RATIFICATION PAGE

Complete Report of Biotechnology Experiment with the title “Virgin

Cocconut Oil”which made by:
name : Fatmah Kamaruddin
ID : 1614440002
class : Biology Of Education ICP 2016
group : I (One)
After checked and approved by assistant and assistant coordinator, this report was
accepted.

Makassar, April 2019

Assistant Coordinator, Assistant,

Suharyanti Amir , S.Pd Eka Wahyuni

ID. 1314141011

Known by,
Responsibility Lecture

Prof. Dr. Ir. Hj. Yusminah Hala, M.S.

NIP: 19811212 198601 2 002
 CHAPTER I
INTRODUCTION

A. Background

Virgin Coconut Oil or pure coconut oil is made from fresh coconut meat. The
process is all carried out in relatively low temperatures. Squeezed fruit juice of
coconut milk. This coconut milk is further processed through relatively low
temperature heating, fermentation, cooling, enzyme addition, mechanical pressure
or centrifugation. The addition of inorganic chemicals and chemical solvents is
not used and the use of excessive high temperature is also not applied. The result
is pure coconut oil which tastes soft and has a unique coconut smell. When frozen
the color is pure white and in a liquid state that is colorless or clear.
Virgin coconut oil (VCO) is growing in popularity as functional food oil and
the public awareness of it is increasing. It is expected that VCO will experience a
dramatic growth in the market. The introduction of VCO has open up new
research that basically reveals new things besides what has already been known
on commercial coconut oil. This paper mainly discusses on some of the findings
associated with VCO up to date. Physicochemical properties, antioxidant activity,
clinical and authentication. Coconut oil is extensively used for food and industrial
purposes. The oil is rich in medium chain fatty acids (MCFA) and exhibits good
digestibil. Various methods have been developed to extract coconut oil, either
through dry or wet processing.
Dry processing is the most widely used form of extraction. Clean, ground and
steamed copra is pressed by wedge press, screw press or hydraulic press to obtain
coconut oil, which then goes through the refining, bleaching, and deodorizing
(RBD) processes. During the RBD process, heating process is applied especially
during deodorization process, which is carried out at high temperature between
204 and 245 . The copra industry also faced some problems such as contamination
by aflatoxin in copra and cake and presence of high free fatty acids due to high
moisture content. Recently, there is a trend towards producing coconut oil which
does not have to go through the RBD process. Rather than going to the normal dry
process, this oil is obtained by wet processing which entails the extraction of the
cream from the fresh coconut milk and consequently breaking the cream
emulsion. This process is more desirable as no chemical or high heat treatment is
imposed on the oil. The coconut oil produced through the wet method is known as
virgin coconut oil (VCO). The term VCO refers to an oil that is obtained from
fresh, mature kernel of the coconut by mechanical or natural means, with or
without the use of heat and without undergoing chemical refining. Unlike RBD
coconut oil which is tailor-made for cooking purposes, VCO is marketed lately as
functional oil. Since its first introduction, virgin coconut oil has captured the
attention of vast majority of publics. VCO is produced through wet method, via
coconut milk. Through the wet process itself, many ways of producing coconut oil
have been described. No specific requirement for producing VCO has been
established. Based on the definition of virgin oil, it is understood that as long as
the oil does not go through the RBD process.
Virgin coconut oil (VCO) directly extracted from fresh coconut meat at 50 C
temperature was tested for its effect on the activities of antioxidant enzymes and
lipid peroxidation levels in male Spragu Dawley rats, compared to copra oil (CO)
and groundnut oil (GO) as control. Oils were fed to rats for 45 days along with a
semi-synthetic diet and after the experimental period various biochemical
parameters were done. Individual fatty acid analyses of VCO and CO were done
using gas chromatography. Effect of polyphenol fraction isolated from the oils
was also tested for the ability to prevent in vitro microsomal lipid peroxidation
induced by FeSO4. The results showed that GO, rich in polyunsaturated fatty
acids, reduced the levels of antioxidant enzymes and increased lipid peroxidation,
indicated by the very high MDA and conjugate diene content in the tissues. PF
fraction from VCO was found to have more inhibitory effect on microsomal lipid
peroxidation compared to that from the other two oils. VCO with more
unsaponifiable components viz. vitamin E and polyphenols than CO exhibited
increased levels of antioxidant enzymes and prevented the peroxidation of lipids
in both in vitro and in vivo conditions.
B. Purpose
To know what is the meaning of vco and then know how the product can get
the hight quality.
C. Benefit
After do the experiment we can know the meaning of VCO and then can know
how to make a product with high quality.
 CHAPTER II
LITERATURE REVIEW

Different from cooking oils which contain long-chain fatty acids, virgin
coconut oil (VCO) has high medium-chain fatty acids, making it a potential
functional food which can provide some health benefits. In this study, our
objective is to investigate the physicochemical properties, antioxidant capacity,
and metal contents of the VCO extracted through four different processing
methods: chilling and centrifugation; fermentation; direct micro expelling-oven
dried; and direct micro expelling-sun-dried processes. We found that the
physicochemical properties, including moisture content, refractive index,
viscosity, iodine value, saponification value, peroxide value, free fatty acid, and
fatty acid content, of all the VCO conform to the Asian and Pacific Coconut
Community (APCC) standard. All of the VCO predominantly contains lauric acid
which is in the range of 48.40%–52.84% of the fatty acid content
(Ghani, et al 2018).
Virgin coconut oil (VCO) is an edible oil obtained from the milk of fresh
and matured kernel of the coconut (Cocos nucifera L.) a tropical plant belonging
to the Arecaceae (palm) family. VCO is colorless with the aroma of fresh coconut,
and it has been largely consumed for many purposes in cooking, bakery,
confectionary, infant foods, and cosmetics. In cosmetics, VCO is utilized as a
substance to enhance beauty, to promote the growth of hairs, and to improve and
moisturize skin. Another emerging application of VCO is in the health supplement
area, due to the health benefits of medium-chain fatty acids (MCFA) contained in
VCO. MCFA such as lauric, myristic, palmitic, capric, stearic, oleic, and linoleic
acids are easily digestible (Ghani, et al 2018).
Virgin Coconut Oil (VCO), is a product that is very popular consumers,
starting around the Year 2002. This is due to the large number information
obtained, through training, print media, electronics and the internet, about various
benefits from VCO for health. Some consumers who consume VCO produced by
CV. Mas Bening Equator (processing uses gradual heating), revealed recovery
from various types of diseases Their testimony from the benefits of VCO in
overcoming several types of diseases, including: narrowing of arteries, canker
sores, cough, runny nose, hepatitis, diabetes, acid tendons, high cholesterol,
cramps, calcification, ulcer, pain in the knee, heart, ringworm, vaginal discharge,
hemorrhoid,tonsils, being overweight and HIV. In line with the benefits of VCO,
there are various processing methods to obtain VCO in high quantity and quality.
Because of the large number of consumers want to enjoy various benefits from
VCO for health, then almost on every province will be found by processors VCO,
both household and household scale farmer group, so the VCO product young is
obtained (Barlina, 2016).
Coconut plants has great potential to be a product useful preparations.
Virgin Coconut Oil (VCO) is a form of processed meat coconut becomes oil, used
as a drug and can be trusted cure various diseases degenerative for example
cancer, high blood pressure, cholesterol, heart and HIV / AIDS. Excellence This
oil is located in high acid saturated fat which is about 90%make this oil the
healthiest oil Various methods have been used to obtain processed coconut oil
starting from the traditional way up tomodern way. Indonesia as one coconut
producers, have interests the high way to get making coconut oil which is
effective, efficient and the results can compete in the market. To damage the fat
emulsion bond on coconut milk using the method enzymatic. Breaking the
emulsion coconut milk can occur in the presence of an enzyme proteolytic. This
enzyme can catalyze protein breakdown reaction with hydrolyzes the peptide
bond into simpler compounds. One enzyme that can be used to break the peptide
bonds in the emulsion coconut milk is the enzyme papain from sap papaya. The
papain enzyme is an enzyme proteolytic which is a catalyst that catalyzes peptide
bonds in proteins become compounds that are simpler like dipeptides and amino
acids. Papain enzyme including protease enzymes, chemical properties of
enzymes protease depends on the type of chemical group contained in the
enzyme. Because papain has a sulfihidril group on location the enzyme papain is
active sulfihidril proteolytic enzyme group (Silaban et al, 2016).
 Virgin coconut oil (VCO) contains a great deal of MCFAs especially
lauric acid (nearly 50%). Tere is a variety of benefts from MCFAs in VCO; for
example, the MCFAs are digested easily and converted into energy in the liver
directly. Tis makes MCFAs do not take part in the synthesis of cholesterol or
deposit fat in body. Moreover, showed that MCFAs in VCO can control
overweight condition for women. On the other hand, antibacterial ability of
MCFAs is one of the remarkable abilities which is concerned by many authors.
For example, Parfene showed that MCFAs in crude coconut oil could inhibit
Yarrowia lipolyti. The experiments demonstrated the antimicrobial activity of
VCO and MCFAs against Clostridium difcil. Terefore, extracting FFAs from
VCO is necessary to apply in the future research (Nguyen et al 2018).
During the past decade, the incidence of opportunistic fungal infections
has steadily increased. Among these diseases candidiasis constitute real public
health problems in the world, this more especially as the prevalence of both
superficial and deep candidiasis has not ceased increasing in recent years . During
their life 70 to 75% of healthy women have at least one episode of vulvovaginal
candidiasis. The situation is still very worrying in Cameroon among HIV- positive
patients, who have 28.3% gastrointestinal candidiasis, 39.7% oesophageal
candidiasis and 79.6% oral candidiasis. The management of candidiasis is based
on the use of antifungals. However, their use is not without side effects.
Moreover, the emergence of new pathogenic strains and the increase in resistance
to these antifungals increases the risk of infection and predisposes patients to life-
threatening relapses. It is therefore necessary to research and develop new
therapeutic approaches that can be used as an alternative to conventional
therapies. One strategy is the use of white palm kernel oil and virgin coconut oil
used in traditional medicine and potential sources of bioactive compounds (lauric
acid, caprylic acid, capric acid). The objective of this work is to evaluate the
antifungal activity of virgin coconut oil and white palm kernel oil on Candida
species (Kamga, et al 2019).
Consumption of coconut oil has increased over the years. In 2015,
between January and November, exports from the Philippines increased by 61%.1
Virgin coconut oil (VCO) has garnered public and scientific attention as a
potential ‘‘superfood’’ due to its high phytochemical content, particularly of
phenolic compoundsResearch supports the benefit of VCO for a variety of
conditions, including supporting weight loss fighting infection, improving the
cardiovascular disease risk profile (or maintaining its neutrality) and even
lessening the decline in neurodegenerative disorders. Nevertheless, confusion
about VCO continues because it is a highly saturated fat (93%) food. While
researchers have suggested that saturated fat may not be detrimental for health the
most recent 2015 Dietary Guidelines for Americans still suggest limiting saturated
fats, including tropical oils. VCO is a unique saturated fat because it is rich (65%)
in short and medium chain fatty acids that metabolize rapidly without depositing
in arteries and fat cells (Harris et al 2017).
The most substantial medium-chain fatty acid, lauric acid, has
antimicrobial, antifungal, and antiviral properties giving a unique benefit to VCO
compared to other oils. The question remains what impact the resulting 35% long-
chained fats would have on health outcomes. Although the evidence pointing to
the benefits of VCO is increasing, few human studies exist exploring VCO’s
impact on measurable health outcomes. The most recent review on VCO and heart
disease highlights the need for more studies. Most studies that highlighted
beneficial impacts of VCO were in vitro animal studies that used medium-chain
triglycerides (MCTs) or were conducted in Pacific Islander/Asian populations.
The human data in existence are not generalizable to most Western societies due
to different diets, lifestyles, and genetic predispositions (Harris et al 2017).
Microorganisms in food may be harmful in some cases, while in other they
are beneficial. Certai microorganisms are necessary in preparation of foods such
as cheese, pickles, yogurt, and sausage. Other microorganisms, however, are
responsible for serious and sometimes fatal food poisoning and spoilage.
Microbiologists have always been aware that foods, especially milk, have served
as important inanimate vectors in the transmission of disease. Foods contain the
organic nutrients that provide an excellent medium to support the growth and
multiplication of microorganisms under suitable temperatures
(Sherman and Cappucino, 2012).
VCO, canil coconut milk (top layer) is given an inoculum buried a few
days later, the virgin coconut oil (VCO) was collected efficacy as a drug. Medium
for fermentation is usually called a substrate. Usually on technology fermentation
is used as a base material containing carbon. Therefore, mostly from plants and
some from animal products. As an example; grain, milk (milk). Natural raw
material comes from agricultural products and Forest. Carbohydrate; sugar, starch
(flour), cellulose, hemicellulose, and lignin Medium for fermentation are
commonly called substrates. Usually on technology fermentation is used as a base
material containing carbon. Therefore, mostly from plants and some from animal
products. As an example grain, milk (milk). Natural raw material comes from
agricultural products and Forest. Carbohydrate; sugar, starch (flour), cellulose,
hemicellulose, and lignin (Nurcahyo, 2011).
Management of decubitus ulcers using the principle of keeping moisture in
the wound Usually oil-based products are used such as creams, ointments, or oil
products original like VCO. Virgin coconut oil (VCO) is oil produced from
coconut (Cocos nuciferaL.) Fresh and old through a mechanical and natural
process, whether using heating or not this does not affect the transformation of the
oil produced. The effect of VCO on atopic dermatitis shows that VCO has an
effect antibacterial to Staphylococcusaureus colonization in atopic dermatitis
patients.Research on prevention of pressure sores through massage using virgin
coconut oil shows that there are significant differences to the incidence of grade I
pressure sores among respondents who were given preventive care using VCO
with massage and without VCO after being controlled by variable Body Mass
Index (BMI). Virgin Coconut Oil or pure coconut oil is made from fresh coconut
meat. Clinical research and testing has proven the efficacy and efficacy of various
virgin coconut oil diseases and other healthy lives. The mixture of young papaya
fruit skin and leaves can be used as a source of papain for produce enzymatic
VCO. A series of characteristic tests for the resulting VCO has been carried out.
Test the characteristics of physicochemical properties (Setyawati, 2015).
 CHAPTER III
METHODOLOGY EXPERIMENT

A. Place and Date

Day/date : Thursday, April 9th 2019
Time : 09.10-10.50
Place : 2nd Floor Biology Department of Universitas Negeri
Makassar
B. Tools and Materials
1. Tools
a. Basin 2 pieces
b. Filter 1 piece
c. Bottle 1 piece
d. Measure glass 1 piece
e. Statif 1 piece
f. Funnel 1 peace
g. Plastic hose and faucet 1 piece
2. Materials
a. Coconut 10 pieces
b. Alcohol 70% sufficiently
c. Water sufficiently
C. Work Procedure

Mix grated coconut and

enough water to make
coconut milk.

Squeeze and filter the

mixture of grated coconut
and water until it forms
coconut milk.
Installing bottles that have
been assembled with a
hose and faucet at the
stand.

Insert the coconut milk

into a bottle that has been
installed at the stand.

Hollow out the top of the

bottle

Let stand for 2-3 days.

 CHAPTER IV
OBSERVATION RESULT

A. Result

Metode VCO
Observation Result Note
Volume

Acid vinegar 1. Creamy Failed

coconut
milk
2. Water
1

Pineaple 1. Creamy Failed

enzyme coconut
milk
1
2. Water
2

Papaya 1. Oil 29 ml
Leave 2. Creamy
Enzyme coconut
1
milk
3. Water
2
3
Inducement 1. Oil 39 ml
methode 2. Creamy
1 coconut

2 milk
3. Water
3
 Fermentation 1. Oil 87 ml
Methode 2. Creamy
1
coconut
2 milk
3 3. Water
Heating 150 ml
methode

B. Discussion

This time the experiment was conducted with 6 methods in making Virgin
Cocconut Oil. after making the cream on the top layer, then the next step is to
believe the process of making VCO which consists of 6 methods.The acquisition
of VCO by heating method is by heating the coconut cream with a temperature of
80oC to separate Between blondo oil and water, the heating method can shorten
the life of the VCO. The heating method has the best color among the other
methods and the specific gravity of the VCO is in accordance with the quality
standard. Minimum heat is said to be the ideal temperature because at a certain
temperature the water contained in coconut milk will evaporate, thus, the protein
that binds to water will break. Denatured protein will bind the oil from coconut
milk and will also damage it. The coconut oil will be free from emulsion bonds
with protein as its emulgator. With the release of the bond, the oil will collect
separately. The yield produced is relatively small because it is directly filtered by
hand so that a lot of VCO is left or dropped.
In the process this fermentation can use microbes that produce certain
enzymes for break down proteins that bind to oil or carbohydrates so oi can
separate well. The fermentation process by lactic acid bacteria is proven able to
extract oil from coconut milk cream emulsion. During the process fermentation is
the breakdown of coconut cream emulsion compounds. The carbohydrate
compound in coconut milk cream is a carbon source for bacteria Lactic acid is
used as a source of energy in cell metabolism. Formation of lactic acid from
carbohydrates causes a decrease in the pH of the substrate, which can cause that
protein also as an emulsifier in coconut milk cream experiencing denaturation and
clumping. Decomposition of proteins also takes place enzymatically by the
enzyme aminpeptidase which is produced by lactic acid bacteria used as bacteria
starter. Anopopididase is an exopeptidase which describes peptide into amino
acids. With carbohydrate decomposition and protein in coconut cream emulsion,
the oil molecule will be released and forming a separate layer that can be
separated from other parts.
The fishing method will form three layers, where the upper layer is called
cream (oil), the middle layer is blondo (protein) and the bottom layer is water.
Then, the fishing method which has been formed three layers is inserted into a
separating funnel which aims to separate the oil from blondo and water. Oil which
is still slightly mixed with blondo, is carried out by a centrifugation process which
serves to get clear yellow oil separate from all blondo. The feed oil used is good
quality cooking oil. The use of bait oil will greatly affect the yield of oil quality. If
the feed oil used is good quality oil, it will get good oil too, but conversely if the
feed oil used is of poor quality, then the oil yield found will be of poor quality.
Bait oil serves as a breaker or breaks the bonds of fat and protein by binding to
compounds that bind to fat and to draw oils that are still bound to blondo until
they finally unite. Water and oil can unite because there are proteins surrounding
the oil molecules. The fishing method basically transforms the water-oil emulsion
into oils.
VCO (virgin coconut oil) by Enzymatic means the separation of oil in
coconut milk without heating. The bond of oil proteins that are in the emulsion of
ordinary coconut milk is also broken down with the help of enzymes. These are
damaged proteins, not fats. may not change because it will affect the efficacy of
VCO (virgin coconut oil). Therefore, both proportionally and quantitatively, the
fatty acid content may not change. Proteins in lipoprotein bonds are broken down
with protein enzymes, called protease enzymes. In the beginning, enzymes were
found in the body of living things because they help the digestive process in the
body. As time goes on, enzymes can be synthesized or supplied from nature.
Several types of enzymes that can be used to break lipoprotein in the emulsion of
fat are papaya, pineapple, and proteased enzymes originating from river crabs.
Papain enzymes are widely found in papaya leaf sap. While many grumelin
enzymes are found in the hump or liver. Lipoprotein bonds in coconut milk will
also be cut off by themselves.Then, the oil that is bound by the bond will come
out and gather into one. Because oil has a lower mass or specific gravity
compared to water, its position is at the top, followed by protein, and finally or
down, that is water.
Judging from the color, the VCO is the yellow amber originating from the
color of the pineapple pigment, which is anthosanthine which gives a light yellow
to deep yellow color. For the aroma of VCO it smells of sour coconut because
pineapple has an acidic aroma that affects the aroma of the VCO. The rendering is
quite a lot because the enzymatic method produces a lot of yield, an enzyme that
can break down proteins that act as emulsifiers in coconut milk. The breakdown
of coconut milk emulsion can occur in the presence of an enzyme. The length of
the fermentation process also affects a lot of the least yield produced. The longer
the fermentation process, the more VCO produced and the greater the volume of
pineapple and coconut milk extract, the more VCO produced. but in this method 3
layers are not formed.
In the method of acid vinegar, protein destruction or protein denaturation
to be able to obtain coconut oil can be done by acidification. In principle, this
acidification technique is a method of protein denaturation due to the formation of
zwitter ions in electronic ISO conditions. Ion zwiters are formed because
molecules have an opposite charge at each end. In the protein itself it actually
contains NH2 groups which have more posotive charges and negatively charged
carboxylic groups. In order to achieve this electronic ISO condition, the coconut
milk is made in acidic conditions. Usually the pH setting for obtaining an
electrical ISO condition is at pH 4.5 which is done by adding acetic acid
(CH3COOH) or commonly known as food vinegar. With this acidification three
layers will also be formed, where the oil layer is at the top, then the middle layer
of protein and the bottom layer is water, but in this method 3 layers are not
formed. The oil obtained from the color acidification will be clear. but in this lab
it failed to form 3 layers in the vinegar acid method, because there was no
emulsion. this is not in accordance with the theory (Aziz, 2017) that of the main
chemical content in a coconut namely water, protein, and fat. The three
compounds are type of emulsion with protein as the emulgator. Emulsions are
liquids which are formed from a mixture of two or more substances same, where
one substance is inside separate conditions smooth or evenly in other substances.
Meanwhile The emulgator is a substance which is meant function to strengthen
the emulsion. From the protein bond will bind the items coconut oil with a thin
layer so that the oil grains will not be biased join, so does water. Emulsion it will
never break because it's still there is a smaller face tension of water protein from
oil proteins The salting method is carried out with the purpose for solving the
coconut emulsion system by regulating protein solubility inside salt. Protein
contained in coconut milk will dissolve with the addition of salt (salting in), but
under certain conditions protein solubility will decrease along with increased salt
concentration. With a decrease in solubility protein is followed by binding of
water molecules by the salt, which then there is also a separation between liquids
oil with water (salting out). Method making coconut oil by the way salt is done by
adding salt solution in the cream of coconut milk that has been obtained from the
initial stages of making oil. Salt is used as a destroyer of stability emulsion.
 CHAPTER V
CLOSSING

A. Conclussion

VCO is pure coconut oil made from fresh old coconut as a raw material and
processed without heating or with limited heating, highly nutritious and
containing 50-53% lauric acid. VCO is processed by imitating natural methods so
as not to damage the essential content of coconut fat. Making VCO is done by 4
methods, namely heating method. fermentation method, fishing method, papaya
leaf enzyme method, pineapple enzyme method and vinegar acid enzyme method
B. Suggestion

In the next the product must follow work procedures so that the results are
better
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