A robust method for quantifying 42 phenolic compounds by RP-HPLC/DAD: Columns performance and characterization of Brazilian Citrus peels

Reliable analytical methods are the basis for the elucidation of phenolic compounds in foods. This study aimed to optimize and validate a method for determining 42 phenolics using reverse-phase (RP) high-performance liquid chromatography (HPLC) coupled to diode-array-detector-DAD. The performance of two RP columns was evaluated. The 150x4.6 mm 3-μm column showed superior separation quality, whereas 35 of the 42 phenolics showed a separation resolution ≥1.5. The method's linearity, precision (coefficient variation< 3.09%), recovery (87.5-103.2%), specificity, limits of detection (0.04-0.25 mg/L), and quantification (0.06-0.25 mg/L) had acceptable ranges. Thirty phenolics were quantified in Citrus peels, mainly flavanones, flavanols, flavonols, and phenolic acids, highlighting the high values of hesperidin (535-35070 mg/kg) and naringin (26-36466 mg/kg). Lemon peels named 'Lisboa,' 'Thaiti,' 'Thaiti-2000', and 'Thaiti-2001' presented the main phenolics associated with antioxidant capacity. The presented method was robust for determining 42 phenolic compounds, offering a new approach for bioactive compound quantification in food matrices.