Simultaneous determination of epinephrine, noradrenaline and dopamine in human serum samples by high performance liquid chromatography with chemiluminescence detection

A simple, rapid and accurate high performance liquid chromatographic (HPLC) technique coupled with chemiluminescence (CL) detection was developed for the simultaneous determination of epinephrine (E), noradrenaline (NA) and dopamine (DA). It was based on the analyte enhancement effect on the CL reaction between luminol and potassium ferricyanide. The effects of various parameters, such as potassium ferricyanide concentration, luminol concentration, pH value and component of the mobile phase on chromatographic behaviors of the analytes (E, NA and DA) were investigated. The separation was carried out on C-18 column using the mobile phase of 0.01 mol/L potassium hydrogen phthalate solution and methanol (92 : 8, V/V). Under the optimum conditions, E, NA and DA showed good linear relationships in the range of 1 X 10(-8) -5 X 10(-6), 5.0 X 10(-9)-1.0 X 10(-6) and 5.0 X 10(-9)- 1.0 X 10(-6) g/mL respectively. The detection limits for E, NA and DA were 4.0 X 10(-9), 1.0 X 10(-9) and 8.0 X 10(-10) g/mL. The proposed method has been applied successfully to the analysis of E, NA and DA in human serum samples.