Method for purifying high molecular weight hyaluronic acid

A method for purifying high molecular weight hyaluronic acid, comprising the steps of,

• disinfecting one or more raw hyaluronic acid sources;
• placing the disinfected source in water with an antiseptic to form a solution;
• mixing said solution until the solution forms an extract having a viscosity of at least about 500 cps;
• straining said extract from said solution and treating with NaCl;
• centrifuging said treated extract and adding ethanol to form a precipitate;
• dissolving said precipitate in water;
• extracting enzyme inhibitors from said dissolved precipitate with chloroform and centrifuging to produce an aqueous portion;
• adding one or more protein hydrolyzing enzymes to said aqueous portion in a reactor;
• adding to the reactor a solution of about 2% cetyl pyridinium chloride (CPC) and about 0.3M NaCl to precipitate non-hyaluronic acid glycosoamino glycans;
• filtering reactor contents through at least one membrane filter to remove precipitants;
• filtering membrane filtered solution through at least one diafilter having a molecular weight cutoff of about 30 kDa to remove low molecular weight contaminants;
• and
• precipitating diafiltered solution and isolate purified hyaluronic acid precipitate.