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In this study, we examined the effects of different water activities (Aw: 0.3, 0.5, and 0.8) on the lipid composition and oxidation of wheat flour after 28 days of storage in the dark at 60oC. The lipid content of the flour was 2.7%, and had decreased significantly (p<0.05) at the end of the storage period. Decrease in monoacylglycerol and increase in free fatty acids were observed, however, phosphatidic acid, phosphatidylglycerol, and phosphatidylinositiol were not detected after storage. Phosphatidylehtanolamine was more stable than phosphatidylcholine during the dark storage of flour. The flour lipids consisted of palmitic (18%), stearic (1%), oleic (14%), linoleic (63%), and linolenic (4%) acids, and the relative content of linolenic acid decreased after 28 days of storage. The conjugated dienoic acid content of the flour lipid had increased due to lipid oxidation during dark storage. Hydrolysis of neutral lipids and glycolipids, and lipid oxidation, were higher in the flour stored at Aw 0.8 than in the flour stored at Aw 0.3 or 0.5.