Johannes Basten

HPLC Detection of Sulphated Carbohydrates with a New Detection Method, that is Based on Measurement of Optical Activity by a Polarized Laser (ChiramonitorR)

ABSTRACT The synthesis of sulphated pentasaccharides with heparin-like activity1,2,3 proceeds through many chemical steps of protection and deprotection. As long as the pentasaccharide is fully protected with hydroxyl protecting groups such as benzylic and ester groups, the purity of intermediates can be determined by TLC. During deprotection and sulphation procedures, this determination of the purity with TLC becomes more difficult and reversed phase HPLC turns out to be a better chromatographic technique. The purification and purity control of the deprotected and sulphated pentasaccharide I (ORG31540/CY234) is performed on ion-exchange columns with a NaCl gradient as a mobile phase and UV-detection at 215 nm.