Experimental analysis of the biochemical structure of mouse liver heparin sulfate

[Objective] To evaluate a biochemical analysis method for heparin sulfate and to experimentally analyze the disaccharide composition and chain length of mouse liver heparin sulfate. [Methods] Extraction and purification of the heparan sulfate were adapted by taking the advantage of its physical and chemical properties. The purified samples were labeled with 2-Aminobenzomide (2-AB) followed by high performance liquid chromatography (HPLC) analysis. Resultant chromatography was compared with the disaccharides standards to get the disaccharide composition and amount compared with the molecular standards to know the molecular weight. [Results] Similar disaccharide composition and disaccharide amount were obtained by digesting and analyzing the glycansaminoglycans purified from the mouse liver. The 0S disaccharide was (7.01±0.81) pmol/mgdry weight,around 58.27% of the total disaccharides,next was NS disaccharide, (3.47±0.99) pmol/mg, 28.27% of the total disaccharides,following was 6S disaccharide[(0.85±0.99) pmol/mg, 6.87%], Tris disaccharide[(0.50±0.33) pml/mg, 3.85%], S1 disaccharide(0.21 pmol/mg, 1.65%) and S2 disaccharide (0.14 pmol/mg, 1.1%). The total disaccharide amount of heparin sulfate was (12.17±2.44) pmol/mg. The molecular weight of heparin sulfate chain was around 28 kD. [Conclusion] High purity heparin sulfate can be obtained by using the extraction and purification methods introduced in this article. The reproducible heparan sulfate disaccharide composition and chain length were obtained by analyzing the purified samples indicating the methods are suitable for examining the heparin sulfate biochemical changes in liver diseases.