The complete assignment of the 1H and 13C sugar resonances in mono-3,6-anhydro-heptakis(2,3-O-methyl)-hexakis(6-O-methyl)-β-cyclodextrin, an asymmetrically functionalized β-cyclodextrin, was carried out by means of 2D NMR experiments. The TOCSY and the homonuclear multiple relay COSY spectra provided most of the 1H assignments. The multiplicity edited HSQC and a set of F1 selective HSQC-TOCSY and multiple relay HSQC-COSY spectra gave access to most of the 13C chemical shifts. The latter were fully and accurately determined by means of a pair of complementary, highly folded HSQC-TOCSY spectra. The TOCSY-ROESY and ROESY-TOCSY spectra yielded the sequential assignment of the sugar units. A high resolution F1 selective F1 decoupled version of the TOCSY-ROESY experiment was recorded.