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J. Env. Bio-Sci., 2018: Vol. 32 (1): 45-47 ISSN 0973-6913 (Print), ISSN 0976-3384 (On Line)

A PRELIMINARY STUDY ON DEVELOPING COLLAGEN SHEET AS BIOMATERIAL

USING ACID SOLUBLE COLLAGEN EXTRACTED FROM CHICKEN SKIN
K. Arunmozhivarman*, Robinson J.J. Abraham, V. Appa Rao, M. Parthiban1, R. Narendra Babu,
S. Ezhilvelan and C. Vasanthi
Department of Livestock Products Technology (Meat Science), Madras Veterinary College
Tamil Nadu Veterinary and Animal Sciences University
Chennai - 600 007, Tamil Nadu, India.
1
Department of Animal Biotechnology,Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University
Chennai - 600 007, Tamil Nadu, India.
[Corresponding author*: arunmozhivet@gmail.com]

Received: 09-04-2018 Accepted: 20-04-2018

The objective of this research work is to extract collagen from the chicken skin and also make an attempt to develop a collagen
based sheet as a biomaterial. Consumption of broiler meat level is always high when compared to beef, mutton and chevon
because of low cost and no religious taboo in the society. This has lead to increase in slaughter rate and generation of large
amount of by-products. In poultry meat retail shop the by-products wastes produced are mostly disposed of into water bodies, free
lands which ultimately leads to environmental pollution. Hence in this study, the chicken skin was utilized to extract collagen using
acetic acid solubilisation method. The extracted collagen was lyophilized, evaluated for its characteristic quality parameters and
collagensheet was prepared with addition of chitosanas biomaterial.
set,
Key words: Collegen sheet, biomaterial, Chicken skin

Collagen a fibrous structural protein can be resorbed into the
body, a non-toxic and biocompatible, generates only minimal
immune response (low antigenicity), and was excellent source
for attachment, biological interaction with cells (biocompatibility)
and proteolytic degradation pathway (Pati et al., 2012).
Collagen has been extracted from by-products of food animals
such as cattle, pig as well as marine sources which have
been widely used in food, pharmaceuticals and cosmetic
industries.In India, large amount of chicken skin are generated
as by-product during poultry meat processing. Avian collagen
is considered as alternative source for various skin care and
medical application instead of conventional bovine and porcine
and aquatic collagen (Lin et al., 2013). The present day
consumers are more health conscious most of them started
avoiding consuming fatty foods and so in our country there is
increase in market for skin less chicken meat which results in
generated more chicken skin as a by-product. Hence, this
study was conducted with the objectives of extraction and
characterization of collagen from chicken skin and to develop
collagen sheet for biomaterial application.
MATERIAL AND METHODS
Chicken skin were procured from the local market and brought
to the laboratory immediately. Each trial was carried out with
NAAS Rating (2017)-4.43
50 gram of minced skin and collagen extraction process was
done at 4°C in walk-in chiller. Collagen from the chicken skin
was extracted using acetic acid solubilisation method (Liu et
al., 2001) with modification. Pretreatment chicken skin was
done 0.1N NaOH, 20% ethanol to remove non-collagenous
protein and fat each step for 24 hours. Then samples were
treated with 0.5M acetic acid in the ratio of 1:30 (w/v).Both the
extract and super solids were homogenized and then filtered
using double cheese cloth to obtain the filtrate. Collagen from
the filtrate was precipitated by using 2.6 M NaCl. The resulted
precipitates were collected by centrifugation at 10000 g for 6
minutes at 4°C. The precipitates were dissolved in 0.5 M acetic
acid and dialyzed against 0.1M acetic acid and distilled water
followed by lyophilization to obtain dried collagen.Colour of
collagen was measured using Hunter colour lab mini scan XE
plus spectro colorimeter and the colour was expressed as
Lightness (L*), Redness (a*), and yellowness (b*). The Protein
com position of collagen (AOAC,1995); Am ino acid
composition; FTIR analysis were also done as per standard
protocol.The collagen sheet was according to the method of
Bama et al. (2010) with proper modification. The 2% collagen
and chitosan solution were prepared by dissolving in 0.5M
acetic acid and mixed thoroughly in a magnetic stirrer to obtain
a uniform protein and polysaccharides solution. Ten to fifteen
 ARUNMOZHIVARMAN, ABRAHAM, APPARAO, PARTHIBAN, NARENDRA BABU, EZHILVELAN AND VASANTHI (46)

ml of the solution taken and transferred into petridisc and frozen Analysis of amino acid (Table 1) revealed that extracted chicken
at -20°C and then lyophilized to obtain collagen sheet (Fig. 2). skin collagen had high content of glycine (7587nano moles/
RESULTS AND DISCUSSION ml) followed by histidine, glutamic acid and arginine. Less
amount of lysine and isoleucine in this study indicates that
The proximate analysis of acid soluble collagen revealed that effective removal of telopeptide regions from collagen (Knott
it has high percentage of protein (90.23±0.56) which indicates and Bailley, 1998).
the efficiency of extraction method.Instrumental colour analysis
showed the extracted collagen had higher value for lightness Study of changes in the secondary structure of collagen using
(L*= 89.18±0.87) and less value in yellowness (b*= 9.57±0.41), FTIR revealed that the amide bands A, B, I, II and III for extracted
redness (a*= -0.81±0.15). The analysis of colour revealed that collagen were occurred at 3305.73 cm-1, 2922.85 cm-1, 1633.47
the extracted collagen is whitish grey in colour indicating that cm -1, 1547.91 cm -1, 1237.03 cm -1respectively (Fig.1).The
efficient removal of fat and haemoglobin pigment (Huda et al., absorption of amide III was found at 1237.03 cm -1 and which
2013) during the process of extraction. indicates the existence of collagen helical structure (Liu et
Table 1: Amino acid profile collagen
al., 2007).
Amino acids Nanomoles/ml
Aspartic acid 392 Chvapil et al. (1973) reported that collagen could be processed
Glutamic acid 2426 into a number of forms such as sheets, sponges, powders,
Serine 1682
injectable solutions and dispersions, all of which have found
Histidine 5274
use in medical practice.Based on the results on molecular
Glycine 7587
Threonine 1935
and structural characteristics of extracted collagen revealed
Arginine 2190 that it can be used in biomaterial application.The prepared
Alanine 1187 collagen sheet in this study can be used for clinical purposes
Tyrosine 373 with further studies.
Methionine 767
Valine 1190
Phenylalanine 287
Isoleucine 91
Leucine 1068
Lysine 196
3305.73

3079.33

2922.85
2853. 50

2347.72

2090.91

1742.83
1633.47
1547.91
1453.78
1402.88
1336.63
1237.03
1158.01
1080.69
1026.64

875.75

718.60
651. 58
1.4
1.2 1.0
AbsorbanceUnits
0.6 0.8
0.4
0.2
0.0

3500 3000 2500 2000 1500 1000 500

Wavenumber cm-1
Fig.1: FTIR spectrum analysis of acid soluble collagen
(47) A PRELIMINARY STUDY ON DEVELOPING COLLAGEN SHEET
Fig. 2: Collagen sheet.
ACKNOWLEDGEMENTS
The authors are thankful to the Director, AICRP on Post Harvest
Engineering Technology and Tamil Nadu Veterinary and Animal
Sciences University for providing necessary funds and facilities
to carry out research work.
REFERENCES
1. AOAC (1995). Official methods of analysis (16thed). Washington,
DC: Association of Official Analytical Chemists.
2. Bama, P., Vijayalakshami, M., Jayasimman,R., Kalaichelvan,P.,
Deccaraman, M. and Sankaranarayanan, S.(2010). Extraction of
collagen from cat fish (Tachysurus maculatus) by pepsin digestion
and preparation and characterization of collagen chitosan sheet.Int
Jour. Pharm. Sci. 2(4): 133-137.
3. Chvapil. M., Kronenthal, R.L. and Winkle, W.V. (1973). Medical and
Surgical Applications of Collagen. Int. Rev. Connect. Tissue. Res.,6:
1-61.
4. Huda, N., Seow, E. K., Normawati, M. N. and NikAisyah, N. M.
(2013). Preliminary Study on Physicochemical Properties of Duck
Feet Collagen. Inter. Jour. Poult. Sci.,12: 615-621.
5. Knott, L. and Bailley, A.J. (1998). Collagen cross-links in mineralizing
tissues: A review of their chemistry, function, and clinical relevance.
Bone.,22: 181-187.
6. Lin, C.W., Loughran,M., Tsai, T.Y. and Tsai, S.W. (2013). Evaluation
of convenient extraction of chicken skin collagen using organic
acid and pepsin combination. Jour. Chin. Soc. Anim. Sci., 42(1):
27-38.
7. Liu, D.C., Lin, Y. K. and Chen, M.T. (2001). Optimum condition of
extraction of collagen from chicken feet and its characteristics.
Asian Austral. Jour. Anim. Sci.,14: 1638-1644.
8. Liu, H., Li, D. and Guo, S. (2007). Studies on collagen from the skin
of channel catfish (Ictalurus punctaus). Food Chem.,101: 621-
625.
9. Pati, F., Datta, P., Adhikari, B.,Dhara, S., Ghosh, K. and Mohapatra,P.
K. D.(2012). Collagen scaffolds derived from fresh water fish origin
and their biocompatibility. Jour. Biomed. Mater. Res. part A. 100(A):
1068-1079.