Formaldehyde Degradation in Tilapia Muscles Using Various Solutions

Amaro, M. D., Dequito, A. M., Galapon, R. G., Malubay, E. Q.
, - Formaldehyde Degradation in Tilapia (Oreochromis niloticus)

Muscles Using Various Solutions
Pacuan, K. A., Ramos, A. C., Villanueva, N. C.
1. INTRODUCTION
1.1 BACKGROUND OF THE STUDY
In consumption of food, besides meat, the other main source of protein that most
consumes, are fish. Fish have always been accessible to every region and has proven to be
one of the most capable of providing a wide range of nutrients. People from all over the globe
have deemed it one of the healthiest foods among every other due to its nutritional value
which contains essentials in fighting micronutrient deficiencies.
Fish, in densely populated countries, is considered to be significant in supplying the
protein intake of residents, given that it is often lower than average. Apart from its health
benefits, fish is often the most easily obtained and affordable food protein in most developing
countries, making it easily the main source of protein of 20% of the world. It is predicted that
the demand for fish will rise to 50% in 15 years’ time, and shipment of fishes from all around
the globe would require perishable food to be delivered.
1.2 REVIEW OF RELATED LITERATURE
The tilapia farming industry in the Philippines have rapidly grew since it was
established in the eighties with the innovation and usage of procedures created by the national
R & D institution regarding the breeding and growing of these fishes in freshwater ponds and
cages. With this continuous growth, this easily made the country the largest grower of
Tilapia, worldwide, having contributed 76,142 metric tons of farmed tilapia. Any household
would tend to ensure that the freshwater fishes they consume are as fresh as they can be. This
makes the marketing channel short such that producers would be able to sell their produce
directly to retailers than having to go to wholesalers, first.
1
Amaro, M. D., Dequito, A. M., Galapon, R. G., Malubay, E. Q., - Formaldehyde Degradation in Tilapia (Oreochromis niloticus)
Formaldehyde is a putrid gaseous chemical used to create building materials, as well
as products used at homes (Beane, 2009). Also mentioned in a study conducted by Hoque,
Raman, and Alam (2018), formaldehyde, when dissolved in water, is called formalin. These
are commonly used as industrial disinfectants and as preservatives in funeral homes and
medical labs. It can also be used as a preservative in some foods and in products such as
antiseptics, medicines, and cosmetics.
Formaldehyde occurs naturally in the environment, but may form as a result of
cooking and smoking. Most living organisms, including humans, make small amounts of
formaldehyde as part of normal metabolic processes (Blair, 2009).
According to the study of Nawshad, Islam, and Khan (2018), formaldehyde can be
harmful to human health. When consumed at a higher concentration, it may cause damage to
internal organs such as the lungs, and may lead to cancer. Formaldehyde exerts burning
sensation and inflammation of the liver and kidneys. In addition, there is evidence linking
formaldehyde with nasopharyngeal cancer because of its carcinogenic properties. Thus, the
use of formaldehyde in food is prohibited in some countries.
Traders and suppliers often resort to using this to preserve the marketability of fishes.
According to Sutapa (2017), recently, several media outlets have reported on formalin
preserved fish across Asia. Fish are dipped in formalin before being transported from fishing
ports to ports, the use of formalin to store fish is widespread. The Food Safety and Standards
Authority of India (FSSAI) Referral Laboratory on fish and fish products and the Central
Institute of Fisheries Technology (CIFT) - Kochi reported the presence of residues of these
chemicals in freshly marketed fish, inter-state fish consignments, and even in ice used during
transportation. Although the amount of formalin in fish decreases overtime during storage, it
2
cannot be removed completely. Consumption of fish adulterated with formalin can cause
health conditions such as abdominal discomfort, vomiting, and renal injury.
Food Safety and Standards Authority of India (FSSAI) officials have also conducted
tests at fish markets and harbors all over India to test for formalin after receiving a lead that
fish sourced from nearby locations were chemically contaminated. Several studies like this
were conducted in India and Bangladesh, giving compact evidence in proving that the use of
formaldehyde in fishes can really be dangerous or can cause serious health problems if not
measured correctly.
As stated by Arashisar, Hisar, and Kaya (2004), there is a specific amount of natural
formaldehyde found in freshwater fishes. The study showed that there is a value of 1.85 µg/g
formaldehyde in Oreochromis niloticus or Tilapia found in ponds of Freshwater Station,
Bangladesh Fisheries Research Institute (BFRI) and Mymensingh.
Although found naturally in food, a fact sheet from the Occupational Safety and
Health Administration (OSHA) states that the risky amount of formaldehyde intake starts
from 0.75ppm and 2ppm as its ceiling. Also, as established by the Agency for Toxic
Substances and Disease Registry (ATSDR), the chronic inhalation minimal risk level of
formaldehyde is set 0.003ppm. Both studies have linked exposure to excessive amounts of
formaldehyde to chronic irritation and in worse cases, cancer.
In an article released by Foodiez News (2011), rinsing the fishes with water is one of the most
effective home remedies in removing formaldehyde content in fishes. In another article, the site
mentioned that adding vinegar to water, creating an aqueous solution, increases the amount of
formaldehyde removed from 60% to 98%. In addition, Bode (2007) mentioned that Turmeric, with
3
Curcumin as its main constituent, serves as a preventive substance against ailments including cancer
that is a risk factor of excessive formaldehyde intake.
Generally, water is an effective way of removing the formaldehyde content in fishes.
In the context of this study, it is used as a component in the solution that serves as a remedy
in the removal of natural formaldehyde occurrence in Tilapia to lessen the health risk factors
of the chemical present.
In getting the accurate level of formaldehyde in Tilapia, the spectrophotometric
method with Nash reagent will be used. In Bangladesh, an experiment was conducted for
analysis of formaldehyde presence in abundant freshwater and marine fish species by
spectrophotometric method. The freshwater fish rohu, Thai koi, and marine fish loyitta,
chhuri sold from local markets were used as caught samples and were tested for detection of
formaldehyde content.
According to Radcliff (2018), the spectrophotometer is a device that is used to
measure light at a specific wavelength. A spectrophotometer consists of two parts, which are
the spectrometer that provides light at a specific wavelength and the photometer that
measures the intensity of light. A research in Syria conducted by Yasri, N., Seddik, H., and
Mossalb M. showed how the spectrophotometric method measured the level of formaldehyde
in natural phenomena including rain and fog. The study developed a selective and simple
spectrophotometric method for the detection of low level formaldehyde, providing accuracy
and precision for the routine application of formaldehyde detection. The developed method
depends on the formation of a red violet color product from the telomerization of FA with TA
in the presence of concentrated sulfuric acid and trace NaNO2 amounts. The colored product
that was formed is stable at a temperature less than or equal to 25 °C and does not interfere
with substances normally present as pollutants.
4
Nash reagent is composed of 3 components which are 2,4-pentanedione (0.2%),
Acetic acid (0.1 M) and Ammonium acetate (3.89 M). A conducted research in North
Carolina states that broken up formaldehyde in aqueous samples was resolved at
submicromolar levels by derivatization with Nash's reagent trailed by fluid chromatography.
Thus, this method requires sample arrangement but even in the presence of other aldehydes
and ketones, the chromatogram is simple.
1.3 STATEMENT OF THE PROBLEM
Formaldehyde is a chemical used for food preservation and pest control. It occurs
naturally in various animals, including fishes, as part of their metabolic process. This has
raised some concerns as the presence of excessive formaldehyde in fishes is known to pose
health risks to humans.
In view of this, the researchers have decided to conduct an experimental study to
determine the amount of formaldehyde present in the tilapia and to know the effectiveness of
three proposed solutions composed of water, white vinegar, and turmeric powder in the
degradation of formaldehyde content in tilapia.
1.4 SIGNIFICANCE OF THE STUDY
This study determined the formaldehyde content of Tilapia (Oreochromis niloticus),
once treated with formalin and cured with the proposed solutions. The researchers used
water, white vinegar, and turmeric powder as removing agents for the formaldehyde content
in Tilapia. This contributes to the society by acquainting consumers with the dangers of
formaldehyde present in food, as it also acts as a chemical to cure fishes from parasites and is
used as a preservative. The study also aims to inform the community of the possible health
risks caused by excessive formaldehyde present in Tilapia. The result of the study could
5
educate the people with the proper and effective removal of formaldehyde that would result
in less possible health risks caused by formaldehyde.
1.5 OBJECTIVES
This study aimed to utilize a solution from locally available materials to degrade the
formaldehyde content in Tilapia (Oreochromis niloticus). The objectives of the study
included the detection of the formaldehyde content in Tilapia. This research aimed to lessen
the formaldehyde content in the fishes. This study also targeted the avoidance of the possible
health risks obtained from excessive amount of the substance. Thus, by formulating an
effective solution, the researchers hope to contribute to the society, consumers, and future
researchers.
1.6 SCOPE AND DELIMITATIONS
The main focus of this research was to find out if the proposed solutions: water; water
and white vinegar; and water, white vinegar and turmeric would remove the formaldehyde
content of the treated Tilapia (Oreochromis niloticus). The research samples that were caught
from Laguna de Bay and bought in Alabang Wet Market consisted of twenty-five (25)
Tilapia: five (5) were untreated and twenty (20) were treated with formalin. This research
was conducted with a limited number of samples and was only done on Tilapia (Oreochromis
niloticus) fish. The primary data gathering method used was done by experimentation that
measured the formaldehyde content and by doing statistical analysis illustrated in the graphs.
The gathered data were analyzed using the limit of detection (LOD) and absorbance.
6
1.7 RESEARCH QUESTIONS
This study aimed to determine the significant changes of formaldehyde levels after the
solutions are utilized as degradation agents. This research aimed to answer the following
questions:
1. How much formaldehyde is present in tilapia based on the results of the
spectrophotometer?
2. How much formaldehyde content is removed in tilapia after the following
treatment?
a. Water
b. Water and Vinegar
c. Water, Vinegar and Turmeric Powder
3. What proportion of the proposed solution aids is the most effective in the removal
of formaldehyde content on a fish?
a. 100% Water
b. 40% Water, 60% Vinegar
c. 40% Water, 50% Vinegar, 10% Turmeric Powder
7
2. METHODOLOGY
2.1 RESEARCH DESIGN
To be able to attain the objectives set forth in this research undertaking, the
researchers employed an experimental study that aimed to detect changes in the levels of
formaldehyde content in Tilapia (Oreochromis niloticus) after using their proposed solutions
that included water, water and white vinegar, water, white vinegar and turmeric powder.
2.2 RESEARCH LOCALE
The collection of data was mostly from the Tilapia fishes from Laguna de Bay to
ensure its freshness. The detection of the presence of Formaldehyde in Tilapia was measured
by the UV-Visible Spectrophotometer that was provided by the Bureau of Fisheries and
Aquatic Resources. The researchers were supervised by one of the authorities in the Bureau
of Fisheries and Aquatic Resources. The experiment required 5% Trichloroacetic acid and
Nash reagent (Acetyl Acetone and Ammonium Acetate) that was also provided by Bureau of
Fisheries and Aquatic Resources. The variables that were required for the solution are mostly
found in markets and these were water, white vinegar, and turmeric powder that were bought
from Alabang Wet Market.
The Tilapia were filleted in one of the researcher’s house to ensure efficient usage of
time for sample preparation. The experimentation was held in the Bureau of Fisheries and
Aquatic Resources because of the availability of the equipment and reagent that the
experiment required, specifically the UV-Visible Spectrophotometer and Nash reagent. The
experiment was conducted on the Aquatic Toxicology laboratory to fulfill its requirement for
a controlled environment.
2.3 MATERIALS
8
The researchers proceeded with an experimental approach in order to gather data for
their study and utilized most of the materials provided by the Bureau of Fisheries and Aquatic
Resources - Aquatic Toxicology Laboratory including the UV-Visible Spectrophotometer
that serves as the main equipment for detecting the amount of formaldehyde in the samples.
Trichloroacetic acid (TCA) was first used in the filleted fish samples in order to break
down the proteins and make it a homogenous mixture with the help of a vortex mixer that
uses a quickly oscillating circular motion.
A centrifuge was used to separate the solids out of liquid chemical solutions of the
samples inside the test tubes by their densities. Then, the liquid solution that remained in the
separation was transferred to a volumetric flask in order to attain a margin of error set at
0.01mL.
Distilled water was then added to the solution to fill up to at least 25mL of the
volumetric flask. Parafilm was also used to cover the flasks that did not have their own
covers for easy mixing.
A pipettor was used to transfer 3mL of the solution to glass test tubes. Nash reagent
composed of acetic acid, ammonium acetate, and acetyl acetone was added to aid the
absorption of light.
A heater set at 60˚C was utilized to submerge the test tubes for the Nash reagent to
react with the formaldehyde content of each sample for 10 minutes. This reagent produces
Diacetyldihydrolutidin complex that is measured to obtain the level of formaldehyde in the
samples.
Five portions of 1mL from each solution was transferred to the sterilized 96-microtiter
plate before placing it on the plate reader for the actual testing of light absorption. The results
9
were then calibrated using a computer program provided by the Bureau of Fishes and Aquatic
Resources.
The visible spectrophotometry method served as an indicator to detect the absorbance
of formaldehyde in the samples. This method was utilized with the application of the
principle of Beer-Lambert’s Law.
10
Fig.1: Vortex Mixer Fig. 2: Centrifuge
Fig. 3: Laboratory heater Fig. 4: UV – Visible

Spectrophotometer
11
25 Tilapia bought
from public market in
Alabang
Freeze for 1 day
5 Tilapia treated with 5 Tilapia treated with 5 Tilapia treated with 5 Tilapia treated with
5 untreated Tilapia 10% Formaldehyde 10% Formaldehyde 10% Formaldehyde 10% Formaldehyd
Use vortex mixer to mix

the sample in tube
Soak in a water and Soak in a water, white
Keep in room Keep in room Soak in water for 1
turmeric solution for vinegar, and turmeric
temperature temperature hour
1 hour solution for 1 hour
Use centrifuge to
separate solid and liquid
components for 10 mins
Fillet, mush, and weigh

5g then put in sample
tube
Treat with 30ml

Trichloroacetic Acid
(TCA)
Transfer the liquid to a

100ml volumetric flask
and mix with distilled
water
Use pipettor to get 3ml of

the liquid sample and
transfer to a test tube
Place in a laboratory
heater in 60 o for 15
mins
Let it cool then shake

once
12
Use the pipettor to

transfer the solution into
the 96 micro plat
13
Put the 96 micro plate in

the UV-Visible
Spectrometer
Use the pipettor to transfer
Amaro, M. D., Dequito, A. M., Galapon, R. G., Malubay, E. the
Q.,solution
- intoFormaldehyde
the 96 Degradation in Tilapia (Oreochromis niloticus)
micro plate
2.5 DATA COLLECTION
The procedure of the data gathering has been done by obtaining information in
libraries and internet research studies that were relevant on the chosen subject of the study
conducted. Prior to the data gathering procedures, letters of permission for the involved
institutes such as Bureau of Fisheries and Aquatic Resources, and Muntinlupa Science High
School were drafted and finalized before being sent either via email or personally presented.
This was done to ensure that there will be no conflict regarding the possible breaching of
research ethics or any legal matters. As soon as the approval from both parties were received,
the researchers began the preparation of the research samples and proposed solutions.
As soon as the 5 set-ups were prepared, (1) untreated with formaldehyde, (2) treated
with formaldehyde, (3) treated with formaldehyde, and soaked in water, (4) treated with
formaldehyde, and soaked in a water, and white vinegar solution, (5) treated with
formaldehyde, and soaked in a water, white vinegar and turmeric powder solution. With the
amount of time indicated in the research design.
Once each set-ups’ samples were completed and placed into their corresponding test
tubes weighing 5 grams each, the samples were taken to the Aquatic Toxicology Laboratory.
There, the actual experiments were conducted. The data were also interpreted into numerical
values to indicate the changes of the levels of Formaldehyde and answer the core research
questions of this study.
2.6 DATA ANALYSIS
14
The data were tabulated and analyzed comparatively. One way of analysis of
Variance was used to assess the significant differences among the different Formaldehyde
content in Tilapia (Oreochromis niloticus) before and after applying the solutions which were
formulated with water, white vinegar, and turmeric Powder. The level of significance was
both set on 0.05 and 0.01. The error presented in the result may be assumed as the standard
deviation and the number of analysis per sample is five (n=5).
3. RESULTS AND DISCUSSION

3.1 RESULTS
The experimentation of the study utilized various solutions such as water, vinegar and
turmeric powder as a removing agent of formaldehyde content in fishes. The study focused on
15
Results
518.37
512.23
495.07
429.37
3.81
U n t r eat ed Tr eat ed Tr eat ed w i t h W at er Tr eat ed w i t h w at ee U n t r eat ed

an d v i n eg ar
determining the amount of formaldehyde content in each of tilapia (Niloticus oreochromis) after
exposing it to the various solutions .
TABLE I. Determining Levels of Formaldehyde Content of Tilapia (Oreochromis niloticus) Using
Various Solution.
Table 1 shows that the result of the light detection in the solution with use of limit of detection
values provided by Aquatic Toxicology Laboratory. For untreated fishes, the limit of detection is at
0.17 µg/g resulting to 3.81±2.13 (µg/g) formaldehyde content. For Treated fishes, the limit of
detection 0.03 µg/g resulting to 518.37±28.84 (µg/g).For the sample treated with Water, The limit of
detection 0.11 µg/g resulting to 495.07±25.37 (µg/g). For the sample treated with water and vinegar
fishes, the limit of detection 0.17 µg/g resulting to 429.37±98.62 and lastly the fishes treated with
Water, Vinegar and Turmeric has a limit of detection 0.11 µg/g resulting 512.23±11.58 (µg/g).
16
Figure 1 Determining Levels of Formaldehyde Content of Tilapia (Oreochromis niloticus)
Using Various Solution.
This graph illustrates the amount of formaldehyde content of Tilapia
(Oreochromis niloticus) in each set-ups. It shows that the highest formaldehyde content is
the “treated” set-up. It has 518.37 µg/g amount of formaldehyde. The second to the
highest amount of formaldehyde is the set up “treated with water, vinegar and turmeric
powder” with an amount of 512.23 µg/g. The third is the set-up “treated with water “it has
a 459.07 µg/g. The fourth is the “treated with water and vinegar” with an amount of
429.37 µg/g. The untreated set-up gained the lowest amount of formaldehyde which is
3.81
Formaldehyde Absorbance Slope 0.02225

(x) (y)
0 0.06893
3 0.1275 Intercept 0.06582
6 0.20073
12 0.33293 r2 0.9999
24 0.60047
30 0.73273
Table 2.1. Calculated Slope, Intercept

Table 2. Standard Calibration Curve
and r2
Table 2 and table 2.1 illustrates the standard calibration curve of the formaldehyde content
and the Absorbance of light in detecting the formaldehyde in UV- spectrophotometer. The
measurements were given by the Bureau of Fisheries and Aquatic Resource- Aquatic Toxicology
Laboratory.
17
Figure 2. A calibration curve
Calibration curve showed linear responses with linearity of coefficient r2, of 0.99. Linearity range is
from 0 µg/g to 33 µg/g Figure 2 shows a calibration curve
Sample Aliquot Average Sample Formaldehyde Sample Average Standard

Codes HCOH absorbance mass amount Derivation
(x’) (y’)
U 4.6822 0.17 5 3.81 0.762
T 1.6099 0.03 5 518.37 115.674
TW 1.9856 0.11 5 495.07 99.014 33.308 5
TWV 4.6822 0.17 5 429.37 85.874
TWVT 1.9856 0.11 5 512.23 102.446
Table 3. Result of Concentration of Formaldehyde Content of Tilapia
(Oreochromis niloticus) Using Various Solution.
This table showed the calculated Aliquot HCOH (x’) and the Average absorbance (y’)
18
of the Tilapia (Oreochromis niloticus) per set-up. It was calculated by using the formula of
Beer Lambert law.
3.2 DISCUSSION
The purpose of this experimental research is to determine which proposed solution aids is the
most effective in the removal of formaldehyde content in Tilapia. This research aims to answer the
following questions:
1. How much formaldehyde is present in Tilapia based on the result of the spectrophotometer?
2. How much formaldehyde content is removed in Tilapia after the following treatments?
a. Water
b. Water and Vinegar
c. Water, Vinegar and Turmeric
3. What proportion is most effective?
a. 100% Water
b. 60% Vinegar, 40% Water
c. 50% Vinegar, 40% Water, 10% Turmeric Powder
The researchers presented their findings in a form of tables and graphs. These data represent
the levels of formaldehyde measured from different set-ups after the experimentation. An
experimental research design was the most appropriate for this research because this design enables
the researchers to test their hypothesis by reaching valid conclusions about the relationship between
the independent variables which are the proposed solutions and the dependent variable which are the
25 samples of tilapia.
19
In measuring the limit absorbance of formaldehyde present in the tilapia based on the results of the
spectrophotometry, the researchers used the limit of detection of each samples and calculated its
mean, and got a result of 0.118µg/g. With the help of the Spectrophotometric method and Nash
Reagent, the researchers found out that the level of formaldehyde present in Tilapia (Oreochromis
niloticus) is 3.81µg/g. It shows that there is a naturally occurring formaldehyde content in the fish
and that the amount present was dangerous for the health with regard to the Occupational Safety and
Health Administration (OSHA)’s statement: “the risky amount intake starts from 0.75ppm and 2ppm
as its ceiling”.
After performing the following treatments with the solutions: water; water and vinegar; and
water, vinegar, and turmeric, the researchers found out that water removed 23.3ppm of
formaldehyde, water and vinegar removed 89.0ppm of formaldehyde, and water, vinegar, and
turmeric removed 6.14ppm of formaldehyde. According to the results, the 60% Vinegar and 40%
Water solution is more effective after it was able to remove 65.7ppm more formaldehyde than the
100% Water in the Tilapia. It shows that Vinegar and Water can be combined in order to lessen the
formaldehyde content of a fish.
CONCLUSION
The conducted study regarding the determination of the levels of formaldehyde contents
found in Tilapia (Oreochromis niloticus), after being treated with the different proposed solutions by
the researchers, came to a conclusion that the most effective removing agent was the water and
vinegar solution.
The amount of formaldehyde detected from the different set-ups has shown that this is
the most effective, due to it having the most difference among others. And though it may
20
have the most significant effect, the formaldehyde content wasn’t completely degraded and
removed as the reading from the spectrophotometer indicated, that the levels from each set-up
were still at a high rate; thus, showing the ineffectiveness of the other materials used in the
study.
Therefore, the researchers proved from the experiment the irreversible effect of
formaldehyde towards food, specifically tilapia. This also resulted to the conclusion that any
surface of food, even with the formaldehyde solution set at a percentage of 1%, it
immediately sets, raising the content at an extreme rate; making thorough removal of the
substance too difficult to be done. Thus, this emphasizes the dangers and consequences of the
chemical substance, once in contact with fishes meant to consume, is to health.
The experiment aided by the light absorbance in the solutions provided an insight on
the possible agents effective for removing the formaldehyde in fishes. This helped the
researchers analyze the results that could potentially help the community in providing a risk-
free environment for food consumption in fishes with the use of simple household materials.
RECOMMENDATION
After conducting the study on the effectiveness of different solutions on removing the
formaldehyde in Tilapia (Oreochromis niloticus), the start of various studies with correlation to
this research may be a new and improved. This study may serve as a foundation for future
studies in the field of aquatics, particularly tilapias and health to lessen the health risks obtained
from marine animals consumed by the Filipino household. The researchers suggest a more in-
depth exploration of the local marine-related issues and studies to avoid creating a gap in local
and foreign literature that serves as the bases for the whole study. The level of formaldehyde
used as part of the control setup and the amount of turmeric powder used in the experimental
setup should also be carefully modified to avoid a disproportionate solution and to prevent
further errors. It is also recommended that future researchers utilize other locally available
21
household materials as removing agents for the formaldehyde content in the fishes like citric
acids, and baking soda. To avoid ethical issues and to abide for legality purposes, it is also
advised to strictly follow and research more on the rules and regulations with regards to the use
of fish as it is under the Vertebrates Animal Section as stated by the Philippine Animal Welfare
Society (PAWS). Furthermore, a larger scale research may be conducted on saltwater fishes or
on a different species of freshwater fishes for a clearer comparison and interpretation of data.
22
REFERENCES
Arashisar, Ş., Hisar, O., Kaya, M., & Yanik, T. (2004). Effects of modified
atmosphere and vacuum packaging on microbiological and chemical
properties of rainbow trout (Oncorynchus mykiss) fillets. International
journal of food microbiology, 97(2), 209-214.
Bianchi, F., Careri, M., Musci, M., & Mangia, A. (2007). Fish and food safety:
Determination of formaldehyde in 12 fish species by SPME extraction and
GC–MS analysis. Food Chemistry, 100(3), 1049-1053.
Jones, S. B., Terry, C. M., Lister, T. E., & Johnson, D. C. (1999). Determination of
submicromolar concentrations of formaldehyde by liquid
chromatography. Analytical Chemistry, 71(18), 4030-4033.
Noordiana, N., Fatimah, A.., and Farhana, Y. (2011). "Formaldehyde content
and quality characteristics of selected fish and seafood from wet
markets." International Food Research Journal 18(1).
Nowshad, F., MdNazibul I., and Mohidus, S. K. (2018).
"Concentration and formationbehavior of naturally occurring
formaldehyde in foods." Agriculture & Food Security 7(1), 17.
Sutapa S, Krishnendu S., Swasti S., Samir B. (2017). Formalin in fish trading: an
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inefficient practice for sustaining fish quality. Arch. Pol. Fish.25: 43-50
DOI 10.1515
Uday N. D., et al. (10 Nov 2018) International Journal of Current Microbiology and
Applied Sciences. Retrieved from https://www.ijcmas.com/7-11-
2018/Uday%20Narayan %20Das,%20et%20al.pdf
Uddin, M., et al. (2014) "Analyzing time dynamic concentration of
formaldehyde in fresh and formalin treated fish ‘Labeorohita’."
International conference on chemical engineering.
Weng, X., et al. (2009). "Rapid detection of formaldehyde concentration in food on
a polydimethylsiloxane (PDMS) microfluidic chip." Food
Chemistry 114.31079-1082.
Yasri, N. G., Seddik, H., & Mosallb, M. A. (2015). Spectrophotometric determination
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24
Appendix A:
PICTURES OF THE STUDY
25
Appendix A
Picture of the Study
Inquiry at Bureau of Fisheries and Test tubes provided by BFAR

Aquatic Resources
Fig. 2.1: Tilapia (Niloticus oreochromis)

Fig. 2.2 : Materials needed in the study
bought from Alabang wet market
Fig. 3: Three solutions (Water, Water Fig. 4: Preparing the samples

and Vinegar, and Water, Vinegar and
Turmeric)
26
27
Fig. 8.2: Using a pipettor, transferring Fig. 9: Putting Nash Reagent to the test
3mL of the solution to the test tubes tubes
Fig. 10: Heating the samples in 60 Fig. 11: Transferring the solution in the
degrees celcius 96 microtiter plate
28
Fig. 11.1: Putting the 96-microtiter Fig. 12: With Sir Marc and Sir James
plate into spectrophotometer to from Bureau of Fisheries and Aquatic
determine the level of formaldehyde in Resources
each samples
29
Appendix B:
DATA TABLES
APPENDIX B
30
Computation and Data Analysis
Sample Test Parameters Test Method

Specification Result (µg/g)
Untreated Formaldehyde Spectrophotometeric
Limit of 3.81±2.13
Nash Reagent
detection
(LOD): 0.17
µg/g
Treated Formaldehyde Spectrophotometeric Limit of 518.37±28.84
Nash Reagent detection
(LOD): 0.03
µg/g
Treated with Formaldehyde Spectrophotometeric Limit of 495.07±25.37
Water Nash Reagent detection
(LOD): 0.11
µg/g
water and Nash Reagent detection
Vinegar (LOD): 0.17
µg/g
Water, Nash Reagent detection
Vinegar and (LOD): 0.11
Turmeric µg/g
TABLE I. Determining Levels of Formaldehyde Content of Tilapia (Oreochromis niloticus) Using Various
Solution.
Results
518.37
512.23
495.07
429.37
3.81
U n t r eat ed Tr eat ed Tr eat ed w i t h W at er Tr eat ed w i t h w at ee U n t r eat ed

an d v i n egar
Figure 1. Determining Levels of Formaldehyde Content of Tilapia (Oreochromis niloticus) Using Various
Solution.
31
Formaldehyde Absorbance Slope 0.02225

(x) (y)
0 0.06893
Intercept 0.06582
3 0.1275
6 0.20073
12 0.33293 R2 0.9999
24 0.60047
30 0.73273
Table 2. Standard Calibration Curve
Table 3. Calculated Slope, Intercept and

R2
Figure 2. A calibration curve.

Sample Aliquot Average Sample Formaldehyde Sample Average Standard
Codes HCOH absorbance mass amount Derivation
(x’) (y’)
U 4.6822 0.17 5 3.81 0.762
T 1.6099 0.03 5 518.37 115.674
TW 1.9856 0.11 5 495.07 99.014

33.308 5
TWV 4.6822 0.17 5 429.37 85.874
TWVT 1.9856 0.11 5 512.23 102.446
Table 3 Result of Concentration Content of Tilapia (Oreochromis niloticus) Using Various Solution.
32
Appendix C:
CERTIFICATION OF TESTINGS
33
34
35
36
37

Formaldehyde Degradation in Tilapia Muscles Using Various Solutions

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Amaro, M. D., Dequito, A. M., Galapon, R. G., Malubay, E. Q.

, - Formaldehyde Degradation in Tilapia (Oreochromis niloticus)

1.1 BACKGROUND OF THE STUDY

which contains essentials in fighting micronutrient deficiencies.

Fish, in densely populated countries, is considered to be significant in supplying the

the globe would require perishable food to be delivered.

1.2 REVIEW OF RELATED LITERATURE

directly to retailers than having to go to wholesalers, first.

Formaldehyde is a putrid gaseous chemical used to create building materials, as well

antiseptics, medicines, and cosmetics.

Formaldehyde occurs naturally in the environment, but may form as a result of

formaldehyde as part of normal metabolic processes (Blair, 2009).

use of formaldehyde in food is prohibited in some countries.

health conditions such as abdominal discomfort, vomiting, and renal injury.

formaldehyde in Oreochromis niloticus or Tilapia found in ponds of Freshwater Station,

Bangladesh Fisheries Research Institute (BFRI) and Mymensingh.

formaldehyde to chronic irritation and in worse cases, cancer.

that is a risk factor of excessive formaldehyde intake.

Generally, water is an effective way of removing the formaldehyde content in fishes.

of the chemical present.

In getting the accurate level of formaldehyde in Tilapia, the spectrophotometric

analysis of formaldehyde presence in abundant freshwater and marine fish species by

According to Radcliff (2018), the spectrophotometer is a device that is used to

with substances normally present as pollutants.

Nash reagent is composed of 3 components which are 2,4-pentanedione (0.2%),

Carolina states that broken up formaldehyde in aqueous samples was resolved at

submicromolar levels by derivatization with Nash's reagent trailed by fluid chromatography.

and ketones, the chromatogram is simple.

1.3 STATEMENT OF THE PROBLEM

health risks to humans.

In view of this, the researchers have decided to conduct an experimental study to

degradation of formaldehyde content in tilapia.

1.4 SIGNIFICANCE OF THE STUDY

This study determined the formaldehyde content of Tilapia (Oreochromis niloticus),

in less possible health risks caused by formaldehyde.

formaldehyde content in Tilapia (Oreochromis niloticus). The objectives of the study

1.6 SCOPE AND DELIMITATIONS

1.7 RESEARCH QUESTIONS

1. How much formaldehyde is present in tilapia based on the results of the

2. How much formaldehyde content is removed in tilapia after the following

b. Water and Vinegar

c. Water, Vinegar and Turmeric Powder

of formaldehyde content on a fish?

b. 40% Water, 60% Vinegar

c. 40% Water, 50% Vinegar, 10% Turmeric Powder

2.1 RESEARCH DESIGN

2.2 RESEARCH LOCALE

from Alabang Wet Market.

Resources - Aquatic Toxicology Laboratory including the UV-Visible Spectrophotometer

uses a quickly oscillating circular motion.

covers for easy mixing.

Diacetyldihydrolutidin complex that is measured to obtain the level of formaldehyde in the

The visible spectrophotometry method served as an indicator to detect the absorbance

principle of Beer-Lambert’s Law.

Fig.1: Vortex Mixer Fig. 2: Centrifuge

Fig. 3: Laboratory heater Fig. 4: UV – Visible

Freeze for 1 day

Use vortex mixer to mix

Fillet, mush, and weigh

Treat with 30ml