Study On The Extration Technology Of Chondroitin Sulfate

Chondroitin sulfate, relative molecular mass from 10,000 D to 50,000 D, is an acidic polysaccharide composed of disaccharide units of glucuronic acid and hexosamine. Chondroitin sulfate distributing in various animal tissues has been proven to posses various biological activities such as adjusting immunity and fat metabolism,preventing infection,reducing the risk of heart diseases. Therefore, chondroitin sulfate has great application potentials in fields of food,medicine and cosmetic.The research used porcine laryngomalacia as testing material, alkali extraction method, alkali-single enzyme extraction method, alkali-double enzyme extraction method and ultrasonic assistant extraction method were used in extraction of chondroitin sulfate. The purity (hexosamine) and the production yield have been investigated through tests.1.In the process of alkali extraction, some factors influence the effect of the production yield including ratio of cartilage quality and lye liquor, lye consistency, reaction temperature, reaction time have been investigated through orthogonal tests. Based on the studies, the optimization technology conditions were ratio of cartilage quality and lye liquor 1:6, reaction temperature 40℃, lye consistency 6%, reaction time 6h, production yield achieved more than 17.01% and purity 70.32%.2.In the process of alkali-single enzyme extraction, the study used papain proteinase to hydrolyze. Some factors influence on the production purity including papain additive amount, hydrolysis temperature, pH, hydrolysis time have been investigated through single factor optimizing tests and orthogonal tests. The optimization technology conditions were pH 6.8, hydrolysis temperature 40℃, the papain additive amount 1.2:1000, hydrolysis time 60min, the purity achieved 79.32% production yield 18.56%.3.In the process of alkali-double enzyme extraction, based on the alkali-single enzyme extraction studies, used pepsin to made further hydrolyze. hydrolysis temperature, pH, pepsin additive amount, hydrolysis time have been investigated through single factor optimizing tests and orthogonal tests. The optimization technology conditions were pH 5.9, hydrolysis temperature 40℃, pepsin additive amount 1.4:1000, hydrolysis time 100 min, the purity achieved 82.76%, production yield 20.93%.4.In the process of ultrasonic assistant extraction, based on the alkali-double enzyme extraction studies. The optimization technology condition were ultrasonic treatment time 139 min, power 491 W, pH6.8, papain hydrolysis temperature 40℃, papain additive amount 1.2:1000, hydrolysis time 60min, pH 5.9, pepsin hydrolysis temperature 40℃, pepsin additive amount 1.4:1000, hydrolysis time 100min, the purity achieved 86.06% and production yield 22.56%.5.Compared with physical and chemical indexes, sensory indexes and microbiology indicators of four methods. The results showed that all indexes of ultrasonic auxiliary alkali-double enzymatic products were superior to other methods, the purity was the highest achieved 86.06% and more than export standard.6.The products of ultrasonic auxiliary extraction purified with ethanol.When the concentration of chondroitin sulfate solution 18% (w/v), NaCl 3% (w/v), pH 6.8, concentration of ethanol system 70% (v/v), repeated 4 times, the products were slightly salty white power. Protein 2.78%, hexosamine 31.98%, chondroitin sulfate 90.17%. Compared with the before purified 86.06% (4.11%increased). The moisture 6.30%, clarification 0.035, pH 7.01. Others indexes also meet with standard.7.In order to further verificated the purity of alkali-double enzymatic extraction, ultrasonic auxiliary extraction and purified, Used the infrared spectrum test. Results showed that alkali–double enzymes method, ultrasonic auxiliary extraction and standard, The corresponding absorption peaks existed, but there was little impurity peak. Compared with the standard, purification products similar to standard achieved high purity.Ultrasonic auxiliary extraction was a short-time consuming, easy operation and high purify method.