Three impurities of plerixafor were synthesized with the purity more than 99%,which named 1,11-bis[4-[(1,4,8,11-tetraazacyclotetradecan-1-yl)methyl]benzyl]-1,4,8,11-tetraazacyclotetradecane hydrochloride(related substance A hydrochloride),1,4-bis[4-[(1,4,8,11-tetraazacyclotetradecan-1-yl)methyl]benzyl]-1,4,8,11-tetraazacyclotetradecane hydrochloride(related substance B hydrochloride)and 1,8-bis[4-[(1,4,8,11-tetraazacyclotetradecan-1-yl)methyl]benzyl]-1,4,8,11-tetraazacyclotetradecane hydrochloride(related substance C hydrochloride),and their chemical structure were confirmed by 1H NMR and ESI-MS.The synthesis routes of the related substances A hydrochloride and B hydrochloride had been reported at the first time.They can be used as the references for the quality study of plerixafor.