Determination of unsaturated disaccharides produced from chondroitin sulfate in urine using high performance liquid chromatography with fluorometric post column derivatization / 中国地方病学杂志

ABSTRACT

Objective To explore the effectiveness of the unsaturated disaccharides derived from chondroitin sulfate(CS)in urine using post column derivation fluorometric method of high performance liquid chromatography (HPLC).Methods Urine samples were prepared with the centrifugal flirtation and the routine precipitation methods separately,then digest the sample with CS lyses,use the post column derivation method of HPLC to fluorometric determine the unsaturated disaecharides(△Di-0S,△Di-4S and △Di-6S),compare the recovery differences of the CS standard in the urine pretreated by the two methods.Results The HPLC system ran well,the lowest detection limit of the method was 10 ng.When the urine was pretreated by the centrifugal flirtation method,the recovery of △Di-0S,△Di-4S and △Di-6S produced from the CS standard were 86.93%,87.28%and 85.03%,respectively,while the recovery of the three disaccharides in routine precipitation method were 75.09%,72.96%and 77.70%merely.Conclusions The HPLC post column derivatization fluorometric method is comparatively sensitive,it can be used to determine unsaturated disaccharides decomposed by CS.In the pretreatment procedure of the urine,the centrifugal flirtation method was superior to the routine precipitation method,it can be used as microdetermination for the unsaturated disaccharides.