determinationofmodificationdegreeofsuccinylatedgelatinbysizeexclusionchromatographycoupledwithmultianglelaserlightscatting

The clinical effectiveness of succinylated gelatin as a plasma substitute depends on its molar mass and free amino modification degree. In this study, a new method was developed to determine the modification degree of succinylated gelatin. Gelatin and succinylated gelatin were derived with trinitrobenzenesulfonic acid (TNBS). The molar masses of gelatin and succinylated gelatin were determined before and after TNBS-derivation by size exclusion chromatography coupled with multi angle laser light scatting (SEC-MALLS). The modification degree was obtained by determining the free amino groups of gelatins before and after succinylation. Human serum album in was used as a model protein to validate the method. The determined value was consistent with the calculated value. Actual samples were analyzed with the developed method. The result indicates that the mean modification degree of succinylated gelatins is 38%. This research indicates that size exclusion chromatography multi angle laser light scatting coupled with the trinitrobenzenesulfonic acid derivation is a possible method to determine the modification degree of succinylated gelatin