Abstract

DEVELOPMENT, VALIDATION AND STABILITY INDICATING OF UV - SPECTROSCOPY METHOD FOR SIMULTANEOUS ESTIMATION OF ASCORBIC ACID AND PIPERINE

Rizwan Ahamad, Nazreen Tabassum, Rama K. P., Shubham Singh, Gurpreet Singh, Mohd. Akhtar, Mohd Aqil, Mohd. Mujeeb*

ABSTRACT

Antioxidants like vitamins, polyphenols, and flavonoids protect the body by neutralizing harmful free radicals from various sources. Ascorbic acid (ASA) and piperine (PP) are well-studied antioxidants, playing key roles in several diseases. Owing to their exciting synergistic potential and combination delivery applications, we developed a simple and rapid UV spectroscopy method based on isosbestic point detection. A linear relationship was identified between absorbance readings and corresponding concentrations ranging from 0-30 μg/ml, with correlation coefficients (R²) of 0.995 for ASA and 0.998 for PP respectively. The detection limits were determined at 4.56μg/ml and 2.89μg/ml, while the quantitation limits were set at 14.12μg/ml and 8.77μg/ml for ascorbic acid and piperine, respectively. The recovery percentages of 50%, 100 % and 150%varied from 49.3% to 51%, 102.4% to 102.8% and 150% to 150.7%for both, accompanied by correspondingly low %RSD values, signifying the analytical method's high accuracy. The percent relative standard deviation of ASA and PP mixture, the intra-day analysis was determined and found to be 1.034% at λ1 and 1.121% at λ2, respectively. In the inter-day study, the % RSD was found to be 1.075% at λ1 and 1.679% atλ2, respectively. From the findings, we conclude that our method is both precise and accurate. We further validated our method through forced degradation studies under acidic, alkaline, oxidative, photolytic, and thermal conditions, confirming the susceptibility of both compounds to degradation. These findings underscore the reliability of our simultaneous UV spectroscopic approach for stability analysis and future pharmaceutical research.

Keywords: UV spectroscopy, Simultaneous estimation, Ascorbic acid, Piperine, Validation.