This study aimed to investigate the botanical diversity in bee pollen and amino acid composition of Turkish bee pollen samples collected from different regions. The monofloral bee pollen samples belonged to three different plant families: Asteraceae, Fabaceae, and Ranunculaceae. Additionally, Agean, Central Anatolian, Black Sea, and Marmara Regions have monofloral bee pollen samples with higher than 85% pollen frequency. The free amino acid (FAA) composition and content of the bee pollen samples were analyzed, with the essential amino acids comprising 13.13–18.28% of the total amino acid content. The Central Anatolian region displayed the highest ratio of essential amino acids, and the Aegean region exhibited a more diverse profile of FAA. Principal Component Analysis was performed to evaluate the variation in the data, with the eigenvalues of the two factors explaining 68.58% of the total variability. Moreover, cluster analysis revealed distinct dendrograms based on palynological characteristics and FAA composition.