Method for extracting unfractionated heparin from camel intestinal mucosa

A method of making heparin using mucosa obtained from a camel, the method comprising:

• adding calcium chloride to the mucosa to obtain a first mixture, wherein the calcium chloride calcifies the mucosa;
• removing excessive calcium chloride from the first mixture by adding sodium carbonate to create a second mixture;
• adding bleach to the second mixture to obtain pure mucosa, wherein the bleach comprises about 1% to about 4% w/v H2O2;
• transferring the pure mucosa from the second mixture to a container;
• raising the temperature of the pure mucosa;
• adding sodium chloride to the pure mucosa to create a third mixture, wherein the sodium chloride is 5-15% w/v and iodine free;
• adjusting the pH of the third mixture to a value of about 8 to about 10;
• raising the temperature of the third mixture to a temperature ranging from about 40° C. to about 60° C.;
• adding protease to the pure mucosa to obtain a fourth mixture and leaving the fourth mixture to react for about 2 hours to about 5 hours;
• adding calcium chloride to the fourth mixture to create a fifth mixture, wherein the calcium chloride is a 2-8% solution;
• raising the temperature of the fifth mixture to about 80° C. to about 100° C. and leaving the third mixture to react for about 0.5 hours to about 2 hours;
• adding sodium carbonate to the fifth mixture to obtain a solution and allowing the solution to react for about 0.5 hours to about 2 hours, wherein the sodium carbonate is a 2-5% solution;
• allowing the solution to cool;
• centrifuging the cooled solution and collecting a first clear solution;
• adjusting a pH of the first clear solution, wherein the pH of the first clear solution is adjusted to a value ranging from about 9 to about 12;
• resting the first clear solution for about 0.5 hours to about 2 hours;
• centrifuging the first clear solution and collecting a second clear solution;
• adjusting a temperature of the second clear solution to about −10° C.;
• adjusting the pH of the second clear solution to a value ranging from about 1 to about 4 and resting the second clear solution for about 0.5 hours to about 2 hours;
• centrifuging the second clear solution and collecting a third clear solution;
• adjusting a pH of the third clear solution to a value ranging from about 9 to about 12;
• adding 1-4% w/v H2O2 to the third clear solution to form a sixth mixture and maintaining the temperature of the sixth mixture at about 20° C. to about 30° C. for a period of time ranging from about 30 hours to about 50 hours;
• centrifuging the sixth mixture and collecting a fourth clear solution;
• adjusting a pH of the fourth clear solution to a value ranging from about 9 to about 12, wherein the pH of the fourth clear solution is adjusted by adding 1-3% w/v H2O2;
• maintaining a temperature of the fourth clear solution at a temperature of about 30° C. to about 50° C. for about 10 hours to about 20 hours;
• collecting a fifth clear solution through centrifugation and filtration;
• adding a 10-20% w/v iodine free sodium chloride to the fifth clear solution to obtain a seventh mixture;
• keeping the seventh mixture overnight;
• diluting the seventh mixture with water 5 to 10 times to obtain a sixth mixture;
• adding 0.5 volume of pure ethanol into the seventh mixture;
• adjusting a temperature of the seventh mixture to about −30° C.;
• keeping the seventh mixture overnight;
• and
• collecting and drying a pure precipitate to obtain a pure unfractionated heparin (UFH).