Separation of chondroitin sulfate and hyaluronic acid fragments by centrifugal precipitation chromatography

Centrifugal precipitation chromatography (CPC) was applied for the first ti me to the separation of fragments of chondroitin sulfate (ChS) and hyaluron ic acid (HA). The separation was performed using a gradient elution system between ethanol and water since solubility of these biopolymers highly depe nds on the concentration of ethanol in aqueous solution. ChS and HA were ea ch eluted into several peaks through a flow-through UV detector at 275 nm, despite they have almost no absorbance at this wavelength in an aqueous sol ution. The separation was also confirmed by redissolving the dried fraction in water and measuring the absorbance at 210 nm. These results suggest tha t the CPC system can detect small precipitates of these biopolymers by ligh t scattering at 275 nm. The separated fragments of biopolymers are not easi ly characterized because no suitable analytical method is available for ide ntification of these compounds. However, the overall results demonstrate th at CPC may be a useful separation of biopolymers such as glycosaminoglycans which quantitatively produce precipitates in an organic solvent mixture. ( C) 2001 Elsevier Science B.V. All rights reserved.