title: Development of a stability-indicating HPLC method of etifoxine with characterization of degradation products by LC-MS/TOF, 1H and 13C NMR.
abstract: This paper describes a new LC-MS/TOF method for the degradation products determination when Etifoxine (ETI) is submitted to different stress conditions. Chromatography is performed by using Kromasil C18 column (250mm×4.6mm, 5μm particle size). The selected mobile phase consists of formate buffer 0.02M, pH 3 and methanol (70/30, v/v). ETI is submitted to oxidative, acidic, basic, hydrolytic, thermal and UV light degradations. Detection is made at 254nm by photodiode array detector and mass spectrometry. A number of degradation products (DPs) called DPA, DPB, DPC and DPD are found depending on the stress; DPA with heat, DPA and DPB in acidic media or under UV-light; DPA, DPB and DPC under basic stress; DPA, DPB, DPC and DPD with oxidation. LC-MS/TOF is used to characterize the four DPs of ETI resulting from different stress conditions. (1)H and (13)C NMR are used to confirm the DP structures. The ETI fragmentation pathway is proposed. The method is validated with reference to International Conference on Harmonization guidelines and ETI are selectively determined in presence of its DPs, demonstrating its stability-indicating nature. Finally, for the validation step, specificity, linearity, accuracy and precision are determined for ETI and its DPs.
authors.affiliation: Laboratoire de chromatographie, Faculté de chimie, USTHB, BP 32, El-Alia, Bab-Ezzouar, 16133 Alger, Algeria.
authors.forename: Nadia
authors.initials: N
authors.lastname: Djabrouhou
authors.affiliation: Laboratoire de chromatographie, Faculté de chimie, USTHB, BP 32, El-Alia, Bab-Ezzouar, 16133 Alger, Algeria. Electronic address: hguermouche@gmail.com.
authors.forename: Moulay-Hassane
authors.initials: MH
authors.lastname: Guermouche
mesh_terms: D014151:Anti-Anxiety Agents / Q000737:chemistry* / Q000528:radiation effects; D000927:Anticonvulsants / Q000737:chemistry* / Q000528:radiation effects; D002138:Calibration; D002214:Capsules; D066241:Carbon-13 Magnetic Resonance Spectroscopy / Q000592:standards; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D004355:Drug Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D057230:Limit of Detection; D015394:Molecular Structure; D010078:Oxazines / Q000737:chemistry* / Q000528:radiation effects; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010782:Photolysis; D066244:Proton Magnetic Resonance Spectroscopy / Q000592:standards; D012015:Reference Standards; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000592:standards; D013056:Spectrophotometry, Ultraviolet; D013997:Time Factors; D014466:Ultraviolet Rays
chemical_list: D014151:Anti-Anxiety Agents; D000927:Anticonvulsants; D002214:Capsules; D010078:Oxazines; C002125:etifoxine
keywords: (1)H and (13)C NMR; Etifoxine; HPLC; LC-MS/TOF; Stress degradation; Validation