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Poly(vinyl alcohol) (PVA) microspheres containing cyclodextrin (CD) were obtained by chemical cross-linking with glutaraldehyde of an acidified mixture solution of PVA and α-, β- or γ-CD. The amount of linked CD in microspheres, estimated by tetrazolium blue method, decreases in the order β- > γ- > α-CD. The dimensions of PVA/γ-CD microspheres are much higher than those of PVA/α- and β-CD. The cross-linking density of microspheres was estimated by the amount of iodine retained by the polymer matrix. The pore size as well as the porous volume of PVA/CD microspheres decrease significantly on increasing the amount of glutaraldehyde, but are enough large to permit the access of drugs to the CD cavity. In order to test the PVA/CD microsphere inclusion ability, the microspheres were packed in a glass column and the liquid chromatographic behaviour by isocratic elution of different drugs or typical organic compounds, taken as model drugs, was investigated.