HPLC Determination of Caffeine and Theobromine in Coffee, Tea, and Instant Hot Cocoa Mixes

Popular brands of instant and ground coffee, instant and bag tea, and instant hot cocoa mixes were prepared according to manufacturer's directions and analyzed for caffeine and/or theobromine content. After extraction into boiling water, the methylxanthines were separated by high performance liquid chromatography using a reverse phase C₁₈ column and a mobile phase of acetonitrile and water (8:92, v/v). Caffeine content ranged from 32.4–35.0 mg/cup in instant tea, 30.2–67.4 mg/cup in bag tea, 1.0–7.8 mg/cup in instant hot cocoa mixes, 46.7–67.6 mg/cup in instant coffee, and 93.0–163.5 mg/cup in ground coffee. Theobromine concentrations ranged from 1.4–2.3 mg/cup in instant tea, 1.2–4.4 mg/cup in bag tea, and 39.5–79.5 mg/cup in instant hot cocoa mixes.