A rapid and selective cation exchange chromatographic method coupled to integrated pulsed amperometric detection (PAD) has been developed
to quantify biogenic amines in chocolate. The method is based on gradient elution of aqueous methanesulfonic acid with post column addition
of strong base to obtain suitable conditions for amperometric detection. A potential waveform able to keep long time performance of the Au
disposable electrode was set up. Total analysis time is less than 20 min. Concentration levels of dopamine, serotonin, tyramine, histamine and
2-phenylethylamine were measured, after extraction with perchloric acid from 2 g samples previously defatted twice with petroleum ether. The
method was used to determine the analytes in chocolate real matrices and their quantification was made with standard addition method. Only
dopamine, histamine and serotonin were found in the analysed real samples. Repeatabilities of their signals, computed on their amounts in the
real samples, were 5% for all of them. Repeatabilities of tyramine and phenethylamine were relative to standard additions to real samples (close
to 1 mg/l in the extract) and were 7 and 3%, respectively. Detection limits were computed with the 3 s of the baseline noise combined with the
calibration plot regression parameters. They were satisfactorily low for all amines: 3 mg/kg for dopamine, 2 mg/kg for tyramine, 1 mg/kg for
histamine, 2 mg/kg for serotonin, 3 mg/kg for 2-phenylethylamine.