Optimization of Methodology for Simultaneous Quantification of Trigonelline, 5-Caffeoylquinic Acid, and Caffeine in Green and Roasted Coffee Extracts by HPLC

The main bioactive compounds in coffee beans affecting beverage quality include trigonelline, 5-caffeoylquinic acid (5-CQA), and caffeine. Despite the use of various analytical techniques, there is a need for a faster and more accessible alternative to the costly and complex methods used in routine coffee composition analysis. This study aimed to optimize and validate a high-performance liquid chromatography with ultraviolet detection (HPLC-UV-DAD) method for the simultaneous quantification of these three bioactive compounds in five <i>Coffea arabica</i> cultivars (Yellow Catuai, Guesha, Arara, Red Catuai, and Laurina). The evaluated parameters included linearity, accuracy, robustness, precision, limit of quantification, and detection. The proposed method met the criteria of the analytical validation figures of merit, being considered to be fast and effective for determining these compounds. In addition, the study of the concentration of these compounds in the different coffee cultivars showed significant differences, with the Laurina cultivar having the lowest concentration of caffeine (7.9 mg g^-1). Between raw and roasted beans, there were significant variations in the degradation rates of 5-CQA and trigonelline, with the highest for 5-CQA in the Arara cultivar (62.59%) and the highest for trigonelline (28.76%) in the Laurina cultivar. The speed and simplicity of the method can be used to investigate the contribution of these bioactives to the sensory characteristics of the drink and their possible health benefits.