TITLE:
Identification, Isolation and Structure Confirmation of Forced Degradation Products of Sofosbuvir
AUTHORS:
Vasudev Pottabathini, Vijayacharan Gugulothu, Muralidharan Kaliyaperumal, Satyanarayana Battu
KEYWORDS:
Sofosbuvir, Forced Degradation, Unknown Impurities, Isolation, Characterization
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.7 No.11,
November
10,
2016
ABSTRACT: A new stability indicating reverse phase chromatographic method was
developed for the analysis of Heptisis C Drug Sofosbuvir. The developed UPLC
method was superior in technology to conventional RP-HPLC with respect to
resolution, speed, solvent consumption and analysis cost. Sofosbuvir was subjected to the thermal,
hydrolytic, oxidative, and photolytic degradation, according to ICH guidelines.
The drug depicted degradation in acidic, basic and oxidative conditions and it
was stable to other stress conditions (thermal and photolytic). Identified
degradation products were isolated using mass supported auto purification
system and characterized by NMR techniques (1H NMR, D2O
Exchange, 13C NMR, 31P NMR, 19F NMR, HSQC and
HMBC) and HRMS experiments. Isolated acid degradation impurity was showing
molecular weight of 416.08, molecular formula C16H18FN2O8P
and its name as (R)-((2R, 3R, 4R, 5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methyl
phenyl hydrogen phosphate. Isolated base degradation impurity-A was showing
molecular weight of 453.13, molecular formula C16H25FN3O9P
and its name as (S)-isopropyl 2-((R)-(((2R, 3R, 4R, 5R)-5-(2, 4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(hydroxy)phosphorylamino)propanoate.
Isolated base degradation impurity-B was showing molecular weight of 411.08,
molecular formula C13H19FN3O9 P and its name as
(S)-2-((R)-(((2R, 3R, 4R, 5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methox-y)(hydroxy)phos-phorylamino)propanoic acid.
In oxidative degradation study, degradation was observed very less and with
obtained quantity (less than 2 mg) recorded 1H NMR and HRMS
analysis. From the spectral data degradation product was showing molecular
weight of 527.15, molecular formula C22H27FN3O9P
and its name as (S)-isopropyl 2-((S)-(((2R, 4S,
5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-4-methyl-3-oxotetrahydrofuran-2-yl)methoxy)(phenoxy)phosphorylamino)propanoate.
The developed chromatographic method was validated as per ICH guidelines and
proved suitable for the stability testing and quality control of the drug
Sofosbuvir