Assay of ibuprofen in human plasma by rapid and sensitive reversed-phase high-performance liquid chromatography:application to a single dose pharmacokinetic study.

A rapid and sensitive reversed-phase high-performance liquid chromatography (HPLC) method is developed to determine the concentration of ibuprofen in human plasma. Ibuprofen is isolated from plasma by adding 0.50 mL of acetonitrile to 1.0 mL of plasma. The endogenous substances precipitated by acetonitrile are separated by centrifugation. The supernatant is saturated with ammonium sulfate to salt-out the acetonitrile. The salted-out acetonitrile is injected directly into the HPLC system. A 150-mm x 4.6-mm column packed with 3-microns reversed-phase octadecylsilane particles (C18) is used for the method finally developed. The mobile phase is a 1:1 ratio of acetonitrile-phosphoric acid (pH 2.2). Ibuprofen is monitored with a UV-visible detector at 220 nm and 0.10-0.002 absorbance units full scale (A.U.F.S.). The mean percent of relative standard deviations for within-day and between-day analyses are less than 3. The limit of detection for ibuprofen (in human plasma) is 25 ng/mL for a 100-microL injection volume. Quantitation of ibuprofen in human plasma at 100 ng/mL can be achieved with a relative standard deviation of less than 5%. The completion time of assay is less than 20 minutes.