Abstract especially skin’s damage. Snakehead extract contains abundant

Abstract :

 

   The
extraction of snakehead fish Albumin was conducted by using water, HCl 0,1M,
and ethanol 50% solvent with and without heating at temperature of 50-60?C for
10 minutes. The drying was conducted by mechanic dryer at temperature of 60-70?C.
The parameters of this research were albumin, total protein content, and yield
percentages . The result shows that the fish protein concentrate of snakehead
fish with highest albumin content which is 20,80% while lowest fat 1.78% was
produced by the treatment of HCl 0,1M solvent by heating at temperature of
50-60?C for 10 minutes. Whereas the fish protein concentrate of  snakehead fish with highest yield 6.41% and
total protein content 76.13% was produced by the treatment of water solvent
without heating. The optimal extraction method in extracting the fish protein
concentrate of snakehead fish was the extraction by using HCl 0,1M solvent with
heating at temperature of 50-60?C for 10  minutes. Award cork fish meat or protein
extracts have been tried to increase the levels of albumin in the blood and
help cure some diseases especially skin’s damage. Snakehead extract contains
abundant  albumin content about 2.17 ±
0.14g/100mL which is the largest fraction 64.61% of protein compared to other
fishes . This protein content is sufficient enough to provide albumin for
highly demanded diseases such as hypoalbuminemia,post-surgical patients, and
growing children.

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Keywords :  snakehead extract; wound healing; cure
diseases; Fish protein concentrate, Snakehead fish.

 

 

 

 

 

 

 

 

TABLE
OF CONTENT

 

CONTENT                                                                                   
            PAGE

Abstract                                                                                           
                                       1

Table of contents                                                                                                                    2

CHAPTER 1 :
INTRODUCTION                                                 
                                      3-4

1.1   Background

1.2   Problem
statement

1.3   Aim

1.4   Objectives

CHAPTER 2 : LITERATURE
VIEW                                                                     5-8

2.1
Background

2.1.1 Research background

2.1.2 Morphology

2.1.3 Economic and medical importance

2.2
Wound healing

2.2.1Composition of FPC

2.2.2 Arachidonic acid

2.2.3 Concentrated fish protein aerosol

2.2.4 Wound healing process

CHAPTER 3 : METHOD AND
PROCEDURE                                                       9-11

3.1 Equipment and materials

3.2 Research procedures

3.3 Sample analysing

3.4 Sample preparation

3.5 Protein content

3.6 Yield

3.7 Data analysis

3.8 Gantt chart

CONCLUSIONS
                                                                                                                12

REFERENCES                                                                                                                    13

 

CHAPTER 1

INTRODUCTION

1.1  Background

  Snakehead fish is one of the biological
resources with high economic value due to its high content of albumin (Asfar,
et al., 2007; Tawali, et al., 2012; Mustafa, et al., 2012; Sulistiyati, 2010).
Albumin is a kind of globular protein that dissolve in water, salt solvent and
diluted acid (Winarno, 2004). This fact indicates that albumin protein of
snakehead fish can be extracted to be a concentrated fish protein that can
provide us with multiple usage .

  Fish
protein concentrate is one of product result of fishery processing such as fish
where the protein is a special component compared to other component in fish
body. Fish protein concentrate is sometimes abbreviated with FPC. Fish protein
concentrate is almost similar with fish powder, nevertheless the making of fish
powder is not purposed for human consumption but also for livestock woof
(Windsor, 2001). Fish protein concentrate is one way in providing the fish for
human consumption with protein as the special component.

  The discovering of albumin protein in Snakehead
fish which is beneficial for health will make this fish to have a high
functional potency. The albumin extraction of Snakehead fish for producing the
albumin protein concentrate is expected to be an alternative for cheaper
albumin source for clinical usage. Based on this information, this study was
conducted to investigate the extraction method of Snakehead fish (Channa striatus) into fish protein
concentrate that contains high albumin content.(Fpc et al., 2015)

1.2
Problem Statement

The purpose of this
research is that we want to extract Albumin protein from Snakehead Fish Channa Striatus to produce Fish Protein
Concentrate (FPC) and to study its effectiveness in wound healing properties . Protein is one of the important nutrients  for human in their daily life as it is
required by human’s body such as wound healing that involves skin damage. However,
nowadays albumin protein in the Snakehead fish are not overally used because
there is no furher studies that have been done by other researchers. Besides,
people also lack of knowledges regarding the benefits of Fish Protein
Concentrate as they are suspicious about the quality of the products.  

 

1.3 Aim :

 To extract Albumin protein of Snakehead Fish Channa Striatus to produce Fish Protein
Concentrate (FPC) and also to identify how far its effectiveness in wound
healing properties.

 

1.4  Objectives :

1) To gain information regarding suitable method to
extract Albumin from Snakehead fish

2) To portray accurately the characteristics of a
particular fish ,environment and its additional functions

3) To determine the relative functions of the
Snakehead fish in pain healing, skin’s anti-inflamantory and-microbial agents

4) To identify the effectiveness of Fish Protein
Concentrate (FPC) properties in wound healing 

 

 

 

 

 

 

 

 

 

 

 

CHAPTER 2

LITERATURE REVIEW

2.1
Background

2.1.1 Research background

  
Snakehead fish can be extracted by various methods but for this project,
we are trying to determine the best methods that can be used to extract Albumin
from Snakehead fish. The question that I want to highlight here is that C. Striata  extracts can be used as alternative sources
of albumin protein and is it effective in wound healing for humans . This
question has sparked debate between different authors becouse the fields that
they are studying may be different with each other. One of the best extraction
method that can be used to extract albumin is by using HCl 0.1M solvent with heating
at temperature of 50-60?C for 10 minutes. The results shows that fish protein
concentrate of Snakehead fish contains highest albumin content and lowest fat
level which is 20.80% and 1.78% respectively . According to the authors (Abu Bakar
Tawali, Nurlailah Abdullah, Meta Mahendradatta , 2014 ) ,they quoted these
results in the journal tittled “Extraction of Albumin of Snakehead fish (Channa
Striatus) in producing  Fish Protein Concentrate
(FPC)”. They concludes that Snakehead fish albumin protein concentrate that contains
highest albumin 20.80% with lowest fat content 1.78% is the results from
treatment of HCl 0.1M by heating at temperature of 50-60?C for 10 minutes, whereas
the concentrate with highest yield 6.41% and raw protein content 76.13% comes
from the treatment of water solvent without heating. One of the  succesful extraction method that can be used
to produce Snakehead fish protein concentrate is from HCl 0.1M treatment by
heating the samples at temperature 50-60?C for 10 minutes. As my research
suggest , the extraction of albumin from Snakehead fish can be very useful
especially in curing wounded parts of human’s skin . These authors have
different opinions in their journal regarding this issue and all of authors believed
that their research are more accurate as the samples, methods and procedures
are likely following the actual research as conducted by an expertise
scientists that have authority in their research.   

 

 

2.1.2 Morphology

    Family Channidae have some distinguish
feature among the species. Their head surfaces have large plate-like scales. According
to (Norainy, 2007) , their mouths are large with sharp teeth while their fins
are hyaline without spines. Next, the dorsal and anal fins are also long in
their size .

2.1.3 Economic and
Medical Importance

   Channa striata is widely consumed throughout
the nation not just as food, but also as a remedy for wound healing. Midwives
have constantly promoted C. striata for wound healing despite without realizing
the actual scientific basis to the claimed. Thailand, Indonesia, Indochina,
Philippines, China, and India had extensive cultures of this and it is among
the most popular table fish even in seafood restaurants. Many people believe
that C. Striata contains all essential elements to bring about good health and
helps to recover the lost energy after long illness (Mat Jais et al., 1994,
1997, 1998). (Ng and Lim ,1990) and ( Lee and Ng, 1991) indicate that Channa
striata, along with other fish types  are
utilized for medicinal purposes; particularly in Indonesia and Malaysia. Cream
extracts of Haruan tissues also contain high levels of arachidonic acid which
is a precursor of prostaglandin, essential amino acids and polyunsaturated
fatty acids that are necessary to promote prostaglandin synthesis. Treating
wounds with these extracts has been demonstrated to promote synthesis of
collagen fibers better than standard use of cetrimide, an antimicrobial quaternary
ammonium compound, thus increasing tensile strength (Baie and Sheikh, 2000).

 

2.2
Wound healing

2.2.1 Composition of
Fish Protein Concentrate (FPC )  

Wound healing is a
complex phenomenon that results in the treatment of anatomic continuity and
function, accomplished by several processes involving the orderly sequence of
biological events such as inflammation, cell migration, angiogenesis,
provisional matrix synthesis, collagen deposition and re-epithelization. The
wound healing properties of C.Striatus are attributed to its fatty acid and
amino acid composition. C.Striatus or Haruan’s mucus and tissue extracts are
found to contain high amount of amino acids especially glycine and arachidonic
acid. These two materials are reported to promote wound healing by initiating
collagen synthesis and re-epithelialisation in damaged tissues. Hence, haruan
extracts are recommended for post operative wound healing as well as post
pregnancy rehabilitation. Haruan is also known to produce polyunsaturated fatty
acids which can regulate prostaglandin synthesis that enhance wound healing
properties .   

2.2.2 Arachidonic acid

Haruan contains
arachidonic acid (C20:4) as 19.0%, which is a precursor for prostaglandin and
thromboxane biosyntheses interfering with blood clotting process and its
attachment to endothelial cells promoting wound healing. Both fatty and amino
acids are part of important components for wound healing processes. Hence,
deficiency in these essential components will hinder the recovery process.
Glycine which is a major component of human skin collagen, together with other
essential amino acids such as alanine, proline, arginine, serine, isoleucine
and phenylalanine can form a polypeptide promoting re-growth and tissue healing.

2.2.3 Concentrated fish
protein aerosol 

Traditionally, wound
treatment has been carried out by letting the wound dry to create a scab by
itself. For the past three decades, wound treatment has undergone changes and
new techniques have been introduced. One among them includes the use of
artificial wound dressing to cover the wound to maintain high humidity level
thus stimulating the epidermis to reproduce fast. Though a number of spray
products or aerosols for wound dressings are available in the market, (Febriyenti
et al.) have formulated an aerosol concentrate containing a mixture of Haruan
extract and a film-forming polymer. When the fish protein concentrate is
sprayed on the wound,it will formed a thin layer of dressing and the added
haruan extract will automatically enhanced the healing process.

2.2.4 Wound healing
process  

 Haruan treatment promoted remodeling of collagen,
by the synthesis of inter and intra-molecular protein cross linking and thus
produced a marked increase in tensile strength as compared to the cetrimide
treated group Gen Pharmacol. 1994 . Various studies
have shown positive effects on patients’ recovery during post-operative
duration  (Jais et al., 1994; Wee, 1982).
According to Baie and Sheikh ,2000 , application of cetrimide cream made with
snakehead extracts on wounds resulted in greater tensile strength on wounds.
These effects may have been attributed through the impeccably high levels of
wound healing amino acids namely aspartic acid, glutamic acid and glycine
(Chyun et al. 1984). A key component of human skin collagen, glycine interacts
with aspartic acid and glutamic acid to generate polypeptides are needed for growth
and healing (Chyun et al. 1984; Heinmann, 1982). High levels of essential fatty
acid such as arachidonic acid, 20:4?6 also present in Fish Protein Concentrate (Jais
et al., 1994; Zuraini et al., 2006). Arachidonic acid is the most important prostaglandins
precursor which induces platelet aggregation and endothelial cell adhesion
which in turn initiates blood clotting (Bowman and Rand, 1980) while stimulating
blood cell responses to injury. This species is known for having the beneficial
aspects of anti-inflammatory, antimicrobial and anti-cancer properties .
Studies have also shown that C. striatus has a good antinociceptive property
that makes it suitable for reduction of post-operative pain in a manner
comparable to morphine .

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

CHAPTER 3

3.0  MATERIALS AND METHODS

3.1. Equipment and
Materials

   Materials used in this study were Snakehead
fish (Channa striatus) obtained from nearest wet market in Nilai , Negeri
Sembilan. All chemicals for protein analyzing (Lowry method)  Na2CO3, NaOH, CuSO45H2, NaK-tartrat, Bovinalbumin
Folin-Ciocalteu Phenolreagenz, aluminium foil, roll tissue, plastic clip, plain
plastic, and labeling paper can be prepared in laboratory of Faculty Science
and Technology. All chemical materials for Kjehdahl method such as concentrated
sulfuric acid, oxide hydrargyrum, sulfuric potassium, hydroxide-sodium
thiosulphate solution, saturated boric acid solution, chloride acid solution,
and diethyl ether solvent are purchased from the overseas suppliers such as
Sigma Aldrich which originated from St.Louis, Missouri, United States .

3.2 Research Procedures

  Extraction of albumin protein where Snakehead
fish was weeded (process of removing  scales, gills, and stomach content) .Then ,we will
washed until there is no more blood and mucus. First, it must be cutted to
smaller pieces and  the bones must be
removed. Then it was smoothed using blender by adding solvent with ratio 1:1
(100ml solvent : 100g fish). Treatments for this research were as follows, A1B1
which is water solvent without heating while 
A1B2 is water solvent with heating at temperature
50-60?C for 10 minutes; A2B1 which is HCl 0.1M solvent
without heating while A2B2 is HCl 0.1M solvent with
heating at temperature 50-60?C for 10 minutes; A3B1 which
is 50% ethanol solvent without heating while A3B2 is
ethanol 50% solvent with heating at temperature 50-60?C for 10 minutes. Sample
from each treatments will be filtered to separate liquid and dregs. Liquid must
be separated from its oil by adding 200ml of hexane solvent then shaken for 30
minutes. After forming two different layer, the oil will be separated by
funnel. Extracted liquid was dried in oven at temperature of 60-70?C. The dry
extract need to be measured and analyzed for specific purposes.

 

 

 

3.3  Samples Analyzing

      Dry albumin protein extract will be
analyzed for water, albumin protein, total protein, total fat, and yield
content (Apriyantono, et al., 1989). Smoothed materials will be weighed as many
as 3 grams then covered with aluminum foil that has been weighed .Materials are
dried in oven at temperature of 100- 105?C for 3-5 hours, then chilled in
desiccators and weighed. Materials then was dried again in oven for 30 minutes,
chilled in desiccators and weighed. This trial must be repeated untill the
weight become constant. Calculation for water content used this formula:

 x 100%

 

3.4 Sample Preparation

     Samples must be in liquid state by adding some
water. The solid phase will be filtered after centrifugation. Attention should
be given to dilution factor in determining samples where 0.1–1 ml of sample is
pipetted and put into test tube, then treated in the same way to standard
determination  .          

3.5 Protein Content
(Kjeldahl-Micro Method)

     Approximately 0.5 gram of sample is
weighed carefully, then added into kjedahl flask 100 ml. After that, an approximately
1 gram mixture of Selenium and 10 ml concentrated H2SO4
are added. Khjedhal flask with its content need to be shaken untill all the
sample wetted by H2SO4 . After that, it is destructed in
acid cupboard untill clear. The solution is left to cool then quickly poured into
volumetric flask 100ml and rinsed by aquades, then added aquades untill sign.
An Erlemeyer consisted of 2% , 10 ml of H3BO3  and 4 drops of mixture indicator solution in
Erlemeyer 100 ml is prepared. 5 ml NaOH 30 % and 100 ml aquades are pipetted,
then distilled untill the container is filled about 50 ml. The end of distiller
is rinsed using aquades then the container and its content is titrated using
HCl or  H2SO4 with
0.0222 N solution untill the colour changed into light red and does not
disappear for 30 minutes.

 

 

3.6 Yield

  To calculate the yield of albumin protein
concentrate, the formulation used was as follow :

 Yield (%) =  

3.7  Data Analysis

   Data
analysis was conducted using data processing method of quantitative descriptive
and Complete Randomized Design with four fold replication.

 

Gantt
Chart Planning  2017/2018    :

 

 

 
 
December

 
 
January

 
 
February

 
 
March

 
 
April

Research

 

 

 

 

 

Initial
report

 

                       

 

 

 

Cording
& GUI

 

 

 

 

 

Final
report

 

 

 

 

 

 

 

CONCLUSION

 

   As
conclusion, Haruan is a freshwater medicinal fish inhabitant in several Asian
countries and still be used as medication to treat wounds, alleviate pain and
boost energy. Research findings have put a light on its traditional uses by
elucidating possible compounds that may give rise to the observable therapeutic
effects. Snakehead fish extract may also have a role in other non-traditional
uses such as in treating neurological diseases and in inducing regenerative
potential of organs and cells. Future work on this therapeutical fish must be
continued and not only limited to its previously known, traditional uses. Overally
,for Fish Protein Concentrate there are three types of protein powders which
were type A,type B and type C. For type A, it is virtually odourless and
tasteless powder that having a maximum total fat content about 0.75%. For type
B, it is a powder that having maximum fat content of 3%. Last but not least is
type C which is from Snakehead fish and also known as meal that was produced
under satisfactory hygienic conditions.To date, only liquid Snakehead essences  are used for consumption purposes . Creams
that are used for external application on the epidermal layer are also
available commercially in the market. These products possess many challenges
such as limited shelf life and difficulty in transportation due to bulkiness of
liquid contained within the product.Therefore, if the protein extract
manipulated into dry powder form, the shelf life of Snakehead protein can be
extended due to removal of water and subsequently reduce the bulkiness.
Currently, only few researchers have successfully prepared powder form of
protein extracted from the flesh of snakehead( Hui et al. , 2010) reportedly
prepared K-carrageenan encapsulated snakehead extracts by utilising a spray
dryer while most other researcher prepared liquid extracts by using the traditional
preparation method of cooking snakehead flesh in a pressure cooker (Jais et al.
1994, Dahlan-Daud et al., 2010)

 

 

 

 

 

REFERENCES

 

1) Andrie, M. 2015. Formulasi Salep Ekstrak Ikan Gabus (Channa striata)
dengan Variasi Konsentrasi Basis. Penelitian Dosen Dana DIPA, Pontianak,
Universitas Tanjungpura.

2) Ansel, H.C. 1989. Pengantar Bentuk Sediaan Farmasi. 4th ed (Penterjemah:
Farida Ibrahim), UI Press, Jakarta, pp. 490-494.

3) Anton, T.J. 1989. Occupational Safety and Health Management, McGraw-Hill
Book.Co, Singapore, pp. 118-121.

4) Carville, K. 2012. Wound Care Manual. 6th ed, Silver Chain Foundation,
Western Australia.

5) Collins, N., and Sulewski, C. 2011. Ostomy Wound Management, pp. 10-13.

6) Gadekar, R., Saurabh, M.K., Thakur, G.S., dan Saurabh, A. 2012. Studi of
Formulation, Characterisation and Wound Healing Potential of Transdermal
Patches of Curcumin. Asian J Pharm Clin Res, 5:225-230.

7) Gusdi, O. 2012. Formulasi sediaan gel ekstrak ikan gabus (Channa
Striata) sebagai obat luka sayat, Shafri, M.A., dan Mat, J.A.M. 2012.
Therapeutic Potential of Haruan (Channa striatus): from food to medicinal uses,
Mal. J. Nutr, 18:125-136.

 8) Tarigan, R., Pemila, U. 2007.
Perawatan Luka: Moist Wound Healing. Makalah, Jakarta, Universitas Indonesia.

 9) Asfar, M., A.B. Tawali, Pirman,
2007. The Extraction of Albumin of a Snakehead (Channa striatus) as a food
suplement. Thesis. Faculty of Agriculture Hasanuddin University (in
Indonesia)

10) Mat Jais, A.M., McCulloh, R., Croft, K., 1994. Fatty acid and amino
acid composition in haruan as a potential role in wound healing. General Pharmacology
25, 947–950.