Determination of Theobromine, Theophylline, and Caffeine in Various Food Products Using Derivative UV-Spectrophotometric Techniques and High-Performance Liquid Chromatography

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Abstract
Accurate, sensitive and precise derivative ultraviolet Spectrophotometric and high-performance liquid chromatographic (HPLC) methods are described for the simultaneous determination of three natural methyl xanthines (theobromine, theophylline and caffeine) in various food products. The derivative UV-spectro-photometry, with zero crrosing technique of measurement was selected for determination of theobromine and theophylline. First-derivative procedure (D1) was adopted for theobromine determination in the presence of theophylline and caffeine; and second-derivative procedure (D2) was adopted for theophylline determination in the presence of theobromine and caffeine. Caffeine was determined by compensation method using the zero order spectrophotometry. The second method was based on HPLC on a reversed phase column, using a mobile phase of 0.01 M sodium acetate solution: acetonitrile (85:15%) at PH 4.0 with detection at 272 nm. Both methods show good linearity, precision and reproducibility. The developed methods proved to be applicable for the determination of methyl xanthines in various food products. The validity of the proposed methods were tested by analyzing laboratory-prepared mixtures containing the three components in various proportions and by analyzing several food products spiked with a known concentration of the three components.