Forced degradation of fingolimod: effect of co-solvent and characterization of degradation products by UHPLC-Q-TOF-MS/MS and 1H NMR.
Fingolimod (FGL), an immunomodulator drug for treating multiple sclerosis, was subjected to hydrolysis (acidic, alkaline and neutral), oxidation, photolysis and thermal stress, as per International Conference on Harmonization specified conditions. The drug showed extensive degradation under base hydrolysis, however, it was stable under all other conditions. A total of three degradation products (DPs) were observed. The chromatographic separation of the drug and its degradation products was achieved on a Fortis C18 (100×2.1mm, 1.7μm) column with a mobile phase composed of 0.1% formic acid (Solvent A) and acetonitrile (Solvent B) in gradient mode. All the DPs were identified and characterized by liquid chromatography-quadrupole time of flight-mass spectrometry (LC-Q-TOF-MS) in combination with accurate mass measurements. The major DP was isolated and characterized by Nuclear Magnetic resonance spectroscopy. This is a typical case of degradation where acetonitrile used as co-solvent in stress studies, reacts with FGL in base hydrolytic conditions to produce acetylated DPs. Hence, it can be suggested that acetonitrile is not preferable as a co-solvent for stress degradation of FGL. The developed UHPLC method was validated as per ICH guidelines.
Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research (NIPER), Balanagar, Hyderabad, 500037, Telangana, India.
Prinesh N
PN
Patel
Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research (NIPER), Balanagar, Hyderabad, 500037, Telangana, India.
Pradipbhai D
PD
Kalariya
Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research (NIPER), Balanagar, Hyderabad, 500037, Telangana, India. Electronic address: gana@niperhyd.ac.in.
S
S
Gananadhamu
Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research (NIPER), Balanagar, Hyderabad, 500037, Telangana, India; National Centre for Mass Spectrometry, CSIR-Indian Institute of Chemical Technology, Hyderabad, 500007, Telangana, India. Electronic address: srini@iict.res.in.
R
R
Srinivas
D000097:Acetonitriles / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D000068876:Fingolimod Hydrochloride / Q000737:chemistry*; D005561:Formates / Q000737:chemistry; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007155:Immunologic Factors / Q000737:chemistry*; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010782:Photolysis; D066244:Proton Magnetic Resonance Spectroscopy; D015203:Reproducibility of Results; D012997:Solvents / Q000737:chemistry*; D053719:Tandem Mass Spectrometry; D013678:Technology, Pharmaceutical / Q000379:methods*
D000097:Acetonitriles; D005561:Formates; D007155:Immunologic Factors; D012997:Solvents; C030544:formic acid; D000068876:Fingolimod Hydrochloride; C032159:acetonitrile
Fingolimod; Forced degradation; LC–MS/MS; Stress studies; UHPLC stability assay