TITLE:
Identification, Isolation and Characterization of Unknown Acid Degradation Product of Nevirapine
AUTHORS:
Vasudev Pottabathini, Vijayacharan Gugulothu, Muralidharan Kaliyaperumal, Satyanarayana Battu
KEYWORDS:
Nevirapine, Method Development, Validation, Forced Degradation, Unknown Impurity, Isolation, Characterization, NMR, HRMS
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.7 No.9,
September
28,
2016
ABSTRACT: A Novel stability indicating RP-UPLC chromatographic method was developed
for analysis of Nevirapine in pharmaceutical formulations. The developed
RP-UPLC method is superior in technology to conventional RP-HPLC with respect
to speed, resolution, solvent consumption and cost of analysis. Nevirapine was
subjected to the stress conditions like acid, base, thermal, oxidative and
photolytic degradation. Nevirapine was found to degrade significantly in acid
and thermal degradation. In acid degradation relative retention time with 0.42
is found as unknown impurity. New impurity was identified, isolated using mass
based auto purification system and characterized by 1H NMR (1D
and 2D) and HRMS experiments. Isolated impurity was showing
molecular weight of 244.10, molecular formula C12H12N4O2 and its name as 2-(3-Amino-4-methylpyridin-2-ylamino)nicotinic
acid. The calibration graph was linear and the method showed less
deviation in accuracy results. The test solution was found to be stable for 20
days when stored in the refrigerator between 2°C to 8°C. The developed RP-UPLC method was validated and
meets the requirements delineated by the International Conference on
Harmonization (ICH) guidelines. The intra-day and inter-day variation was less
than 1%. The method was reproducible and selective for the estimation of Nevirapine. Because the method could
effectively separate the drug from its degradation products, it can be employed
as a stability-indicating method.