ABSTRACT:
In this research, an accurate, efficient and reproducible isocratic reversed-phase high-performance liquid chromatographic (RP-HPLC) method has been developed and validated for the determination of aspirin (ASP), caffeine (CAF), and Paracetamol (PCM). A mixture of drugs as standard material and in formulation tablets were separated using an phenomenex C18 column, (L, 15cm, I.D, 0.46cm, and Size of particle, 5µm), with the shimadzu RP – HPLC, model LC–20 - A, Japan. The eluent phase was optimized through the design, experiment. Elution was done by an eluent phase composed of water (H2O), methanol (MOH) and acetonitrile (ACN) mixture have a ratio of (60: 20: 20, V/V), with adjusted pH of 4.0 with acetic acid, have a pumped flow rate of 0.5mL/min. The drugs, separation were performed at 254nm using a UV-VIS - detector for 8 min. The time of elution for the drugs was recorded at (3.980, 4.366, and 6.894 min) for PCM, CAF and ASP respectively. The validation of the method was conducted in a range of concentration (1 - 100µg/mL) for PCM, CAF and ASP. The method was found to be robust, specified and resisting against the small experimental changes in the value of the flow rate, pH, and wavelength. The LOD were found to be 0.01µg/mL, 0.03µg/mL and 0.05µg/mL for PCM, CAF and ASP respectively. The values of LOQ were 0.033µg/mL, 0.099µg/mL and 0.165µg/mL for PCM, CAF and ASP respectively.