Rapid high-performance liquid chromatographic assay of acetaminophen in serum and tissue homogenates

The quantitation of a hepatorenal toxic drug, acetaminophen, in blood and target organ tissues is needed for toxicokinetic and distribution studies. A rapid, sensitive and simple method is described to assay acetaminophen in rat serum and liver or kidney homogenates by reversed-phase high-performance liquid chromatography, using an octadecyl (3 micron particle size) Apex column, a mobile phase consisting of a mixture of distilled water-acetonitrile (86:14) and ultraviolet detection at 245 nm. Short retention times of ca. 3.75 and 6.25 min are observed for acetaminophen and the internal standard (sulfamerazine), respectively. A sensitivity of 50 ng/ml is easily achieved for 100-microliter serum and liver or kidney homogenate samples. The proposed method proved to have satisfactory recovery, precision and accuracy. The preliminary results obtained with human plasma of volunteers and of patients treated with various drugs show that the assay, with a sensitivity of 25 ng/ml, would be of considerable interest in clinical monitoring of acetaminophen.