Simultaneous Determination of Acrylamide, Caffeic Acid, and Caffeine in Commercial Coffee Through a Rapid and Validated HPLC Method

Abstract

Background: A simple, fast, precise, accurate, and responsive high-performance liquid chromatography method was developed and validated for the simultaneous quantification of caffeic acid, caffeine, and acrylamide in coffee beverages.

Aims: The chromatographic separation was achieved using a reversed-phase C-18 column (150 x 4.6 mm, 5 µm) with an isocratic mobile phase consisting of methanol: water (40:60, v/v) at a flow rate of 1.5 ml/min. Detection was performed at a wavelength of 210 nm, and an injection volume of 20 μl was used.

Patients and Methods: The method was validated following the guidelines of the International Council for Harmonization (ICH). Specificity, linearity, accuracy, precision, and detection and quantitation limits were assessed. The method demonstrated good linearity for caffeic acid, caffeine, and acrylamide over a wide concentration range.

Results: The percentage recoveries were found to be between 98.19% - 100.77% and 98.59% - 100.75% for the analytes. The percentage of relative standard deviation (RSD) was below 2% for intra-day and 2 % for inter-day variability. Limits of detection were determined to be 0.002 mg/mL for caffeic acid, 0.026 mg/mL for caffeine, and limits of quantitation were 0.006 mg/mL for caffeic acid and 0.086 mg/mL for caffeine.

Conclusions: This method offers a simple, rapid, and highly accurate means of simultaneously assaying caffeic acid, caffeine, and acrylamide in various coffee beverages, making it suitable for routine quality control and safety assessment in the coffee industry.