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RESEARCH METHODOLOGY
employed in the study. Specifically, it aims to discuss the research design that was
chosen for the purpose of the study, the sources of data under analysis, the
instrument used for data collection, the procedure followed to carry out the study,
the statistical treatment used to analyze the gathered data, the breakdown of cost
and expenses and lastly, the blueprint of the product under study.
Research Design
for facemask.
The researchers use the experimental method for the study. Experimental
one or more variables, and controls and measures any change in other variables.
Using control and manipulation on the variables on the experiment aims to see if
the variables if the data that are manipulated has an effect to the results. With this,
inferences from the research. This method ensures that there will be no bias or
any mistakes in the experiment that would otherwise compromise the results of the
Research Instrument
The data in this study were gathered by the use of laboratory experiments.
The extracted collagen sample and the produced face masks are subjected to
Centrifuge machine
Petri dishes
Watch glass
Table1
Raw materials, Tools, Equipments and Chemicals used in the Study
This study aims to analyze the quality of the face mask made out of
mask products. However, before the product is made, collagen is first needed to
be extracted from the fish wastes. With that being said, it will undergo different
kinds of processes in order to attain its objective. After which Extracted collagen
from the market and it will be used for the collagen extraction. A method on the
(2016) study with minor modifications. All undergone processes were carried out
With the use of scissors, the fish skins were cut into small pieces of
0.5 cm in length. The skins were then extracted with the use of 0.1M NaOH
with a sample to alkali ratio of 1:8 (w/v) in order to withdraw the non-
collagenous proteins present in the skin. It was mixed for 6 hours; however,
the alkaline solution was replaced with a new one for every 3 hours. In order
to obtain neutral pH, the next samples were cleansed with distilled water.
to alcohol ratio of 1:10 (w/v) overnight in order to remove the fats from the
fish skin. Again, the samples were cleansed repeatedly with the use of
distilled water until the skins were extracted with a variety of acetic acid
concentrations that is ranging from 0.1 to 1.0M. The remains were filtered
with the use of cotton cloth. Proteins are then retrieved with a procedure
called salting-out which uses NaCl to a final concentration of 2.5M with the
was then dialyzed on a 0.1M acetic acid and distilled water. The acid-
the lyophilization.
Hammering of the fish head is done for the extraction of the bone
collagen. It is washed for 5 hours with the use of distilled water. It is soon
the non-collagenous proteins for 24 hours; the alkali solution was replaced
with a new one 4 times since it should be changed every 6 hours. In order
to achieve a neutral pH value, the remains were washed out with distilled
water.
Removing of calcium were done with the bones with the use of 0.6M
butyl alcohol at a sample to alcohol ration of 1:10 (w/v) overnight. With the
use of distilled water, the bones were washed thoroughly; prior extraction
with a variety of acetic acid concentrations ranging from 0.1 to 1.0M for one
then salted out with a NaCl to a final concentration of 2.5M in the presence
acid and those dissolved proteins were dialysed against o.1M acetic acid
and distilled water. A freeze-drying process will then occur for the extracted
collagen content.
hours at a sample to alkali solution ratio 1:8 (m/v) in order to remove the
washed with distilled water. Then, the residue was extracted with a variety
ratio of 1:2.5 (w/v) for duration of one to five days according to the
experiment planned for optimization. Moreover, with the use of cotton cloth,
the extracted collagen from fish wastes will be acquired and the final
acetic acid, and dialyzed against 0.1 M acetic acid and distilled water,
respectively. Lastly, this acid-soluble collagen from fish scales was then
lyophilized.
In extracting the collagen content from the fish fins, the fins will be
washed with distilled water and will undergo lyophilized. The portion that
were insoluble was then extracted with various concentration of acetic acid
ranging from 0.1 to 1.0 M at sample to acid ratio 1:2:5 (w/v) for one to five
days. Then, the extract will undergo centrifugation for 1 hour at 20,000xg to
The first portion which is acid-soluble were salted out with NaCl to a
final concentration of 2.5 M (at neutral pH) in 0.05 M Tris-HCl (pH 7.5).
Afterwards, the proteins that were precipitated was then centrifuged again
at 20,000xg for 1 hr, and then dissolved in 0.5 M acetic acid before dialyzed
against 0.1M acetic acid. After it was lyophilized for the final process, acid-
water and decalcified with 0.5 M EDTA (pH 7.4) for 5 days. The EDTA
solution was changed once a day. With the use of 10 % butyl alcohol, the
decalcified bones were washed for 1 day. Afterwards, centrifugation at
20,000xg was used to remove the extracted collagen and then washed with
distilled water and lyophilized. Lastly, it was dissolved in 0.5 M acetic acid
and dialyzed against 0.1 M acetic acid. As a result, extracted collagen were
The yield of collagen extracted from the fish wastes are calculated
collagen based on the freeze dried weight of the raw materials offish
wastes. The figure below shows the formula on calculating the collagen
yield.
procedure:
1. Unscrew and remove the storage bottle from the electrode (be careful not
3. Rinse the electrode tip with distilled water. If you see bubbles in the
down a thermometer).
4. Start data collection. Place the tip of the electrode in the solution to be
solution.
preparing dry collagen (100 - 200 mg) that is collected from skins, scales,
bones or fins of fish waste that were mixed with 6 N HCl to a ratio of 1:10
(w/v) at 110 °C for 24 hours. The hydrolysate was then added with 2.5 mM
of internal standard and distilled water. The mixture was then refined
Fluor (Waters, USA) reagent and 70 µl of borate buffer. The sample was
then left to stand for 1 minute at room temperature (20 – 25 °C and 5 µl was
loaded to a AccQ Tag Column (3.9 x150 mm) in the HPLC system (Waters,
amino acids in the sample was estimated based on the area under the
concentration of each amino acid as the internal standard method was used
to improve the precision and accuracy of the results where volume errors
are difficult to predict and control. Amino acid composition revealed from
test the odor of the extracted collagen the researchers used the Sniff test.
The results of this test are very dependent to the observers since the
D. Product Making
Sheet masks are face-shaped masks that are usually soaked in serum and
component. In this study, the extracted collagen from fish wastes is used. 1
of natural essences to make a serum. Cinnamon oil (1/4 tsp.) is added for
fragrance and additional nourishment. Compressed masks are soaked into the
serum mixture. Sheet masks are readily available in the market formed into
compressed pellets and once soaked will puff into face-shaped masks. The
mixture is left for 5 minutes. The quality of the masks is then observed and
masks.
Statistical Treatment
The statistical tools used to treat the data of this research are the following:
measure of percent change, which is the extent to which a variable gains or loses
intensity, magnitude, or value. The figures are arrived at by comparing the initial
(or before) and the final (or after) quantities according to a specific formula.
One-tailed z Test. It indicates that the Ho should be rejected when the test
value is in the critical region on one side of the mean. A one tailed z test is either
The figure above shows the prospected image of the research product. The
face mask is generally white in color and is soaked in a serum with the extracted
collagen. The packaging of the product features a jumping fish (which is Tilapia)
out of water in a bluish white background. The product name Nyl is a word play of
the word Nile which is the variety of Tilapia utilized for collagen extraction in this
study.