High performance liquid chromatographic analysis of caffeine concentrations in plasma and saliva

A rapid high performance liquid chromatographic (HPLC) method for the analysis of caffeine in plasma and saliva is described. Samples of saliva and plasma were purified using zinc sulphate solution as protein precipitant. The supernatant was injected directly onto the column. The mobile phase consisted of ammonium acetate buffer:acetonitrile:methanol (82:15:3, v/v). Measurements were carried out at 254 nm. Acetanilide was used as the internal standard and analysis was completed in 10 min. No interference from endogenous components or other methylxanthines was observed. The coefficients of variation for within day and between day analysis for both saliva and plasma were less than 7.66%. Samples were collected from 20 volunteers. The correlation coefficient between plasma and saliva caffeine concentrations was found to be 0.98.