Abstract

Collagen is highly valued both as a food additive and a functional food ingredient. It is generally
extracted by treatments with acid or alkali, enzyme, and microorganisms. However these
methods are generally batch type, time-, energy-, reactant-, and cost-consuming. Extrusion is
widely used in the food industry, and offers many advantages, such as ease of operation,
continuous production, high yield, and little waste. In this study, we developed a novel extrusion-
hydro-extraction (EHE) process for extraction of collagen from tilapia fish scale. Extruded scale
samples had a 2-3times higher protein extraction yield than that of non-extruded scale samples.
All extracts contained hydroxyproline (61-73 residues/1000 residues) and hydroxylysine (5-6
residues/1000 residues) and were identified as type-I collagens by FTIR, SDS-PAGE, and
molecular weight distribution analyses. The physicochemical studies revealed that extracted
collagens could have promising applications in the food, medical, and cosmetic industries.
Copyright © 2015 Elsevier Ltd. All rights reserved.

Isolation and characterization of fish scale collagen from tilapia (Oreochromis sp.) by a novel
extrusion-hydro-extraction process (PDF Download Available). Available from:
https://www.researchgate.net/publication/279520339_Isolation_and_characterization_of_fish_sc
ale_collagen_from_tilapia_Oreochromis_sp_by_a_novel_extrusion-hydro-extraction_process
[accessed Jul 16, 2017].

Isolation and characterization of fish scale collagen from tilapia

(Oreochromis sp.) by a novel extrusion–hydro-extraction process
Chun-Yung Huang

Isolation and characterization of fish scale collagen from tilapia (Oreochromis sp.) by a novel
extrusion-hydro-extraction process (PDF Download Available). Available from:
https://www.researchgate.net/publication/279520339_Isolation_and_characterization_of_fish_sc
ale_collagen_from_tilapia_Oreochromis_sp_by_a_novel_extrusion-hydro-extraction_process
[accessed Jul 16, 2017].
Collagen is highly valued both as a food additive and a functional food ingredient. It is generally
extracted
by treatments with acid or alkali, enzyme, and microorganisms. However these methods are
generally
batch type, time-, energy-, reactant-, and cost-consuming. Extrusion is widely used in the food
industry,
and offers many advantages, such as ease of operation, continuous production, high yield, and
little
waste. In this study, we developed a novel extrusion–hydro-extraction (EHE) process for
extraction of
collagen from tilapia fish scale. Extruded scale samples had a 2–3 times higher protein extraction
yield
than that of non-extruded scale samples. All extracts contained hydroxyproline (61–73
residues/1000
residues) and hydroxylysine (5–6 residues/1000 residues) and were identified as type-I collagens
by
FTIR, SDS–PAGE, and molecular weight distribution analyses. The physicochemical studies
revealed that
extracted collagens could have promising applications in the food, medical, and cosmetic
industries

Analytical Methods
Isolation and characterization of fish scale collagen from tilapia
(Oreochromis sp.) by a novel extrusion–hydro-extraction process
Chun-Yung Huang
Department of Seafood Science, National Kaohsiung Marine University, No. 142, Hai-Chuan
Rd., Nan-Tzu, Kaohsiung 811, Taiwan, ROC
b
Department of Health and Nutrition, Chia Nan University of Pharmacy and Science, No. 60, Sec.
1, Erh-Jen Rd., Jen-Te District, Tainan 717, Taiwan, ROC
article info
Article history:
Received 11 January 2015
Received in revised form 14 June 2015
Accepted 20 June 2015
Available online 22 June 2015
Keywords:
Collagen
Extrusion
Extrudate
Hot water extraction
Tilapia fish scale

abstract
Collagen is highly valued both as a food additive and a functional food ingredient. It is generally
extracted
by treatments with acid or alkali, enzyme, and microorganisms. However these methods are
generally
batch type, time-, energy-, reactant-, and cost-consuming. Extrusion is widely used in the food
industry,
and offers many advantages, such as ease of operation, continuous production, high yield, and
little
waste. In this study, we developed a novel extrusion–hydro-extraction (EHE) process for
extraction of
collagen from tilapia fish scale. Extruded scale samples had a 2–3 times higher protein extraction
yield
than that of non-extruded scale samples. All extracts contained hydroxyproline (61–73
residues/1000
residues) and hydroxylysine (5–6 residues/1000 residues) and were identified as type-I collagens
by
FTIR, SDS–PAGE, and molecular weight distribution analyses. The physicochemical studies
revealed that
extracted collagens could have promising applications in the food, medical, and cosmetic
industries.
Ó2015 Elsevier Ltd. All rights reserved.
1. Introduction
Collagen is the most abundant protein in vertebrates and con-
stitutes about 25% of vertebrate total proteins (Ogawa et al.,
2004). Collagen is able to form insoluble fibers that have high ten-
sile strength and a right-handed triple superhelical rod, consisting
of three almost identical polypeptide chains. Nondenatured colla-
gen can be utilized in cosmetics, biomedical, and pharmaceutical
industry applications. Denatured collagen, known as gelatin, is
water-soluble and finds broad applications in the food, pharma-
ceutical, photographic, cosmetic, and packaging industries
(Ogawa et al., 2004)

Collagen from aquatic by-products is generally obtained by the

following methods: (1) Acid and alkali extraction: The prepared

skin was mixed with NaOH and stirred for 2 h to remove
non-collagenous proteins. The mixture was then washed with
tap water until neutral or faintly basic pH was obtained. The depro-
teinized skin was subsequently demineralized using HCl for 1 h.
Then, the residue was washed thoroughly with tap water until
wash water became neutral or faintly basic in pH. Thereafter, the
demineralized skin was swollen by mixing the skins with acetic
acid and stirred for 15 min. Finally, the swollen skin was washed
thoroughly with tap water until neutral or faintly acidic pH of
wash water was obtained. The swollen skin was mixed with dis-
tilled water at different temperatures (45–75 °C) for various times
(6–12 h). The mixtures were then filtered and collagen was

Tilapia is the most widely cultured fish in Taiwan and mainland

China. Taiwan is an important supplier of whole-frozen tilapia to

the U.S. and also exports high quality chilled tilapia fillets to the
Japanese sashimi market. Numerous fish scales which are regarded
as waste are produced from fish fillet processing factories.
Generally, collagen extracted from fish scale has a less malodorous
smell than that from fish skin and bone. Fish scale is known com-
posed of collagen (generally type I) and hydroxyapatite, and both
are tightly linked together (Ikoma, Kobayashi, Tanaka, Walsh, &
Mann, 2003) and difficult to separate (Stepnowski, Ólafsson,
Helgason, & Jastorff, 2004).

Biological Safety of Fish (Tilapia) Collagen

Abstract
Marine collagen derived from fish scales, skin, and bone has been widely investigated for
application as a scaffold and carrier due to its bioactive properties, including excellent
biocompatibility, low antigenicity, and high biodegradability and cell growth potential. Fish type
I collagen is an effective material as a biodegradable scaffold or spacer replicating the natural
extracellular matrix, which serves to spatially organize cells, providing them with environmental
signals and directing site-specific cellular regulation. This study was conducted to confirm the
safety of fish (tilapia) atelocollagen for use in clinical application. We performed in vitro and in
vivo biological studies of medical materials to investigate the safety of fish collagen. The extract
of fish collagen gel was examined to clarify its sterility. All present sterility tests concerning
bacteria and viruses (including endotoxin) yielded negative results, and all evaluations of cell
toxicity, sensitization, chromosomal aberrations, intracutaneous reactions, acute systemic
toxicity, pyrogenic reactions, and hemolysis were negative according to the criteria of the ISO
and the Ministry of Health, Labour and Welfare of Japan. The present study demonstrated that
atelocollagen prepared from tilapia is a promising biomaterial for use as a scaffold in
regenerative medicine.

Materials Tilapia fish scales (Oreochromis niloticus) were obtained from the local market at Tanjung
Malim, Perak (Malaysia) whereas Alcalase 2.4L (declared activity of 2.4 AU/kg and density of 1.18 g/ml),
an endoproteinase from B. licheniformis, was purchased from Brentag specialities, (USA). Collagen
extraction was prepared by using deionized water. All reagents used were of analytical grade.
Extraction of collagen from Tilapia fish scales Collagen was extracted from Tilapia (O. niloticus) fish
scales according to the method of Zainol et al. using hydrothermal method [24]. The fish scales were
collected and washed several times with deionized water to remove dirt. This process is repeated until
the fish scales were cleaned before being dried. The dried fish scales were ground into powder using
domestic grinder, Panasonic MX-337. One hundred grams of fish scales powder was mixed with 1000 ml
of deionized water and extracted hydrothermally at 80°C fo

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Oreochromic Niloticus (tilapia)
scales as tiles (Science
Investigatory Project)
Chapter 1

INTRODUCTION

Building commercial and residential infrastructures in our country is fast growing. One of the building materials is
ceramic tile that is used as floorings in bathrooms, dining area, function halls, etc. Because of this, there is a demand
of ceramic tiles and its industry is booming.

On the other hand, every year, various solid wastes in our country have been great problem to our government. One
example is the scales of oreochromic niloticus commonly known as tilapia found in markets, fish ports and etc. It
makes the surroundings smells bad and can harm to your health. It also serves as silt for reproduction of flies and
other oil-causing insects, which are carriers of disease-causing bacteria and viruses.
This scales composed of a surface layer containing hydroxyapatite and calcium carbonate, and a deeper layer
composed of mostly collagen. The presence of calcium carbonate would make it an ideal component for tiles.
This information brought the idea to the researchers to use the oreochromic niloticus (tilapia) scales as raw material
for ceramic tile making. Due to its high concentration of calcium carbonate, the proponents therefore would like to
substitute it for the main material in ceramic tile making.
Statement of the Problem
The following problems are tend to be answer:

1. Can oreochromic niloticus (tilapia) scales substitute for silica in ceramic production.
2. Can oreochromic niloticus (tilapia) scales make tiles more durable than the commercial tiles.
Significance of the Study
This study will be conducted to eliminate solid waste pollution caused by oreochromic niloticus (tilapia)
scales in the markets, fish ports and etc. by recycling it. Moreover it can also prevent the rapid growth of
population of insects like mosquitoes living in the scales, which are the carriers of disease-causing
bacteria and viruses. In addition, new product means new opportunity for export and new hope for
economic progress.
Scope and Limitation
The focus of the study is on the utilization of oreochromic niloticus (tilapia) scales as raw material for
ceramic tiles. The process of ceramic tile making including tests on properties such as impact strength
and porosity are therefore incorporated in the study.
Definition of Terms
Ceramic tile – is the tile the will be made from oreochromic niloticus (tilapia) scales and some basic
components of a commercially available ceramic tile.
Impact Strength – is the ability of ceramic material to bear crushing loads.

Tilapia Scales – are the scales derived from oreochromic niloticus.

Porosity – is the penetration of liquids and vapors through the material that usuallycauses structural
damage.
Acetone is a good solvent for many plastics and some synthetic fibers. It is used for
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superglue before they harden. It is used as one of the volatile components of some
paints and varnishes.
Acetone - Wikipedia
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Acetone (CH 3COCH 3), also called 2-propanone or dimethyl ketone, organic solvent
of industrial and chemical significance, the simplest and most important of
thealiphatic (fat-derived) ketones. Pure acetone is a colourless,
somewhat aromatic,flammable, mobile liquid that boils at 56.2 °C (133 °F).
acetone (CH3COCH3) | chemical compound | Britannica.com
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