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-1 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-1 with the determination coefficient (r2) of
0.9999, also accuracy and precision were found to be <2 (%RSD). The intraday
accuracy for concentrations 62.5 μg·mL-1, 15.625 μg·mL-1,
7.812 μg·mL-1 and 1.953 μg·mL-1 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.