
STABILITY INDICATING RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF ASCARBIC ACID AND N-ACETYL CYSTENE IN BULK AND PHARMACEUTICAL FORMULATION

Aruna Gundala*, Hari Krishna N. and Jayachandra Reddy P.

ABSTRACT

The present study was designed to develop and validate a simple, sensitive, precise and accurate stability indicating RP-HPLC method for simultaneous estimation of Ascorbic acid and N-Acetylcysteine in bulk and tablet dosage form. The chromatographic separation was achieved on Denali C18 (4.6 x 250mm, 5μm) as stationary phase with a mobile phase of 0.1% OPA: acetonitrile (50:50 v/v) at a flow rate of 1 ml/min and PDA detection at 220 nm. The proposed method was validated for system suitability, specificity, linearity, accuracy, precision, LOD, LOQ and robustness as per ICH guidelines. The retention times of Ascorbic acid and N-Acetylcysteine were found to be 2.587 ± 0.06 and 3.254 ±0.05 min respectively. The calibration curves were linear in the concentration range of 25% to 150% of the working concentration (r2=0.999) for both the drugs in binary mixture. The accuracy was found to be 99.45% and 99.31% for Ascorbic acid and N-Acetylcysteine respectively. The LOD was found to be 0.51 μg/ml and 0.24 μg/ml and LOQ was found to be 1.55 μg/ml and 0.73 μg/ml for Ascorbic acid and NAcetylcysteine respectively. The percentage recoveries for both drugs were in the range of 99-100%. Hence the proposed stability indicating RP-HPLC ethod can be used in routine analysis of tablets containing Ascorbic acid and N-Acetylcysteine.

Keywords: Ascorbic acid, N-Acetylcysteine, RP-HPLC, stability, Method development and Validation.


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