Reserch On The Extraction, Purification And Anticoagulation Of Dermatan Sulfate From Chitterlings

Dermatan sulfate is a kind of sulfating glycosaminoglycans, play an important role in many of the physiology, pathology process. Pig small intestine is an important source of commodity dermatan sulfate, and research in dermatan sulfate from Pig small intestine are very limited whether at home and abroad at present. This paper main aiming at research in extracting dermatan sulfate from pig small intestine by enzymatic hydrolysis, and purify by macroporous resin. The results are as follows:1. optimization extraction process of dermatan sulfate by using enzymaticthis paper uses the protamex to extract dermatan sulfate from pig small intestine,regard hydrolyzed time,tempareture,enzyme additive amount,pH as the influence factors of extraction.And take exatraction as evaluation indexes.Determine the optimal extracting parameters by orthogonal test, then get the final optimized conditions:Heat the50ml mucosa diluent to55℃,Mediation of pH to7.5,Add0.9g of protamex, keeping mix7.5h in low speed Under constant temperature. then use absolute ethyl alcohol of2times the volume to precipitate it.2. optimization purification process of dermatan sulfate by using macroporous resinBy the resin screening test,through five different resin as BS-Ⅱ, D208, D254, D204, D241, determine resin D208to apply to purify dertaman sulfste. Through the static adsorption of macroporous resin D208type and analytical test,Get the best static adsorption analytical conditions as:the configuration of the DS samples, initial concentration of0.15mg/ml,100ml configured sample liquid, mediation of pH to9.0, add10g resin D208accurately say, in the table of55℃in adsorption at200r/min of oscillation speed7h can achieve the optimal adsorption rate. With the volume fraction of95%, after the completion of adsorption of ethanol on the resin. Through macroporous resin D208type dynamic adsorption and resolution test, studied the purification of skin pigment sulfate liquid concentration, all the best of the sample flow rate, eluent, flow rate, get purification of sulfuric acid, the optimal conditions of skin pigment, weigh and resin D208type5g has pretreatment with wet packing column, concentration of1.0mg/ml of sulphuric acid on skin pigment sample liquid column, of1.0ml/min flow velocity for dynamic adsorption; After adsorption, the adsorption column leaching with distilled water first, again with95%ethanol solution, velocity resolution of1.0ml/min. Final lyophilization after purification of skin pigment, sulfate purity is73.96%, protein concentration was0.84%.3. the anticoagulant actinity of the dermatan sulfate from pig small intestinethe anticoagulant actinity of the dermatan sulfate from pig small intestine was invested in terms of effection of dermatan sulfate on coagulant time and prothrombin time with heparinum as control group. The results showed that the dematan sulfate has significant effect on coagulant time and has no effect on prothrombin time which indicating that the dematan sulfate mainly effect on intrinsic pathway.