ABSTRACT
The objective of the current study was to develop a validated, specific and stability-indicating reverse phase liquid chromatographic method for the quantitative determination of acetazolamide and its related substances. The determination was done for an active pharmaceutical ingredient, its pharmaceutical dosage form in the presence of degradation products, and its process-related impurities. The drug was subjected to stress conditions of hydrolysis (acid and base), oxidation, photolysis and thermal degradation as per International Conference on Harmonization (ICH) prescribed stress conditions to show the stability-indicating power of the method. Significant degradation was observed during acid and base hydrolysis, and the major degradant was identified by LC-MS, FTIR and (1)H/(13)C NMR spectral analysis. The chromatographic conditions were optimized using an impurity-spiked solution and the generated samples were used for forced degradation studies. In the developed HPLC method, the resolution between acetazolamide and, its process-related impurities (namely imp-1, imp-2, imp-3, imp-4 and its degradation products) was found to be greater than 2. The chromatographic separation was achieved on a C18, 250mmx4.6mm, 5microm column. The LC method employed a linear gradient elution, and the detection wavelength was set at 254nm. The stress samples were assayed against a qualified reference standard and the mass balance was found to be close to 99.6%. The developed RP-LC method was validated with respect to linearity, accuracy, precision and robustness.
Subject(s)
Acetazolamide/analysis , Carbonic Anhydrase Inhibitors/analysis , Chromatography, High Pressure Liquid , Chromatography, Reverse-Phase , Drug Contamination , Technology, Pharmaceutical/methods , Buffers , Chromatography, High Pressure Liquid/standards , Chromatography, Reverse-Phase/standards , Drug Stability , Guidelines as Topic , Hydrogen-Ion Concentration , Hydrolysis , Magnetic Resonance Spectroscopy , Mass Spectrometry , Oxidation-Reduction , Photolysis , Reference Standards , Reproducibility of Results , Spectroscopy, Fourier Transform Infrared , Technology, Pharmaceutical/standards , TemperatureABSTRACT
An experimental technique based on a modified vibrating orifice aerosol generator has been employed to study unsteady evaporation of linear streams of highly monodisperse binary microdroplets of volatile constituents over short time periods (i.e., <1 ms), such that the droplet composition remains nearly constant. The droplet size and temperature (i.e., refractive index) have been determined with high temporal resolution from the resonances observed in the simultaneous elastic and Raman light scattering spectra obtained by varying the droplet size through modulation of droplet generation frequency. By using this technique we show that thermodynamic parameters of binary systems, such as activity coefficients as well as vapor pressures of the constituents as functions of temperature, can be determined. We have applied the procedure to study unsteady evaporation rates of pure ethanol and methanol droplets as well as binary droplets containing various ratios of ethanol and methanol. We have obtained vapor pressures of ethanol and methanol as functions of temperature as well as activity coefficients of ethanol and methanol as functions of composition, and the results show excellent agreements with the values reported in the literature. The technique presented in this paper is applicable to any binary system containing at least one volatile constituent. Copyright 2000 Academic Press.
