Hydrolytic cleavage of the non-terminal alpha-1,4-glycosidic bonds in alpha-, beta-, and gamma-cyclodextrins and the anomeric-terminal one in D-maltose was investigated to examine how the cleavage rate for alpha-, beta-, and gamma-cyclodextrins is slower than that for D-maltose. Effects of water and temperature were studied by applying in situ C-13 NMR spectroscopy and using a dimethyl sulfoxide (DMSO) water mixture over a wide range of water mole fraction, x(w) = 0.004-1, at temperatures of 120-180 degrees C. The cleavage rate constant for the non-anomeric glycosidic bond was smaller by a factor of 6-10 than that of the anomeric-terminal one. The glycosidic-bond cleavage is significantly accelerated through the keto-enol tautomerization of the anomeric-terminal D-glucose unit into the D-fructose one. The smaller the size of the cyclodextrin, the easier the bond cleavage due to the ring strain. The remarkable enhancement in the cleavage rate with decreasing water content was observed for the cyclodextrins and D-maltose as well as D-cellobiose. This shows the important effect of the solitary water whose hydrogen bonding to other water molecules is prohibited by the presence of the organic dipolar aprotic solvent, DMSO, and which has more naked partial charges and higher reactivity. A high 5-hydroxymethyl-2-furaldehyde (5-HMF) yield of 64% was attained in a non-catalytic conversion by tuning the water content to x(w) = 0.30, at which the undesired polymerization by-paths can be most effectively suppressed. This study provides a step toward designing a new optimal, earth-benign generation process of 5-HMF starting from biomass.