CHEMICAL MICRODETERMINATION OF HEPARIN IN PLASMA

A new, simple, and highly sensitive method for the determination of he parin has been established. Heparin was first converted into unsaturat ed disaccharides through the action of heparin lyases I, II, and III. A major trisulfated unsaturated disaccharide product results, consiste nt with structural analysis of a number of pharmaceutical heparins usi ng one-and two-dimensional H-1 NMR spectroscopy. This disaccharide was analyzed by HPLC using fluorometric postcolumn derivatization. The co rrelation between the amount of this trisulfated unsaturated disacchar ide and anticoagulant activity of heparin as measured by anti-IIa was determined. The analysis of these pharmaceutical heparins showed a lin ear correlation between both HPLC and bioassay. This HPLC method was t hen applied to a pharmacokinetic study of heparin intravenously admini stered to rabbits. (C) 1997 Academic Press.