Manihot esculenta roots are considered a staple food for many populations with risk of vitamin A
deficiency. In cassava, β-carotene is the majoritarian carotenoid with pro-vitamin A activity. As a strategy to reduce
the prevalence of deficiency of this vitamin, cassava genotypes with high pro-vitamin A activity have been identified.
The aim of this study was to initiate the metabolomics characterization of a cassava genebank, focusing on the carotenoid
composition of ten cassava genotypes cultivated in southern Brazil by UV-visible scanning spectrophotometry (UV-Vis) and
reverse phase-high performance liquid chromatography (RP-HPLC). The data set was used for the construction of a
descriptive model by chemometric analysis. The results clearly show three genotype groups according to their similarities.
The geno-types from yellow roots were clustered by the higher concentrations of cis-β-carotene and lutein. Inversely,
the genotypes from cream roots were grouped precisely due to their lower concentrations of these pigments. Samples of
red roots rich in lycopene showed to be discrepant among the studied genotypes. The analytical approach (UV-Vis, HPLC,
and chemometrics) herein used showed to be efficient for better under-standing the chemodiversity of cassava genotypes,
allowing to classify them accord-ing to important features for human health and nutrition.