Determination of ibuprofen in serum by high-performance liquid chromatography and application to ibuprofen...

A high-performance liquid chromatographic method for quantitation of ibuprofen from serum and application of this method to ibuprofen disposition in the dog is described. The drug was extracted from acidified plasma with dichloromethane. The internal standard used was a methanolic solution of 4-n-butylphenylacetic acid. A muBondapak C18 column was used for analysis; this mobile phase was methanol-water-glacial acetic acid (pH 3.4) (75 : 24 : 1, v/v). A wavelength of 272 nm was used to monitor ibuprofen and the internal standard. Method sensitivity was 0.5 microgram/ml serum using either 0.5 or 1.0 ml of sample, and no interference was found from endogenous compounds or other commonly used anti-inflammatory agents. The coefficients of variation of the method were 4.2% and 6.0% for sample containing 50.0 and 6.25 microgram/ml of ibuprofen, respectively, and the calibration curve was linear for the range of 0.5 to 100 microgram/ml. This method was demonstrated to be suitable for pharmacokinetic and/or biopharmaceutical studies of ibuprofen in man and the dog.