MANUFACTURED PRODUCT USING AND COLLAGEN SOLUTION MANUFACTURING METHOD AND COLLAGEN SEPARATION METHOD OF ANIMAL TISSUE

A method for separating collagen from various animal tissues is provided to be able to efficiently obtain the collagen from osseous tissue, cartilage tissue, skin tissue and tendon with improved product quality and reliability. The method for separating collagen from osseous tissue of pig comprises the steps of: (a) washing the osseous tissue of pig with distilled water, alcohol, and acetone; (b) after cutting the osseous tissue into doughnut-shaped tissues, storing it at a temperature of -20 deg.C; (c) pulverizing the osseous tissue into powders having a size of 1-500 micrometers; (d) washing the powdered osseous tissue with alcohol and distilled water; (e) treating the washed osseous tissue powder with pepsin, where the ratio of the tissue to pepsin is 10-50:1 and the pepsin is dissolved in 0.1N HCl, 2-5 times for 3-7 days; (f) after centrifuging the pepsin-treated osseous tissue powder(12,000g, 30 minutes, 4 deg.C), storing separated supernatant and allowing precipitating material to repeat the step(e); (g) treating the supernatant with 0.5-0.8M NaCl for 4 hours to 1 day at a temperature of 4 deg.C; (h) after centrifuging it(12,000g, 30 minutes, 4 deg.C), removing supernatant therefrom and collecting precipitating materials therefrom at a temperature of 4 deg.C; (i) after dissolving the precipitating materials in a solvent to be neutral, adding NaCl thereto to obtain the solution at final NaCl concentration of 1.6M; (j) after leaving the solution obtained from the step(i) at a temperature of 4 deg.C for 4 hours to 1 day, centrifuging it to remove precipitate therefrom and collect supernatant therefrom; (k) after adding NaCl to the collected supernatant to obtain a solution at a final NaCl concentration of 2.6M, leaving it at a temperature of 4 deg.C for 4 hours to 1 day; (l) after centrifuging the solution to remove supernatant therefrom, washing precipitate with 95% alcohol 1-2 times and re-suspending it in distilled water; (m) after completely re-suspending the solution obtained from the step(l) with adding 1ml of 1N HCl to 100ml of the solution, titrating it to be neutral at a temperature of 4 deg.C; (n) leaving the titrated solution at a temperature of 30-37 deg.C for 4 hours to 1 day and then centrifuging it; and (o) re-suspending the precipitated collagen in ddH2O or phosphate buffer saline(PBS) at a concentration of 1-30mg/ml and then storing it at a temperature of 4 deg.C.