Extraction And Structural Characterization Of Glycosaminoglycans From Different Swine Organs

Glycosaminoglycans are the glycan part of proteoglycans which can regulate avariety of growth factor signaling pathway. Glycosaminoglycans play multiple rolesin development, health and disease by interacting with proteins. The most commonsource of medicinal glycosaminoglycans is pig tissue, but the study of comparison ofdifferent glycosaminoglycans that extracted from several pig organs are still veryfew.Therefore, this thesis mainly aims at the comparative study of extraction andseparation of glycosaminoglycans from six different pig tissues,as well as thecomposition, content and structure of glycosaminoglycans.In this paper,using pig tissues (pig heart, liver, small intestine, pig lung, kidney,pig small intestinal mucous membrane)as raw material, with acetone andmethanol/chloroform respectively to degrease, extract the GAGs with industrialprotease under the conditions of4h and16h respectively, then isolate polysaccharidewith industrial resin finally use alcohol sinking method to get samples of raw GAGs,the following is the yield of each sample: H4:0.41%; H16:0.43%; Li4:1.2%;Li16:1.05%; Si4:0.52%; Si16:0.55%; Lu4:0.46%; Lu16:0.57%; K4:0.79%;K16:0.39%.The data shows that under the same extraction conditions, differentdigestion time or organizational sources of glycosaminoglycans both can lead todifferent yield rate.With cellulose acetate membrane electrophoresis, sulfuric acid-carbazolemethod, alcian blue method and high performance liquid chromatography (HPLC)method, we analyze the crude polysaccharide samples. The highest content ofglycosaminoglycans is H16samples, the lowest is Li4samples,as to uronic acidcontent the highest is Lu4h, the lowest content is Li4h. Because of heparin andheparan sulfate containing N-acetyl glucosamine，chondroitin sulfate and dermatansulfate containing N-acetyl galactosamine, thus glcN/galN can represent HP; HS/CS; DS.The HPLC analyze the monosaccharide components of glycosaminoglycan，inspite of the small intestine, HP;HS/CS; DS have a lower ratio in the rest sample of16h which shows that longer reaction time,chondroitin sulfate and dermatan sulfate canbe extracted more completly.In this paper, using high performance liquid chromatography-mass spectrometry(LC-MS) method to detect the disaccharide of the samples that extracted from sixdifferent pig tissues, respectively, tested the idose hyaluronic acid and glucuronic acidof glycosaminoglycans successfully which traditional detection methods can’t do.