Simultaneous analysis of phenylephrine hydrochloride, pheniramine maleate, ascorbic acid, paracetamol and related compounds by chromatography

In this thesis, an Ultra Performance Liquid Chromatography (UPLC) method was developed and validated to simultaneous determination of phenylephrine hydrochloride, pheniramine maleate, paracetamol, ascorbic acid, and their impurities in a powder sachet pharmaceutical preparation. Developed ultra performance liquid chromatography method is based on using of reversed phase C18 column. The mobile phase consist of methanol and 10 mM potassium dihydrogen phosphate buffer solution (pH:3.0) by applying a gradient program with UV detector with the flow rate of 0.3 mL/min. The detection wavelength was chosen as 270 nm. Since ascorbic acid impurities could not be detected with this developed method, a High Performance Liquid Chromatography (HPLC) assay and impurity determination method was developed and validated for the analysis of these substances. In this developed method, methanol:10 mM potassium dihydrogen phosphate buffer solution (pH:3.0) (50:50, h:h) isocratically as the mobile phase, at a flow rate of 0.5 mL/min, at a wavelength of 210 nm were applied. The linear ranges are 10-30 µg /mL for phenylephrine hydrochloride, 20-60 µg /mL for pheniramine maleate, 500-1500 µg /mL for paracetamol, 50-150 µg /mL for ascorbic acid for assay analysis and 0,05-0,3 µg /mL, 0,08-0,6 µg /mL, 2,4-15,0 µg /mL, 0,16-1,5 µg /mL respectively for impurity analysis. The concentration ranges are for phenylephrine hydrochloride impurities C and E 0,003-0,3 µg /mL and 0,002-3,0 µg /mL, for impurities pheniramine maleate A and B 0,11-0,6 µg /mL and 0,03-0,6 µg /mL, for impurities paracetamol J, F, K, 3,5-15,0 µg /mL, 0,9-15,0 µg /mL and 1,7-15,0 µg /mL and for ascorbic acid impurities C and D 0,08-1,5 µg /mL and 0,17-1,5 µg /mL. The detection and quantification limits are 0,008 and 0,047 μg/mL for phenylephrine hydrochloride, 0,014 and 0,042 μg/mL for pheniramine maleate, 0,012 and 0,024 μg/mL for paracetamol, 0,004 and 0,013μg/mL for ascorbic acid. Accuracy studies were found to be between 50-150% for assay analysis and LOQ-150% concentrations for impurity analysis. The developed method is easy, sensitive, fast, reproducible and can be used safely in simultaneous analysis of active substances and their impurities in pharmaceutical preparations.