ISOCRATIC LC-UV, LC-MS AND MS/MS STUDIES ON FENOXAZOLINE AND ITS DEGRADATION PRODUCTS
Keywords:
Fenoxazoline hydrochloride, Impurities, Forced degradation, Stability indicating method, TLC-MSAbstract
Objective: To develop and validate a selective stability indicating high performance liquid chromatographic method for fenoxazoline hydrochloride, to carry out its forced degradation and to identify the degradation products using hyphenated techniques like liquid chromatography-mass spectrometry, thin layer chromatography-mass spectrometry and tandem mass spectrometry.
Methods: The optimised chromatographic conditions involved the use of Methanol: 10 mM Ammonium acetate (v/v) with 0.3% triethylamine with pH adjusted to 4 using formic acid (55:45) and 0.5 ml/min flow rate. Fenoxazoline hydrochloride was subjected to hydrolytic, oxidative, photolytic and thermal stress conditions as per International Conference on Harmonisation guidelines. The degradation products were characterized by liquid chromatography-mass spectrometry, tandem mass spectrometry and a novel hyphenated technique, thin layer chromatography-mass spectrometry. Fragmentation pathways and degradation pathways of the drug and the degradation products were postulated.
Results: The drug was found to be highly susceptible to degradation under alkaline conditions. Two degradation products were formed in all the stress conditions with aqueous media. This method was validated as per ICH guidelines and was found to comply with the standard norms. The calibration curve was found to be linear between 0.07-100 μg/ml. Limit of detection and quantitation was 0.02 μg/ml and 0.07 μg/ml respectively.
Conclusion: A simple, isocratic and selective stability-indicating high performance liquid chromatographic method has been developed and validated for the determination of fenoxazoline hydrochloride. The validated method was found to be linear, precise, accurate and robust. The degradation products were identified as 2-(2-isopropylphenoxy)-N-(2-aminoethyl) acetamide and 2-(2-isopropylphenoxy) acetic acid, which are novel impurities for fenoxazoline hydrochloride.
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