Bee pollen (BP) has been increasingly studied because it contains a wide variety of bioactive compounds, including vitamins. Brazilian botanical diversity, together with the potential of local beekeeping production, makes BP's capacity as a food source of bioactive compounds major focus for research. In this scenario, the objective of this study was to screen and evaluate the chemical composition, including antioxidant vitamins, of BP samples from southern Brazil, and to correlate them with their botanical origin. Analyses of nutritional composition were performed to compare them with the quality parameters established by Brazilian and international regulations. Additionally, individual sugars and vitamins (C, E and pro-vitamin A) were quantified and microscopic analysis for taxon classification was performed to correlate with vitamins and nutritional composition. The results of the chemical analysis showed that the samples were in accordance with the relevant regulations. The composition of vitamins and pollen types varied among the samples. Some BP could be classified as a source of a particular vitamin in a standard dose (25 g). Lipid and protein content from Rio Grande do Sul presented higher mean values (p b 0.05) compared with the other two states. Some correlations between chemical composition and botanical taxon were observed. Principal component analysis showed that the samples from the states of Rio Grande do Sul and Paraná presented similarities in terms of composition for each location. HCA and PLSDA were not able to classify the samples based on the chemical markers used. The analysis of vitamins confirmed that BP from this region can be a good source of antioxidant vitamins and that it can provide important nutritional information to food researchers and bioactive compounds for consumers.