The polyphenol content of propolis has received a lot of attention due to the benign biological properties noted in the chemical composition studies. However, there are very limited studies about other chemical components found in trace amounts in nature which contribute to the therapeutic properties of propolis. The present study, therefore, investigated the amino acid and vitamin composition of propolis. Propolis harvested by 60 colonies of Apis mellifera caucasica belonged to local non-migratory beekeepers. The A. m. caucasica is known for its distinctive propolis collecting capability which native to the secluded Ardahan Province of Turkey. Vitamin (Thiamine, Riboflavin) combinations of propolis were determined using the HPLC (High-Performance Liquid Chromatography) fluorescent detector. An amino acid analysis was also performed with the UFLC (Ultra-Fast Liquid Chromatography) system consisting of binary pump and UV/VIS. Our findings record that the vitamin and amino acid content of propolis samples collected from three areas of different altitudes in the same region differed from each other. Vitamin B1 content and Vitamin B2 content ranged between 0.025-0.16 mg/100g, and 0.304-0.777mg/100g, respectively. A maximum amount of amino acid was reported as leucine, while a minimum amount of amino acid was seen as tryptophan in Ardahan propolis. Consequently, the vitamin and amino acid content of propolis, which derived from secondary plant metabolites of resin, varied depending on their geographical altitudes. Those vitamin and amino acids found in the propolis composition are believed to have beneficial therapeutic properties.