The objective of this work is to study the feasibility of a more objective and rigorous classification of the calices of Hibiscus sabdariffa based on their physicochemical profile. To do so, 19 analyses were carried out on 4 varieties of calices cultivated in Senegal: Vimto, Koor, Thaï and CLT92. Principal component analysis results showed that 15 physicochemical and biochemical parameters could be potentially used to discriminate the varieties of calices. Polyphenolic and anthocyanin contents were anti-correlated to protein content and could be used to differentiate the Vimto/CLT92 and the Koor/Thaï varieties. Within these two clusters, pH and lipid content could discriminate each variety. Finally, factorial discriminant analysis showed that total anthocyanin content, lipid content and chromaticity C* were the 3 parameters enabling the most efficient classification of calices according to variety and led to 100% classification accuracy.