The purity of maltotriosyl-β-cyclodextrin was confirmed by HPLC and the molecular weight was calculated by ESIMS.The chemical composition and structure of maltotriosyl-β-cyclodextrin were elucidated by FTIR,analyzed by enzymatic hydrolysis,1H NMR,13C NMR and HMBC.The results indicated that the purity of maltotriosyl-β-cyclodextrin was more than 99%,its molecular weight was 1620.5 and its structure was mono-6-O-α-D-maltotriosyl-β-cyclodextrin.