Validation of a methodology for the caffeine quantification by liquid chromatography (UHPLC/DAD)
DOI:
https://doi.org/10.56926/repia.v3i1.56Keywords:
Amazon, coffee, caffeine, methodology, UHPLC-DADAbstract
The determination of caffeine in eight coffee samples from different cities of the Peruvian Amazon, Colombia, and Ecuador was carried out developing a methodology for the routine analysis of caffeine content in different samples by reversed-phase high-performance liquid chromatography. This study used a mobile phase of water: methanol (75:25, v/v), at a flow rate of 0.3 mL min-1 and a temperature of 45 °C. Caffeine was isocratically separated at 3.8 min. The caffeine concentrations in the samples were determined by a calibration curve with external standards, which were found to be in the range of 9.74 to 11.12 mg g-1 in roasted ground coffee, at 11.08 mg/g in coffee husk and from 9.17 to 26.59 mg/g in freeze-dried soluble coffee. 68.34, 93.98, and 78.0 % were recovered for the 50, 100, and 150 μg mL-1 fortifications, respectively. These results indicate that the quantification methodology was validated through a simple and rapid separation method. The study made it possible to obtain caffeine concentrations in coffee samples from the region and to suggest average consumption without exceeding healthy limits.
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