Studying the chromatographic separation of caffeine, theophylline, and theobromine
DOI:
https://doi.org/10.31699/IJCPE.2025.2.15Keywords:
HPLC; Mobile phase; Stationary phase; Retention time; ResolutionAbstract
Theobromine, theophylline, and caffeine retention times in a C18 HPLC column (stationary phase) were investigated as a function of mobile phase flow rate, mobile phase composition, and column temperature. When the mobile phase flow rate increased from 0.1 to 1 mL/min and the methanol concentration increased from 5 to 30%, the retention time and peak width of these three compounds were found to be reduced. While there was a small influence of increasing the mobile phase flow rate on the resolution, decreasing the methanol concentration in the mobile phase considerably reduced the resolution. In addition, the mobile phase flow rate and composition were determined to have a more significant impact than the column temperature. According to the findings, theobromine, theophylline, and caffeine were most effectively separated by liquid chromatography at a flow rate of 0.5 mL/min with a mobile phase methanol concentration of 15%.
Received on 06/04/2025
Received in Revised Form on 10/06/2025
Accepted on 10/06/2025
Published on 30/06/2025
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