An HPLC method with electrochemical detection in the oxidation mode is developed for the determination of caffeine, theophylline, theobromine, and adenine. The method includes a separation step on RP-C8 column using acetonitrile and 0.05M KH2PO4 buffer solution in a ratio of 10:90% (v/v) as a mobile phase. Electrochemical detection was conducted on a Glassy Carbon Electrode (GCE) and on a self-made Iodine-coated platinum (Pt-I) electrode. The optimum working potential for GCE used with the Ag/AgCl (3M KCl) as reference electrode was found to be at +1.45 V, whereas that for the Pt-I electrode was found to be at +1.55 V. The Pt-I electrode shows equal or better sensitivity compared to GCE. The developed method shows a good applicability (sensitivity, selectivity and reproducibility) for real samples of coffee and tea.
Mahmoud A. Alawi , Safaa Al-Shawa
HPLC/ELCD, Caffeine, Theophylline, Theobromine, Adenine.