ArticleACS omega2025
A New Segmented Gradient HPLC-DAD and Spectra Index Plots for Measuring Caffeine and Chlorogenic Acid in Dark and Green Coffees.
Article in ACS omega, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
A rapid and sensitive segmented gradient elution high-performance liquid chromatographic methodology with a diode array detector (SGE-HPLC-DAD) has been developed for quantifying important bioactive compounds, chlorogenic acid (ChGA) and caffeine (CAFF), in different coffee samples. A Luna cyano-column (5 μm, 25 × 0.46 cm) conditioned at ambient temperature (22 ± 1 °C) and a gradient elution system consisting of solvents A (1% trifluoroacetic acid in water) and B (acetonitrile) were employed. The elution process used a segmented gradient program, starting with a gradual increase in solvent B from 5% to 8% over the first 4 min. This was followed by a rapid ramp-up from 8% to 100% over 1 min (4-5 min) and then an isocratic elution at 100% B lasting 5-7 min. Thereafter, there was a linear decrease from 100% to 5% B over the next minute 7-8 min and finally an isocratic elution by a mixture of A and B (95:5, v/v) until 11 min. The flow rate was maintained at 1.5 mL/min with effluent detection at 254 nm. The SGE-HPLC-DAD method revealed linear relationships between peak areas and concentrations, with CAFF ranging from 0.4 to 1000 μg/mL and ChGA ranging from 0.6 to 1000 μg/mL. The recovery percentages for CAFF and ChGA were estimated to be between 100.97% and 101.33%, respectively. The relationship between ChGA and CAFF contents in dark and green coffees commercially available in the Egyptian market that were brewed using stainless-steel and glass cookware materials on direct heat was also investigated.
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