A high-pressure liquid chromatographic method for the determination of N-acetyl-p-aminophenol (acetaminophen) in serum or plasma using a direct injection technique.

Abstract

N-Acetyl-p-aminophenol (acetaminophen) is becoming more prevalent as an intoxicant in accidental or intentional overdose, therefore, a direct injection ultra-micro high-pressure liquid chromatographic (HPLC) method has been developed for its quantitation. The HPLC analysis was performed using a Model 110 Solvent Metering Pump equipped with a Model 110-19 Pressure Filter (Altex Scientific, Berkeley, CA), a Model 7120 Rheodyne Injector (Rheodyne, Berkeley, CA) or a Model U6K Injector (Waters Associates, Milford, MA) a Model 440 Absorbance Detector (Water's Associates), and a Model 3380A Recorder Integrator (Hewlett Packard, Avondale, PA). A commercially prepared muBonapak C18 Column (Water's Associates) was used. Acetaminophen was eluted with a mixture of 0.01 mol/L aqueous sodium acetate, pH 4.0: acetonitrile (93:7) and the absorbance detector was operated wih a 254 nm filter. The method, which requires only 2 microL of serum or plasma for analysis, offers several distinct advantages to the analyst. No pre- or post-column extraction or other manipulation of the specimen is required to obtain a quantitative result. Rapid processing of the specimen is possible because both acetaminophen and the internal standard are eluted in less than 10 minutes. The small sample (2 microL) is ideal for use with pediatric patients.