Ketoprofen (KET) is an active pharmaceutical compound that has pain relieving and antipyretic effects. Its determination in body fluids and environmental waters is important due to widespread use of the compound. In this study, a selective and reliable method has been developed for the determination of ketoprofen in water and artificial serum using molecularly imprinted polymers (MIPs) as a solid phase extraction sorbent prior to HPLC-DAD detection. The MIP was synthesized by copolymerization of methacrylic acid (MAA) and trimethylpropane trimethacrylate (TRIM) in the presence of ketoprofen as the template. For the sake of comparison, nonimprinted polymer (NIP) was also synthesized under the same experimental conditions without the addition of ketoprofen under the same experimental conditions. Critical extraction parameters such as sample pH, shaking time and sorbent amount were optimized and adjusted to 8.0, 24 h, and 10.0 mg, respectively, for a sample volume of 10.0 mL. MIP showed higher selectivity than NIP towards ketoprofen in an artificial matrix containing another pain relieving drug, ibuprofen, and a cardiovascular drug, metoprolol. The proposed method was successfully applied for the detection of ketoprofen in spiked drinking water, tap water, and artificial serum samples, and showed satisfactory results with respective recoveries of 96.8 % (± 0.8), 93.7% (± 0.6), 62.2% (± 0.6), and 69.9% (± 0.6).