This project was designed to use poultry waste to isolate and characterize collagen type II

from its trachea. Collagen type II is being used along with condroitin sulfate and

glucosamine for the treatment of osteoarthritis and is also available as a neutraceutical

product in the market. For project purpose, trachea of slaughtered broiler birds were

collected from the market and after removing adhering tissue and debris, it was then

washed thoroughly first with distilled water and then with deionized water. Tracheal

cartilage was then cut into small pieces and defattened with chloroform: methanol (2: 1

v/v) solution. After this, the cut pieces were properly cleaned with deionized water. 0.5%

Pepsin solution in 0.5 M acetic acid was prepared. Cartilage was then hydrolyzed by the

already prepared 0.5 % pepsin (in 0.5 M acetic acid) at 4 ° C for 48 hours. The extract
was then separated from the tracheal pieces and the viscous solution obtained was

centrifuged at 12000 rpm for 1 hr at 4 "c. Now the collagen was expected to be in the

supernatant which was salted out by adding NaCI to a final concentration of 2.5M and

kept for almost 12-16 hrs. This collagen was again centrifuged at 12000 rpm for 1 hr at

4 C. The obtained collagen pallet was redissolved in 0.5 M acetic acid and then it was

dialyzed against 0.1 M acetic acid followed by dialysis with distilled water. The sample

after dialysis was put in petri dishes and kept in freezer for overnight to let it be prepared for

lyophilization. The frozen collagen sample was then lyophilized. After lyophilization, the sample

gave an appearance of a white mesh. This sample was reconstituted in PBS with pH 8 to run it on

SDS-PAGE. The procedure of SDS-PAGE in non reducing conditions was adopted for the

characterization of collagen type II in the sample. The description of results of SDS-PAGE is given

below:

Lane M contains protein markers of different molecular weight. Lane 1, 2 and 3 contains

samples at different steps of the whole procedure showing clear bands of collagen type II.

Lane 4 contains lyophilized sample of collagen type II showing the thickest band (alpha

chain of collagen type II). In this research, poultry waste has been used for making health improving

product. As in our country poultry is used in bulk quantity so if its waste might be used in any

medicinal product then it might not only be useful but also economical for such a developing country

as ours. Another thing is that as this collagen Type II has been extracted from poultry trachea, it

shows that tracheal cartilage is a rich source of such collagen type. Collagen Type II is used in the

cure of arthritis especially rheumatoid arthritis so through this research, it has been made clear that

poultry waste can be utilized in a positive way in medicinal industry and also that collagen Type II

acts as an effective neutraceutical.