The synthesis of 2-acetamido-2-deoxy-D-glucose 4-phosphate is described. A suitably protected derivative (benzyl 2-acetamido-3-O-acetyl-6-O-benzoyl-2-deoxy-β-D-glucopyranoside) was phosphorylated with o-phenylene phosphochloridate. The removal of the o-hydroxyphenyl group was accomplished by using 2% bromine in buffered aqueous-ethanol, and the ester groups were removed by mild alkaline treatment. Removal of the benzyl glycosidic group by hydrogenation yielded 2-acetamido-2-deoxy-D-glucose 4-phosphate, which was obtained in a crystalline form as its monoammonium salt monohydrate.