Abstract: Collagen fiber cross-linked by glutaraldehyde (GCF) was used to study the chromatographic separation of flavanone glycosides. Quercitrin,rutin and robinin were selected as the representativeflavanone glycosides,which contained different groups of glucosyls. The results showed that the hydrogen-bond interactions were responsible for the adsorption of flavanone glycosides by collagen fiber in high concentration aqueous ethanol,while the hydrophobic interactions dominated the adsorption when the concentration of ethanol was low. The mixture of quercitrin,rutin and robinin can be separated on GCF column by stepwise elution of 90%70%-50% aqueous ethanol. Their purities were 99.39%,95.32% and 94.23%,while the recoveries can be reached to 89.25%，80.36% and 80.49%,respectively. The GCF column was repeatedly used for 5 times without significant reduction of separation capability,indicating the excellent reusability of the GCF column.

Key words: collagen fiber, separating material, flavanone glycosides, chromatographic separation