Chemistry Internal Assessment Standard Level

To what extent does the repeated heating of vegetable cooking oil (1H,5H,10H) affect the
content of total free fatty acid?
Candidate number: lcm 051
May 2024 Session

Introduction

When I went to a restaurant, my family and I found that the oil the restaurant used had been cooked
repeatedly. When we saw it, we immediately left the restaurant, however, my uncle said that it was ok
as they used vegetable oil. Hence this sparked my interest, as to how different types of oil make the
decision justifiable. After the event, I tried to search what is the determining quality assessment in oil,
which turns out to be free fatty acid. Free Fatty Acid is the fuel for most of the body, excluding the
brain, medulla, and erythrocytes. It affects the body’s hormonal secretion and has many effects on
glucose homeostasis. Fatty Acid is often divided into two, which are Polyunsaturated Fatty Acid and
Saturated Fatty Acid. Polyunsaturated Fatty Acid would be better than saturated Fatty Acid as
Polyunsaturated Fatty Acid does not deposit on the artery while Saturated Fatty Acid does, causing
the artery to be clogged. Due to that, I wonder if there will be a difference in the composition of the
fatty acids in different vegetable oils and if heating them may make any differences. Therefore, I
decided to make my focus my research on answering the question “ To what extent does the
repeated heating of vegetable cooking oil (1H, 5H, 10H) affect the content of total free fatty
acid? ”

Background and Method

1. Vegetable Oil

Vegetable oils are oils that is sourced from seeds, nuts, or grains from vegetables. Vegetable
oil is known to be high in unsaturated fatty acid in contrast with animal fat which is high in saturated
fatty acid. The main differences between those fatty acids would be that saturated fatty acids have no
double bonds with the general formula of CH3(CH2)nCOOH, while the unsaturated fatty acids have
one or more double bonds known as oil.

Figure 1: Saturated and Unsaturated Fatty Acid

However, both of them are known to contain triglycerides that is an ester with three acids unit that
builds the adipose tissue and sebum
The saturated fatty acid in contary to polyunsaturated fatty acid is not an essential fatty acid as
it can be produced in the body. Polyunsaturated fatty acid is also needed in the body as it makes the
transport of cholesterol more efficient, while the saturated fatty acid increases HDL cholestrol which
may worsened the body condition.

2. Acid-Base titration

Figure 2: Acid-Base titration procedure

Acid-base titration is needed to determine the amount acid and basic solution in the solution.
For it to be working, it needs titrand and reagent. Where titrand is the solution with unknown molarity,
while reagent is the solution with known molarity that needed to calculate the molarity of the titrand.
In order to do this, an analyte is prepared by mixing it with the solution. Or in this case the oil
with ethanol to make an acidic solution. It will then be titrated by a known molarity of NaOH that is
0.1 N. It will then titrated until the end point as it changes colour because of the indicator

3. Indicator

Figure 3 : Phenopholtalein affected solution

pH indicator is a tool to seee the endpoint of titration. It is a solution that changes color as
quick as it closes the pKA or the acid constant. The indicator is needed in small amount as higher
amount will affect the accuracy of the reaction. The indicator needed in this experiment would be
phenopholtalein as it changes color from colorless to pink near it’s pKA.
 Independent Variable Types of oil used (vegetable oil, especially soybean,
and palm oil).

Dependent Variable Amount of Free Fatty Acid in each sample.

Controlled Variable How It is Controlled Significance

Mass of the Samples The mass of each sample would be For oil’s acid value to be edible,
20 grams it should be less than one.
Hence, the weight should be 20
grams1.

Concentration of NaOH The concentration of NaOH would Removing the impurity of sulfur
be 0.1 M, with a mass of 4 grams in oil, 0.1 M is used for
and a volume of 1000ml efficiency.

Concentration of Ethanol The ethanol used would be pure Ethanol is a sufficient solvent for
ethanol with a concentration of acid-base titration. 96%
96% concentration would ensure the
purity of the sample.

The amount of The amount of phenopholtalein Acts as the pH indicator and will
Phenopholtalein solution solution used would be 2-3 drops appear pink in the basic solution.
per sample.

Frequency of heating The frequency of heating would The amount of heating is

be one, two, five, and ten times referenced to an already done
experiment by

Period of heating and The heating process will be with a The period of heating and
cooling temperature of 180 degrees and a cooling is used to standardize the
cooling down of 5 hours treatment of the oil samples

Storage The oil samples were kept at a Preventing the oil oxidation and
minimum temperature in places of light and temperature
with minimum light exposure exposure

Table 1: Variable used

Apparatus and Chemical Used:
● NaOH (0.1M)
● Ethanol (150 mL)
● Oil (20 g)
● Phenolpholtalein (2-3 droplets)
● Distilled water (1 L)

● Analytical Balance ± 0. 01 ml
● Burette (with stand) ± 0. 02 ml

1
(MicroChem's experiment, 2020)
 ● Measuring Cylinder± 0. 5 ml
● Conical flask± ± 2. 5 ml
● Hot Plate
Method:

1. Dissolve 2 grams of phenolphthalein in 100mL of ethyl alcohol and mix well by shaking.
2. Dissolve 4g sodium hydroxide pellets into 900 ml of distilled water, cool, and make a final
volume of 1000 ml.
3. Take 20 grams of oil in the conical flask
4. Measure 50 ml ethanol (96%) and pour it into another conical flask(250ml).
5. Add 2-3 drops of pp indicator to the ethanol, and titrate using 0.1 N NaOH until the solution
turns light pink
6. Now, the neutralized ethanol is added to the flask containing the sample, and shake the flask
to mix the solution.
7. Boil the mixture by placing the flask on a hot plate until the sample dissolves in ethanol
completely.
8. Continue the boiling until the sample is dissolved completely.
9. Shaking with hands should be continued periodically.
10. Note the Final Burette reading

Considerations

Chemical Substances Risk Hazard Safety Precaution

Phenolphthalein May cause skin and eye irritation, Wear appropriate goggles,
and may damage the digestive clothes, and gloves. Avoid direct
tract if swallowed contact with eyes and skin.
NaOH May irritate if in contact with the Always wear protective gloves
skin and eyes and goggles. Avoid contact
with Ethanol Corrosive to skin in contact, Use gloves when handling, and
inhaling high concentration may use a mask to avoid breathing.
affect the lung
Distilled Water No risk hazard No necessary precaution needed
Glassware May cause harm to skin and eyes Follow the procedure in the lab.
if broken. Avoid bare-hand contact with
glass.
Table 2: Safety Consideration
Ethical Considerations: The oil is ethically sourced from palm trees and soybeans, and there is no
animal or dairy product involved. However, the ethical consideration would be for the underpaid
workers and the recent rise of land opening for oil. Hence, the use of oil will only be for as much as
needed.

Chemical Substances Safety Precaution

Oil Pour oil into the non-recyclable package, and
throw away the oil
NaOH Throw away the NaOH down the drain as long as
it does not exceed the pH of 12.
Ethanol Throw in a specialized gallon for hazardous waste.
Phenolphthalein Dilute it with a lot of water and drown it down the
sink

Table3: Environmental Consideration

Results and Discussion

Fresh Oil The volume of NaOH (ml )

Type of Oil Burette Trial 1 Trial 2 Trial 3

Readings

Soybean Oil Initial 48. 88 𝑚𝑙 48. 57𝑚𝑙 48. 25 𝑚𝑙

Final 48. 57 𝑚𝑙 48. 25 𝑚𝑙 47. 95𝑚𝑙

Palm Oil Initial 50. 00 𝑚𝑙 49. 61 𝑚𝑙 49. 25 𝑚𝑙

Final 49. 61 𝑚𝑙 49. 25 𝑚𝑙 48. 88 𝑚𝑙

1H Oil The volume of NaOH (ml )

Type of Oil Burette Trial 1 Trial 2 Trial 3

Readings

Soybean Oil Initial 48. 88𝑚𝑙 48. 31𝑚𝑙 47. 69𝑚𝑙

Final 48. 31𝑚𝑙 47. 69𝑚𝑙 47. 05𝑚𝑙

Palm Oil Initial 47. 05𝑚𝑙 46. 6𝑚𝑙 46. 14𝑚𝑙

Final 46. 6𝑚𝑙 46. 14𝑚𝑙 45. 66𝑚𝑙

 5H Oil The volume of NaOH (ml )

Type of Oil Burette Trial 1 Trial 2 Trial 3

Readings

Soybean Oil Initial 45. 66𝑚𝑙 45. 02𝑚𝑙 44. 38𝑚𝑙

Final 45. 02𝑚𝑙 44. 38𝑚𝑙 43. 73𝑚𝑙

Palm Oil Initial 43. 73𝑚𝑙 43. 13𝑚𝑙 42. 52𝑚𝑙

Final 43. 13𝑚𝑙 42. 52𝑚𝑙 42. 34𝑚𝑙

10H Oil The volume of NaOH (ml )

Type of Oil Burette Trial 1 Trial 2 Trial 3

Readings

Soybean Oil Initial 42. 34𝑚𝑙 41. 83𝑚𝑙 41. 19𝑚𝑙

Final 41. 83𝑚𝑙 41. 19𝑚𝑙 40. 61𝑚𝑙

Palm Oil Initial 40. 61𝑚𝑙 39. 85𝑚𝑙 39. 09𝑚𝑙

Final 39. 85𝑚𝑙 39. 09𝑚𝑙 38. 31𝑚𝑙

Table 4: Raw Data

Processed Data

Steps Explanation

Knowing the difference between each trial The different concentration is needed as it
represents the TFA (Total Fatty Acid) content in
𝑑= 𝑉1 − 𝑉2 each sample.
48. 88 − 48. 31𝑚𝐿
0. 087𝑚𝐿

Finding the mean (𝑀) The mean value is used to find the center score
and will be used as the representation /sample.
𝑉1+𝑉2+𝑉3
𝑀= 3
0.087+0.090+0.084
= 3
= 0. 087 𝑚𝐿
Finding the standard deviation (SD) It is used to summarize continuous data and it is
used to calculate a research sample.
2
Σ𝑥
𝑆𝐷 = 𝑁
2
Σ0.087
= 3
2
Σ0.087
= 3
= 0. 003

Finding the relative standard deviation It is used to measure the dispersion of the set of
(RSD%) numbers around the mean, the relative standard
dispersion of around 5-20% is the
𝑅𝑆𝐷% =
𝑆𝐷
× 100 recommended value.
𝑚
0.003
= 0.087
× 100
= 3. 45%
Table 5: Data processing

Type of Oil TFA value Mean SD RSD%

Fresh Oil Trial 1 Trial 2 Trial 3 Mean SD RSD%

Soybean Oil 0.087 0.090 0.084 0.087 0.003 3.45%

Palm Oil 0.109 0.101 0.126 0.112 0.013 1.14%

Type of Oil TFA value

1H Oil Trial 1 Trial 2 Trial 3 Mean SD RSD%

Soybean Oil 0.160 0.174 0.179 0.171 0.001 5.76%

Palm Oil 0.126 0.129 0.134 0.129 0.004 3.12%

Type of Oil TFA value

5H Oil Trial 1 Trial 2 Trial 3 Mean SD RSD%

Soybean Oil 0.180 0.180 0.182 0.180 0.001 0.905%

Palm Oil 0.168 0.171 0.18 0.173 0.006 3.61%

 Type of Oil TFA value

10H Oil Trial 1 Trial 2 Trial 3 Mean SD RSD%

Soybean Oil 0.180 0.180 0.16 0.173 0.011 6.65%

Palm Oil 0.214 0.214 0.219 0.215 0.028 1.36%

Table 6: Processed Data

Quantitative Analysis

Upon observing the samples, it is seen that the color of the soybean oil didn’t change drastically.
However, the change in the palm oil is noticeable as it generates more hay in the oil. However, both
oils are still looking clear enough and not as black as I expected. When I titrated it however, I noticed
that some of the samples looked to be more pink than I expected and the pink persisted longer than 30
seconds, which indicates that some of the samples were overshot with KOH.

Figure 4: Titration Result

Statistical Analysis

I will now analyze the statistics of my data using a bar chart, as a bar chart is useful in showing the
changes in the data over time. The data that I got was

Graph 1: How heating affects acid value in soybean oil Graph 2: How heating affects acid value in palm oil
It can be shown that the R-value is around 0. 614 which shows a moderately strong, while the R-vale
of the Palm oil’s TFA is around 0. 969 which shows a very strong correlation. However, it is shown
that there is an outlier in the first heating where the TFA value is not in the range of the error bar. It
seems that there is an error in the storing or heating process which affects the acid value greatly.
However, the total fatty acid amount in palm oil is really good and does not differ from the error bar.
Fortunately, I am searching for a trend in the data which is that reheating oil affects the total fatty acid
which is proved. Both of the data show a positive correlation

However, it can be viewed that in the data of soybean oil, the standard deviation is spread more than
the palm oil. Hence, making the standard deviation not touching the gradient line that shows the trend.
The reason may be due to with the lack of precision because of human error. The error may come
from many sources, especially in preparing the sample of titration which have to do with calculation
error. The error may also come from the titration which may become overtitrated as some of the
reactions result in brighter colours as light pink that persisted for more than 30 seconds. It could also
be because of the mistake of the oil storing process in which I used previously used oil. Hence,
making the titration inaccurate

Literature Review

A similar research has also shown a similar acid value to heat and trend that can support my finding
(Zoltan Szabo, 2022). The possibility of the oil having been oxidized was also addressed by research
(Oils Fats NZ, 2023).

Uncertainty Calculations

Tools Used Absolute Uncertainty Percentage Uncertainty

Digital analytics was used to calculate 20 ± 0. 001 =0. 001/20×100

grams of oil =0. 005

Tools Used Absolute Uncertainty Percentage Uncertainty

Burette was used to calculate the amount ± 0. 02𝑚𝑙 = 0. 02/50 × 100

of KOH used = 0. 04

Measuring cylinder was used to calculate ± 0. 05 𝑚𝑙 = 0. 05/100 × 100

the volume of phenopholtalein solution = 0. 05
 Measuring Cylinder was used to calculate ± 0. 05 = 0. 05/1000×100
the volume of KOH solution =0. 005

Table 6 : Uncertainty Calculation

Conclusion

In the end, it is safe to say that the amount of fatty acid correlates positively both in soybean and palm
oil. However, the amount of fatty acid in soybean oil is not making a trend as drastic as palm oil
which shows a positive correlation. However, according to the experiment, the amount of acid values
in each value of palm and soybean oil are around the same values. However, the error percentage in
the scientific research was found to be smaller than the error in the oil. It is only natural because of the
set conditions that have been set for the sample oil that are stored in dark and cold environments, and
the pink color measured to be only 30 seconds appearing in the sample, which increases the accuracy
greatly.

Even though the results are around the same, the results are not yet accurate as the percentage error is
still somewhat large. The main accusation would be that it is the result of systematic error and human
error that make the result to be flawed.

However, after searching for another source, I have found that the acid oil in soybean oil is also
greatly affected. It is then found, that the result is actually because of the media of the frying which is
using real meat for the experiment which is the main thing that people fry. Hence, it can be said that
the result of the fatty acid is only applicable when frying a potato and is not recommended for any
other objects being fried

However, answering the research question it is assured that even frying the oil so that it wouldn’t burn
and frying vegetables still have a significant effect on oil quality. Hence it is not recommended for
anyone to eat foods that are the result of frying repeatedly

Weaknesses

The first weakness to be found would be that the experiment is carried out for multiple days, it is the
opens the possibility of the oil oxidized and exposed to heat or light which nay potentially affect the
quality of the oil. It can be found in one of the most recent data where the amount of RSD% is 6.31
which is considered high for an acid value experimet
The second weakness would be the disparity between the acid value percentage error and the actual
error. Hence, it is needed for the tools to be evaluated again and it would be the result of systematic
error instead of human or random error.

Systematic Error

Titration Endpoint

the titration is being done on a scale of 0.1 scales. It is then recognized that the experiment is prone to
error and sometimes overshot. Hence, I can say that some of the samples were found to have the color
of pink in more than 40 seconds and one of the samples was too purple to even be considered an
accurate titrant. However, for the titration to be accurate it is needed for some people to watch over
the titration or for one person to drop the KOH and one person to shake the samples.

Dissolving the Oil

Some errors may also occur because of the flaw in dissolving the oil. The oil sample needs to be
shaken periodically and heated with ethanol. It is then said that the solution may be prone to boiling
and the lack of shaking may also mess up the value of the acid values itself.

Random Error

Some errors that are unwatchable and out of the experiment's control may also occur, such as air
contamination, the effect of contact with the body, or even as simple as the excretion product contact
the sample itself. To reduce this random error, the experiment needed more people to watch over and
monitor, and the second is to always have protective equipment so that the sample could be kept
sterile.

Strengths

One of the strengths that I can be proud of for my result is how small the disparity and uncertainty in
the calculation. I am proud of my display of acknowledging the uncertainty in all the analytical and
glassware tools to make an accurate calculation. I am also aware of using electronic tools which also
improve the accuracy of my results. The second would be the use of aquades and how I managed to
keep all my samples sterile and clean for the usage of further experiments.
Extensions

After learning about titration, I have now developed a specific liking for the titration process,
especially on oil and such. I am also interested in exploring the other types of titration such as acid
and base titration and metal titration. As for the experiment, it is suggested that for further
experiments, the amount of oleic and palmitic acid can be broken down to see the actual result of the
heating of soybean oil. As the oil that has been heated though may not have a large difference still
produces a free radical product that also contributes to the oil quality.
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