Simultaneous Analysis of Major Anthocyanins in Aronia-Containing Food Supplements by HPLC: A Greenness and Whiteness Approach

BACKGROUND: Aronia melanocarpa fruit is a valuable component of a healthy diet due to its rich antioxidant activity and high content of bioactive compounds. Recently, many food products and food supplements have emerged due to the increasing interest in aronia. OBJECTIVE: In this study, a liquid chromatography method for the simultaneous quantification of the anthocyanins cyanidin-3-galactoside, cyanidin-3-arabinoside, and cyanidin-3-glucoside in aronia was developed. METHODS: In this HPLC system, the simultaneous analysis of the three active substances was performed on a Phenomenex, Gemini C18, (150 x 4.6 mm, 5 µm) column at 30 °C. As a mobile phase, 1% glacial acetic acid: ACN: phosphoric acid (510:485:5) (h: h: h) and 0.5% phosphoric acid solutions were used, and a gradient system was applied at a flow rate of 0.8 mL/min. RESULTS: The developed method was validated according to ICH Q2 (R1) guidelines. Cyanidin-3-galactoside and cyanidin-3-arabinoside were found linearly in the concentration ranges of 2.5-12.5 µg/mL, and cyanidin-3-glucoside was found in the range of 0.25-4.0 µg/mL (r > 0.99). CONCLUSION: The developed and validated method was successfully applied to the analysis of samples containing aronia. Moreover, the proposed chromatographic method was assessed in terms of greenness and whiteness using the Analytical Greenness software and the White Analytical Chemistry model. HIGHLIGHTS: A reliable HPLC method was developed for simultaneous quantification of three major anthocyanins in Aronia melanocarpa. The validated method was successfully applied to real aronia-containing samples.