Internal Assessment:

Investigating the relationship between the cooking time and the calcium
content of kale.

IB Chemistry SL
1: Exploration

Research Question
Investigating the relationship between the cooking time and the calcium content of kale.

1.1: Introduction
Ever since I could remember, I’ve always been told to “eat my greens”. It is known that
leafy green vegetables are high in calcium and many other nutrients, which provide
many benefits to one’s body. The human body requires Calcium (Ca) to strengthen its
bones and teeth, help blood clot as well as enable muscles to contract and carry nerve
signals throughout the body. As calcium is such an essential nutrition to be absorbed by
the body, it is necessary that we consume enough of it in our daily food intake.
According to the Food and Nutrition Board (FNB) at the Institute of Medicine of the
National Academies, the Recommended Daily Allowance (RDA) of calcium for one’s
body to stay healthy is 1000mg-1300mg for teenagers and adults. This raised my
curiosity, wondering if there was a higher calcium content in uncooked vegetables as
opposed to cooked vegetables. Thus, in this paper, I decided to investigate that.

I looked at various vegetables to test and chose to conduct my experiment using kale as
it has a relatively high calcium content, with a cup of uncooked kale having an average
of 50 milligrams of calcium, according to UCSF Health medical specialists and was also
easy to obtain. The research hypothesis is that as the cooking time of the kale
increases, the calcium content of the kale as measured by the spectrophotometer and
analysed will decrease. The null hypothesis is that there will be no significant difference
in the calcium content of the kale as the cooking time of the kale increases.

1.2: Background Information

The experiment is conducted as such because of the Beer Lambert’s Law, constructed
in 1852 by August Beer, a German mathematician and chemist. According to the
Britannica Dictionary, it states that “the absorptive capacity of a dissolved substance is
directly proportional to its concentration in a solution”.
This is calculated by using the formula A = εlc.
A = absorption
ε = molar attenuation coefficient
l = path length
c = concentration
Inspired by the Beer Lambert’s law, I decided to use a spectrophotometer to find the
absorbances of the solution and from that, plot a graph of various absorbances and

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align them with their concentrations. This can be done because the law states that the
concentration and absorbances are directly proportional.
1.3: Variables
Table 1: Independent Variable
Variable Variables used Method of measurement
Cooking The sample of kale was The samples of kale were put in the oven before
time of the cooked with the cooking time, the oven was turned on. A stopwatch was used
kale measured in minutes, varying to measure the cooking time of the vegetable.
samples as follows: When the timer rang, the oven was turned off
0 (uncooked), 10, 20, 30, 40, and the kale samples were removed from the
50 oven.

Table 2: Dependant Variable

Variable Method of measurement
Calcium content The absorption of a sample of kale was tested using
of the kale spectrophotography, compared to using a calibration curve to obtain it’s
concentration, and the mass of calcium per sample was calculated.

Table 3: Controlled Variable

Variable How it was controlled Why it should be controlled
Vegetable
Type of Different vegetables will have different rates of
vegetable This was kept constant by losses of nutrients. Therefore, if various
purchasing the same type vegetables were being used, we will not be able
of kale for each condition to accurately compare the samples
from the same source
Initial mass As the calcium content will be affected by the
(±0.01g) An electronic balance was mass of the kale, it is necessary to ensure that
used to ensure that each the initial masses are constant in order to
sample had a similar initial compare the samples. It was very difficult to
mass of around 20 grams obtain a mass of exactly 20g, which is why it was
made sure that the masses were as close to 20g
as possible.
Oven
Temperature The temperature of the This is because the temperature the kale was
of oven was set to 120°C cooked at would also affect the calcium content,
the oven throughout the experiment therefore, has to remain constant when

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(±3°C) investigating the cooking time in order to
accurately analyse and manipulate the
dependant variable
Experimental Procedure
The
wavelength on The lambda max was At different wavelengths, the absorption will vary,
the found and the which is why it was crucial to keep the
spectrophoto spectrophotometer was wavelength constant throughout the experiment
meter set to that (400nm) for
every sample being tested
Solvent
Concentration The same concentration
of calcium of 0.0017g/cm3 was used Keeping the concentration of the solvent
hydroxide for all samples tested constant is necessary to ensure that the samples
can be accurately compared to each other
Volume of The same volume of 40ml Keeping the volume of the solvent constant is
calcium was used for all samples necessary to ensure that the samples can be
hydroxide tested accurately compared, as a different volume will
(Ca(OH)₂) affect the absorbances recorded on the
(±0.5ml) spectrophotometer

1.4: Apparatus and Method

Apparatus:
Kale (20g/sample)
0.0017g/cm3 of calcium hydroxide solution (40ml/sample)
Spectrophotometer (Jenway 7200)
Bunsen burner (1)
Wire gauze (1)
Tripod stand (1)
125ml beaker (6)
Glass stirring rod (1)
Conical flask (1)
Filter paper
Filter funnel (1)
Spatula (1)
55ml test tubes (6)

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Method:
Preparing the vegetables
1. Wash the kale and set aside to dry
2. Using an electronic balance, weigh each sample
and record down their initial masses
3. Cook the samples of kale in an oven at cooking
times of 10, 20, 30, 40 and 50 minutes

Preparing the sample

1. Set up a bunsen burner, tripod stand, wire gauze
and a beaker, as seen in Figure 1
2. Turn on the bunsen burner
3. In the beaker, burn the sample of kale until black
and in powder form
4. Add the sample into a 55ml test tube
5. Measure 40ml of calcium hydroxide and pour it into
the same test tube
6. Set the test tube aside on a test tube rack while
waiting for the solution to subside
7. Use filter paper, a filter funnel and a conical
1. Using the colour wheel, find the colour absorbed
and the wavelength
2. Test that wavelength range to obtain the lambda max of the wavelength, which
would be used to conduct all the tests on the spectrophotometer

Conducting spectrophotography
1. Wash the cuvette with distilled water and wipe dry
2. Add 0.0017g/cm3 of calcium hydroxide into the cuvette
3. Wipe down the cuvette with ethanol to ensure there are no fingerprints or dirt on
it
4. Insert the cuvette into the spectrophotometer to be used as the blank calibration
and set the spectrophotometer to 0
5. Wash the cuvette with distilled water and wipe dry
6. Add the filtrate into the cuvette
7. Wipe down the cuvette with ethanol to ensure there are no fingerprints or dirt
8. Insert the cuvette containing the filtrate of the kale sample for condition 1 into the
spectrophotometer
9. Record down the value displayed on the spectrophotometer

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 10. Repeat the steps for preparing the sample and conducting the
spectrophotography 4 more times with various conditions (2, 3, 4, 5 and 6)

1.5: Precautions
Safety Considerations:
- A lab coat and latex gloves were worn throughout the experiment to ensure that
the skin wasn't exposed to any harmful and damaging chemicals
- Heat source was turned off when not in use to avoid a fire breaking out
- Tongs were used when handling glassware that had been exposed to heat
- Glassware was checked for cracks and damage before usage
- Fragile glassware was handled with care to ensure it did not break
- Proper lab attire was worn, loose clothing was avoided, hair was tied back and
shoes worn were closed when working with chemicals and heat

Ethical Considerations: There were no ethical considerations taken into account.

Environmental Considerations:
- The bunsen burner and spectrophotometer was turned off when not in use to
conserve electricity

2: Analysis

2.1: Raw Data

Table 4: Unprocessed data from the experiment

Absorption/Abs (±0.001)
Average Wavelength Average
Trial Trial Trial Trial 4 Trial Absorption/
Mass per of Cooking
1 2 3 5 Abs
Cond sample/g Spectrophoto Time/min (±0.001)
ition (±0.01g) meter/nm (±0.31s)
1 20.06 400 0 2.500 2.500 2.469 2.491 2.500 2.4920
2 20.15 400 10 1.303 1.017 1.391 1.019 1.243 1.1946
3 20.03 400 20 0.791 0.842 0.783 0.800 0.772 0.7976
4 20.13 400 30 0.384 0.410 0.432 0.392 0.515 0.4266
5 20.07 400 40 0.241 0.292 0.250 0.281 0.270 0.2668
6 20.40 400 50 0.153 0.131 0.110 0.143 0.155 0.1384

Calculations for average absorption value:

- The values of the absorption of each sample of a condition is added together,
then divided by the number of samples.

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 - Example for Condition 1: (2.5 + 2.5 + 2.469 + 2.491 + 2.5) ÷ 5 = 2.492

Calculations for average mass per sample:

- The value of the initial mass for each sample in every condition (recorded in table
5), was added together and divided by the number of samples to obtain the
average mass
- Example for Condition 1: (20.09 + 20.10 + 20.03 + 19.97 + 20.11) ÷ 5 = 20.06

Table 5: Initial mass of each sample

Conditio Initial mass per sample/gram (±0.01) Average mass per

n sample/gram
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5 (±0.01)

1 20.09 20.10 20.03 19.97 20.11 20.06

2 20.11 19.94 19.92 20.03 20.05 20.01
3 19.92 20.07 20.03 19.98 20.00 20.00
4 20.00 20.03 19.95 20.09 19.93 20.00
5 20.12 20.16 20.04 20.08 20.00 20.08
6 20.00 20.11 19.90 20.03 19.92 19.99

Comment on wavelength of spectrophotometer: Fig. 3: Colour Wheel

The wavelength of the spectrophotometer was set to
400nm throughout the experiment. This was found
through observing the solution of a sample of kale
with a cooking time of 15 minutes. The colour
observed was yellowish orange, which was then
compared to the colour wheel attached in Figure 3,
showing that violet or blue was being absorbed by the
spectrophotometer. Therefore, 400nm - 500nm was
used as the range testing, in order to get the lambda
max, the maximum possible value of the wavelength.
The results were recorded in Table 6 below. When the
wavelength is 400nm, the absorption of the test
sample was the highest, therefore, 400nm was used
as the standard wavelength setting on the spectrophotometer for every test.

Table 6: Results of test for maximum Fig. 4: Graph of relationship between the
wavelength wavelength and the absorbance

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Figure 5: Graph of raw data of the relationship between the cooking time and the
average absorption, using data from Table 4.

Fig. 5 above is a visual representation of the unprocessed data from the experiment in
the form of a graph. It shows how the cooking time affects the average absorption of the
solution. The error bar does not overlap with each other, which means there is a higher
certainty in these data values. For the values with a higher cooking time, the error bars
are smaller, signifying that it is more precise than the values when the cooking time is

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less. The graph demonstrates how as the cooking time increases, the average
absorption subsequently decreases.

2.2: Qualitative Data

1) The mass of the samples decreased after cooking in the oven
2) The solutions with the samples of higher cooking times were lighter in colour and
more transparent compared to the samples of lower cooking time, as recorded in
table 7.

Table 7: Colour description of the samples of different conditions after filtration

Condition Colour description

1 Dark brownish orange

2 Dark brownish orange

3 Medium brownish orange

4 Medium yellowish orange

5 Light yellowish orange

6 Transparent yellowish orange

3) All the solutions remained the same colour scheme (yellowish orange)
throughout the experiment although some were more transparent than others
4) It was harder to separate the filtrate from the suspension using the samples with
lower cooking times. Double filtration was needed, using filter paper.

2.3: Uncertainty
Mass
The absolute uncertainty for the mass is ±0.01g
This is because 0.01 is the smallest division of the value of the mass, as the
electronic balance was able to measure the mass accurate to 2 decimal places.

Average mass
The absolute uncertainty for the average mass of each sample is ±0.01g
This was obtained through adding the uncertainty of mass for each trial together
and dividing it by the number of samples.
(0.01 + 0.01 + 0.01 + 0.01 + 0.01) ÷ 5 = 0.01

Time

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 The absolute uncertainty for the cooking time is ±0.31 seconds
This is because the smallest division that could be read by the stopwatch is 0.01
seconds, taking into account the human error and the start/stop time, which is
around 0.3 seconds. Both these values are added together to obtain the absolute
uncertainty of cooking time.
0.01 + 0.3 = 0.31

Absorption
The absolute uncertainty of absorption is ±0.001Abs.
The spectrophotometer used measures up to 3 decimal places, therefore the
smallest division is 0.001.

Average Absorption
The absolute uncertainty for the average absorption of each sample is
±0.001Abs
This was obtained through adding the uncertainty of absorption for each trial
together and dividing it by the number of samples.
(0.001 + 0.001 + 0.001 + 0.001 + 0.001) ÷ 5 = 0.001

Temperature of oven
The absolute uncertainty of the temperature of the oven is ±3°C
This is measured by dividing the smallest width of the graduation on the oven
(which is five) by two
5 ÷ 2 = 2.5
This number is then rounded to one significant figure, giving the value of 3.

Volume of Ca(OH)₂
The absolute uncertainty of the measuring cylinder used is ±0.5ml
The measuring cylinder used to measure the calcium hydroxide had graduations
of 1ml. The smallest width of graduation on the oven is then divided by two.
1 ÷ 2 = 0.5

Table 8: Percentage Uncertainty Per Condition

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 Percentage Uncertainty of Each Factor/% Total Percentage
Electronic Spectropho Measuring Uncertainty
Condition Balance Stopwatch tometer Oven Cylinder Per Condition/%
1 0.0498 0 0.040 0.25 1.25 2
2 0.0496 0.01033 0.084 0.25 1.25 2
3 0.0499 0.00517 0.125 0.25 1.25 2
4 0.0497 0.00344 0.234 0.25 1.25 2
5 0.0498 0.00258 0.375 0.25 1.25 2
6 0.0490 0.00207 0.723 0.25 1.25 2

Calculations for the percentage uncertainty of each factor:

- The values of the uncertainty for each factor is divided by the value of data for
each factor, and then multiplied by 100
- Example for the percentage uncertainty of mass for condition 1:
(0.01 ÷ 20.06) x 100 = 0.0498

Calculations for the total percentage uncertainty:

- The percentage uncertainty of each factor is added together and the answer is
then rounded off to the nearest 1 significant figure
- Example for the total percentage uncertainty for condition 1:
0.0498 + 0 + 0.040 + 0.25 + 0.12 = 1.58980, which is rounded off to 2%

2.4: Processed Data

Figure 5: Calibration curve for standard Ca2+ solution

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A calibration curve for standard Ca2+ solution in bell peppers was used to obtain the
concentration of calcium in our solutions based on the known absorbance values. The
graph shown in figure 5 is a graph obtained by SK Manirul Haque and Ahmed Abu
Judeh in the African Journal of Agriculture and Food Science Volume 1, Issue 1, 2018
(pp. 36-43) in obtaining the calcium content in bell peppers. This can be used as an
indicator of the calcium concentration in the samples of kale and is explained by the
author as the following:
“The curve was constructed by measuring the concentration and emission of several
prepared solutions, called calibration standards. Once the curve has been plotted, the
concentration of the unknown solution can be determined by placing it on the curve
based on its emission or other observable variable. The calibration curve was
constructed for calcium using the emission intensity of standard solutions. The linear
equation was y = 0.0585x with correlation coefficient r2 = 0.9935 for Ca2+”

Based on the equation of a linear curve, y=mx+c, m is the representation of the slope or
gradient. In this case, the equation is y=0.0585x, hence the m value is 0.0585.

Using the equation of the beer-lambert law, A = εlc. We can now substitute in the values
and calculate the concentration of the calcium in the samples.
A - average absorption (as found in Table 4)
ε - m value of the calibration curve (0.0585)
l - the path length (1)
c - concentration (to be found and recorded in Table 9)

Then, using the equation c = n ÷ v and n = m ÷ M, we can now substitute in the values
and calculate the mass of the calcium in the samples.
m = mass (as found in Table 5)
M = molar mass (40.08 u)
v = volume (0.04 dm3)
c = concentration (to be found and recorded in Table 9)
n = moles of substance being dissolved

Therefore, the concentration and mass of calcium was calculated and recorded in Table
9 below.

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Table 9: Calculated masses of calcium in each sample
Condition Average Concentration Volume/dm3 Molar mass/u Mass of
Absorption/ of Ca2+/ calcium/mg
Abs (±0.001) mmol/dm3

1 2.4920 42.60 0.04 40.08 68.30

2 1.1946 20.42 0.04 40.08 32.74

3 0.7976 13.63 0.04 40.08 21.85

4 0.4266 7.29 0.04 40.08 11.69

5 0.2668 4.56 0.04 40.08 7.31

6 0.1384 2.27 0.04 40.08 3.64

Calculations for converting volume from ml to dm3

- 1 ml = 1 cm3
- 1 cm3 = 0.001 dm3
- 40ml = 40cm3
- 40cm3 = 0.04dm3

Calculations for the concentration in each sample:

- Using the formula A = εlc, the concentration is found.
- Example for Condition 1:
A = εlc
c = A ÷ (εl)
c = 2.4920 ÷ (0.0585 x 1)
c = 42.60 mmol/dm3

Calculations for the mass of calcium in each sample:

- Using the formula c = n/v and n = m/M, the mass of calcium is found.
- Example for Condition 1:
c=n÷v
n=m÷M
c = m ÷ Mv
m = cMv
m = 42.60 x 0.04 x 40.08
m = 68.30 mg

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3: Evaluation

3.1: Conclusion
The results of the experiment were in favour of the research hypothesis, as the calcium
content in kale decreases as the cooking time increases. Thus, this proves a person will
obtain a higher amount of calcium from raw kale, as opposed to cooked kale, which will
benefit the health of a person. Nevertheless, kale is an excellent source of calcium even
when cooked as it still has a relatively high content compared to a lot of other
vegetables.

3.2: Strengths, Limitations and Improvement

Strengths:
- Each condition was repeated 4 more times to ensure reliability in the results with
less frequency, attempting to eliminate human errors to ensure the results of the
investigation were accurate
- The results for the total percentage uncertainty, 2%, were satisfactory, as the
uncertainty is low, making the experiment fairly reliable by displaying that the
measurements of the factors were valid
- There are many controlled variables such as the type of vegetables, initial mass
of sample, the wavelength set on the spectrophotometer, the temperature of the
oven and the concentration of calcium hydroxide to ensure that the experiment is
a fair study, and the conditions could be properly compared

Table 9: Limitations and Improvements

Limitations Improvements

A thermometer was not used, therefore, An oven thermometer should be used to

depending on the oven used, it could’ve
gotten hotter over time, taking into
account the time the oven takes to reach
120°C
obtain the exact temperature, ensuring
that the temperature of the oven
remained the same temperature
throughout the experiment and the
sample was only put into the oven once
the oven had reached 120°C

Only 5 trials were done for each condition
due to time constraints
More trials could have been done if I had
access to the lab for a longer period of
time to get more accurate average values
of the data found for the absorbances of
the kale samples.

The calibration curve used was one for I was not able to make my own calibration
calcium in bell peppers, instead of kale. curve in the lab for calcium in kale, which
is why this had to be used instead. If I
was able to, it would make the processed

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 data and the results more accurate.

The actual initial mass of the kale before
being cooked may not have been as
stated as there were natural impurities on
it affecting the mass such as sand and
water droplets, thus, making the
experiment inaccurate as the mass
would’ve been different to what was
recorded
The kale samples could have been
washed and dried several more times to
eliminate the possibility of there being
natural impurities, making the mass of
each sample more accurate and closer to
the desired mass of 20 grams.

3.3: Further investigation

A possible extension of this experiment would be investigating how the calcium content
of other vegetables vary with the cooking time. There is a possibility that the research
hypothesis set for this experiment will not work in favour with other vegetables. This
may be due to many factors such as the high presence of oxalic acid (C₂H₂O₄) in
vegetables such as spinach. Additionally, as further investigation, it would be interesting
and beneficial to also investigate how the temperature that the vegetables were cooked
at affects their calcium content. Another aspect of approaching an extension of the
experiment would be digging deeper into the effects of oxalic acid, which is present in
abundance in many green leafy vegetables such as spinach, in regards to calcium
absorption

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