3,5-Di-O-methyl-D-glucose has been synthesized by methylation of 1,2-O-isopropylidene-6-O-trityl-D-glucofuranose and removal of the blocking groups by acid hydrolysis. The di-O-methyl glucose furnished a crystalline osazone without loss of a methoxyl group and yielded a monobasic and dibasic acid when oxidized by bromine and nitric acid respectively. Both acids formed gamma lactones and crystalline amides by which the parent compound is characterized. The foregoing reactions proved the structure of 3,5-di-O-methyl-D-glucose predicted by its method of synthesis.