RP–HPLC Method Validation for Simultaneous Estimation of Paracetamol and Caffeine in Formulating Pharmaceutical Form

Abstract

A novel, simple and accurate, Reversed Phase High Performance Liquid Chromatography (RP-HPLC) method for simultaneous estimation of Paracetamol (PCM) and Caffeine (CAF) in mixture of standard and formulation tablets was validated in this research. The absorbance maximum of drugs using UV- spectroscopy was found at (244.8 and 272.6nm) for PCM and CAF respectively in deionized water: methanol mixture (60:40 V/V) as solvent. This method involves the separation of PCM and CAF on RP - HPLC Shimadzu type LC–20 - A, Japan, and Phenomenex column, C18 (250mm, 4.6mm and 5μm). The elution was done using an eluent phase composed of methanol and water in the ratio of (40:60 V/V with a pH adjusted at 4.0 using acetic acid). A separation was fixed for 10 min at 270nm, using a UV-Vis - detector and 1.0mL/min, flow rate and the drugs were eluted in (3.468 and 5.376 min) for PCM and CAF respectively. The suitable conditions such as the elution phase composition, rate of flow, pH and wavelength were studied. The linearity of the method was in the range of concentration within (0.5 – 25 and 0.1 - 30μg/mL), while, R2 values within (0.9995 and 0.9997), and the means of recovery were found within (99.57 and 100.36) for PCM and CAF respectively. The method was applied for the estimation of gradient active of drugs in different formulating form samples. The method accuracy was validated by the mean of recovery percentages which, were found in acceptable limit.