The Creation and Validation of a New High-Performance Liquid Chromatography Method for Analyzing Pyridostigmine Bromide and its Degradation Products in a Pharmaceutical Formulation

A novel, rapid, sensitive, and validated high-performance liquid chromatographic (HPLC) technique, with high selectivity for stability indication, has been employed to determine pyridostigmine bromide and its degradation products (3-hydroxy-1-methylpyidin-1-ium bromide and 3,4-dioxo-3,4-dihydropyeidin-1-ium) in a pharmaceutical formulation. Pyridostigmine bromide and its degradation products were identified using combinations of acetonitrile and 0.01M acetate buffer (pH=6)in a ratio of 5:95 (v/v), through flow rates of about 1 ml/min at room temperature, employing a diode array detector. The linear range was determined to be 27-135µg/ml. Our findings indicate that pyridostigmine bromide is unstable in alkalineconditions but more stable in acidic environments and susceptible to oxidation.