extraction, enzymolysis, precipitation and drying, weigh pig throat and nasal cartilage, soak in 5 times of 20g/L (2%) sodium hydroxide solution, stir intermittently, filter when the leachate reaches 5.5 ° Be, extract the filter residue again, and merge the filtrate. Hydrolyze with crude pancreatin at 50 ℃ to remove impurities such as protein, filter to clear, adjust pH6, add ethanol to make the alcohol content reach 75%, and let stand overnight. Centrifugal separation, the precipitation is washed with absolute ethanol, and dried at 60°C to obtain crude chondroitin sulfate.
Porcine larynx and nasal cartilage [20g/L NaOH]& rarr; Extract [pancreatin] & rarr;[50 ℃] enzymolysis solution [75% ethanol] & rarr;[pH6] crude products
Grading separation, 1kg of crude product is added into a mixed solution of 20L of distilled water, 1L of acetic acid and 500g of calcium chloride (anhydrous), stirred until fully dissolved, and the volume number of ethanol is added up to 50%, with precipitation and precipitation, and left to stand overnight. Centrifuge to collect calcium salt precipitate. Then dissolve in a mixed solution of distilled water 40L, acetic acid 1.5L and calcium chloride 100g (anhydrous), stir and dissolve, add ethanol with a volume fraction of 29%, precipitate and centrifuge (precipitate and recover CSA-Ca). In the supernatant ethanol solution, continue to add appropriate amount of ethanol, stir to make it white and permanent turbidity, and let it stand overnight. The next day, add appropriate amount of ethanol to make the concentration reach 39%, and place precipitation for 7 days. Centrifuge, wash with absolute ethanol for precipitation, and dry at 60 ℃ to obtain CSA-Ca. Recovery CSA-Ca of supernatant ethanol solution.
Crude products [water, HAc, CaCl2, ethanol] & rarr;CSA-Ca
Transformation, precipitation, drying, CSA-Ca pure product, add appropriate amount of distilled water to dissolve, adjust pH7, adsorb by strong acidic styrene cation exchange resin 001 × 7, collect effluent, add sodium chloride with a solution volume of 5g/L (0.5%), precipitate with ethanol, and let stand overnight. Centrifuge, wash the precipitation twice with anhydrous ethanol, and dry at 60 ℃ to obtain pure chondroitin sulfate sodium A. The total yield of porcine larynx and nasal cartilage was calculated as 5%.
CSA-Ca[001 × 7 resin (Na type) & rarr;[pH7] effluent (CSA-Na)[NaCl, ethanol] & rarr; Precipitate [60 ℃]& rarr; Pure CSA-Na.