Preparation and purification of corticotrophin

Claim 1

A METHOD FOR OBTAINING HIGH YIELDS OF HIGHLY ACTIVE CORTICOTROPHIN FROM ANIMAL PITUITARY GLANDS, COMPRISING; EXTRACTING SAID GLANDS WITH A SOLVENT MIXTURE CONTAINING ABOUT 65 TO 80 PERCENT OF A ORGANIC SOLVENT SELECTED FROM THE GROUP CONSISTING OF ACETONE AND ETHANOL AT A PH OF ABOUT 1.5 AND AT THE REFLUX TEMPERATURE OF THE SOLVENT; SEPARATING THE RESULTANT SOLUTION FROM THE PROTEINACEOUS RESIDUE; DECOLORIZING THE RESULTANT SOLUTION AT A PH BELOW ABOUT 2.0 WITH DECOLORIZING CHARCOAL TO THEREBY SELECTIVELY REMOVE BLOOD PIGMENTS; ADDING AN AQUEOUS SOLUTION OF A WATER-SOLUBLE ZINC SALT TO THE DECOLORIZED EXTRACT; GRADUALLY RAISING THE PH OF THE RESULTANT SOLUTION TO ABOUT 4.45 TO PRECIPITATE GROWTH HORMONE AND OTHER PRINCIPLES OF PITUITARY; SEPARATING THE SUPERNATANT SOLUTION FROM THE PRECIPITATE; GRADUALLY INCREASING THE PH OF SAID SOLUTION TO ABOUT PH 5.9 TO PRECIPITATE CRUDE CORTICOTROPHIN AS A ZINCE COMPLEX; REMOVING THE ZINC COMPLES OF CORTICOTROPHIN FROM SOLUTION; DISSOLVING SAID CORTICOTROPHIN COMPLEX IN ACID SOLUTION AND ADJUSTING THE PH OF SAID SOLUTION TO ABOUT 3.5 TO 4.5; ADSORBING THE ACTIVE CORTICOTROPHINFROM SAID ACID SOLUTION UPON A MIXTURE OF OXYCELLULOSE AND CARBOXYLIC CATION EXCHANGE RESIN; ELUTING THE ADSORBED CORTICOTROPHIN FROM SAID MIXTURE OF ADSORBENTS WITH A DILUTE ACID SOLUTION; AND PRECIPITATING THE CORTICOTROPHIN FROM SUCH SOLUTION BY POURING SAID ELUATE INTO A COLD ORGANIC SOLVENT WHICH WILL CAUSE PRECIPITATION OF CORTICOTROPHIN.

Claim 2

A METHOD FOR OBTAINING HIGH YIELDS OF HIGHLY ACTIVE CORTICOTROPHIN FROM ANIMAL PITUITARY GLANDS, COMPRISING; EXTRACTING SAID GLANDS WITH A SOLVENT MIXTURE CONTAINING ABOUT 65 TO 80 PERCENT OF A ORGANIC SOLVENT SELECTED FROM THE GROUP CONSISTING OF ACETONE AND ETHANOL AT A PH OF ABOUT 1.5 AND AT THE REFLUX TEMPERATURE OF THE SOLVENT; SEPARATING THE RESULTANT SOLUTION FROM THE PROTEINACEOUS RESIDUE; DECOLORIZING THE RESULTANT SOLUTION AT A PH BELOW ABOUT 2.0 WITH DECOLORIZING CHARCOAL TO THEREBY SELECTIVELY REMOVE BLOOD PIGMENTS; ADDING AN AQUEOUS SOLUTION OF A WATER-SOLUBLE ZINC SALT TO THE DECOLORIZED EXTRACT; GRADUALLY RAISING THE PH OF THE RESULTANT SOLUTION TO ABOUT 4.45 TO PRECIPITATE GROWTH HORMONE AND OTHER PRINCIPLES OF PITUITARY; SEPARATING THE SUPERNATANT SOLUTION FROM THE PRECIPITATE; GRADUALLY INCREASING THE PH OF SAID SOLUTION TO ABOUT PH 5.9 TO PRECIPITATE CRUDE CORTICOTROPHIN AS A ZINCE COMPLEX; REMOVING THE ZINC COMPLES OF CORTICOTROPHIN FROM SOLUTION; DISSOLVING SAID CORTICOTROPHIN COMPLEX IN ACID SOLUTION AND ADJUSTING THE PH OF SAID SOLUTION TO ABOUT 3.5 TO 4.5; ADSORBING THE ACTIVE CORTICOTROPHINFROM SAID ACID SOLUTION UPON A MIXTURE OF OXYCELLULOSE AND CARBOXYLIC CATION EXCHANGE RESIN; ELUTING THE ADSORBED CORTICOTROPHIN FROM SAID MIXTURE OF ADSORBENTS WITH A DILUTE ACID SOLUTION; AND PRECIPITATING THE CORTICOTROPHIN FROM SUCH SOLUTION BY POURING SAID ELUATE INTO A COLD ORGANIC SOLVENT WHICH WILL CAUSE PRECIPITATION OF CORTICOTROPHIN.