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Russian deer velvet antlers were divided into three parts and subjected to a extraction process using hot water at 100, 110,and 120oC or an extraction with 70% ethanol. Each extract was analyzed for its biochemical components, including uronic acid, sulfated-glycosaminoglycans (sulfated-GAGs), and sialic acid, and the antioxidant and anti-acetylcholinesterase activities were investigated. Different levels of uronic acid and sulfated-GAGs were observed in the extracts according to the water temperature used for the extraction, and contents decreased with increasing extraction temperature. The upper layer of each extract showed high amounts of uronic acid and sulfated-GAGs, followed by the middle and base layers. Ethanol extraction was more effective for recovering uronic acid than sulfated-GAGs. Sialic acid content was the highest in the 110℃ extracts but was not observed in the ethanol extracts. Velvet antler extracts showed strong antioxidant activities against DPPH and hydrogen peroxide as well as strong reducing power in a dose-dependent manner. However, the antioxidant activities were different in each layer and according to the extraction method. Additionally, velvet antler extracts exhibited inhibitory activity against acetylcholinesterase, which is associated with Alzheimer’s disease, in a dose-dependent manner. These results suggest that velvet antler extracts are useful as a functional food ingredient and/or a pharmaceutical.