title: Isolation and structure elucidation of major alkaline degradant of ezetimibe.
abstract: This work presents the isolation and characterization of the alkaline degradant of Ezetimibe. Ezetimibe, a selective inhibitor of intestinal cholesterol absorption, was subjected to alkaline degradation. Ezetimibe was reacted with 0.1M methanolic sodium hydroxide solution for 10min at 80°C to yield alkaline degradant to an extent of 90% of initial amount of the drug taken. This degradant was detected by high performance liquid chromatography (HPLC) at relative retention time (RRT) of 1.48 with respect to Ezetimibe. HPLC method involved an isocratic elution on a Waters Symmetry C(8) 150mm×4.6mm, 5μm column using ammonium acetate buffer (pH 4.5, 50mM) - acetonitrile (50:50, v/v) as the mobile phase at a flow rate of 1.0mL/min and UV detection at 242nm. The degradant was isolated by preparative HPLC. Purity of the isolated solid was found to be more than 99%. Structure of alkaline degradant was confirmed by LC-MS, (1)H and (13)C NMR and IR spectroscopy. On the basis of spectral data, the structure of the degradant was confirmed as 5-(4-fluorophenyl)-2-[(4-fluorophenyl amino)-(4-hydroxyphenyl)methyl]-pent-4-enoic acid. The route for the formation of this degradant is also proposed. Determining the structures of degradation products arouse during stress testing can be useful for preclinical discovery efforts.
authors.affiliation: Department of Pharmaceutical Chemistry, Institute of Pharmacy, Nirma University, Ahmedabad 382481, Gujarat, India. anuradha.gajjar@nirmauni.ac.in
authors.forename: Anuradha K
authors.initials: AK
authors.lastname: Gajjar
authors.forename: Vishal D
authors.initials: VD
authors.lastname: Shah
mesh_terms: D000468:Alkalies / Q000009:adverse effects; D001384:Azetidines / Q000737:chemistry* / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D000069438:Ezetimibe; D005765:Gastrointestinal Agents / Q000737:chemistry* / Q000302:isolation & purification*; D006845:Hydrocarbons, Fluorinated / Q000737:chemistry* / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000037:antagonists & inhibitors*; D049990:Membrane Transport Modulators / Q000737:chemistry* / Q000302:isolation & purification*; D026901:Membrane Transport Proteins; D015394:Molecular Structure; D010421:Pentanoic Acids / Q000737:chemistry* / Q000302:isolation & purification*; D021241:Spectrometry, Mass, Electrospray Ionization; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: C558758:5-(4-fluorophenyl)-2-((4-fluorophenylamino)-(4-hydroxyphenyl)methyl)pent-4-enoic acid; D000468:Alkalies; D001384:Azetidines; D005765:Gastrointestinal Agents; D006845:Hydrocarbons, Fluorinated; D008565:Membrane Proteins; D049990:Membrane Transport Modulators; D026901:Membrane Transport Proteins; C410800:NPC1L1 protein, human; D010421:Pentanoic Acids; D000069438:Ezetimibe