Determination of Ketoprofen in Human Plasma by RP-HPLC

For in vivo pharmacokinetic studies, it is pre-requisite to quantify drug concentrations in plasma. In the present study a RP-HPLC procedure was developed and validated for the assessment of ketoprofen in human plasma. For this purpose mobile phase consisting of methaol:water (70:30) adjusted to pH 3.3 with phosphoric acid was used, and chromatography was carried out on Discovery HS C18 column, 5 μm (25 cm × 4.6 mm). The flow rate was 1 mL·min and quantitative assessment was performed at 260 nm. The retention time was found to be <10 min. A method was found to be accurate and illustrated linearity from 0.2441 to 125 μg·mL with the determination coefficient (r) of 0.9999, also accuracy and precision were found to be <2 (%RSD). The intraday accuracy for concentrations 62.5 μg·mL, 15.625 μg·mL, 7.812 μg·mL and 1.953 μg·mL were found to be 99.747%, 99.475%, 98.457% and 99.824% respectively where as for interday accuracy consecutive values for days 1, 2 and 3 were 99.104%, 99.091%, 98.96% and 99.385% in plasma. All validation parameters were assessed and were found to be within the limits. The proposed method was accurate, specific, quick (retention time < 10 min), selective (showed no interference with excipients), cost effective and a good resolution which gave this method an advantage over the different other reported methods for the estimation of ketoprofen in human plasma.