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Research Question: How does the Temperature at Which Kailan Is Cooked at (25.0oC, 40.0oC, 55.0oC,
70.0oC, 85.0oC) Affect Its Calcium Content (in mg), Determined by Complexometric Titration, Using
Eriochrome Black T Indicator?

Introduction
Kailan, also known as Chinese Broccoli, is a type of vegetable. Known for its high calcium content, my mother
often cooks it for us. Sometimes she stir-fries it, sometimes she steams it, but mostly, she boils it.

One day, she came across an article detailing how different cooking methods can affect the mineral and vitamin
content of vegetables. One of the reasons for this was because of the different temperatures used in each
cooking method. I wanted to verify this fact, and while doing research online, I came across experimental
methods that could possibly be used. This sparked my curiosity; therefore, when I had the chance to investigate
this in my Chemistry Internal Assessment, I decided on the research question:
o o o o o
How does the Temperature at Which Kailan Is Cooked at (25.0 C, 40.0 C, 55.0 C, 70.0 C, 85.0 C) Affect Its
Calcium Content (in mg), Determined by Complexometric Titration, Using Eriochrome Black T Indicator?

Background Information
Calcium is a vital component of the human diet. The recommended daily intake for calcium ranges from
1,000mg to 1,200mg (Ehrlich, A.D.A.M. Editorial Team, & VeriMed Healthcare Network, 2014). Calcium is
important because it is needed for the development of bones and teeth.

Raw kailan contains 1.05mg of calcium per gram (NutritionValue.Org, n.d.), though this is often reduced when
cooking (Spritzler, n.d.). Minerals such as calcium are often leached from vegetables when they are cooked in
water (Diaz, n.d.). Even more calcium is lost when they are cooked at high temperatures – this is because
calcium binds with oxalic acid in vegetables, which are only released upon heating (Beck, 2015).

In order to find out the calcium content of kailan, complexometric titration will be used. Complexometric titration
“is a form of volumetric analysis in which the formation of a colored complex is used to indicate the end point of a
titration” (Kiruthiga, n.d.). It uses a molecule known as EDTA, Ethylenediaminetetraacetic acid, shown in Figure
1:

Figure 1. Structure of EDTA

molecule (Barak, 1998)

4-
EDTA is a polydentate ligand; it has lone pairs on all its oxygen and nitrogen atoms, and thus coordinate bonds
can be formed (Brown, Ford, & Jackson, 2014). As a result, it forms complex ions with other metal ions, such as
magnesium and calcium (University of Canterbury, n.d.):

Ca2+(aq) + EDTA4-(aq) à [Ca-EDTA]2-(aq)

Mg2+(aq) + EDTA4-(aq) à [Mg-EDTA]2-(aq)
4-
In complexometric titration, an excess of EDTA is first added into the sample solution to bond with all the metal
ions (University of Canterbury, n.d.). Afterwards, Eriochrome Black T (EBT) indicator is added, turning the
solution dark blue (its original colour).

Back titration is then carried out, using magnesium chloride solution (Middle Tennessee State University, n.d.):

H2In(aq) + Mg2+(aq) ⇋ MgIn(aq) + 2H+(aq)

(blue) (pink)

(Simplified equation: EBT(aq) + Mg2+(aq) à EBT-Mg(aq))

Initially, some of the EBT indicator (H2In in the equation above) will react with the magnesium ions from the
solution to form MgIn(aq), causing the solution to turn pink as the equilibrium position moves to the right. However,
4-
this is only temporary, because EDTA forms more stable complex ions with magnesium than EBT does
4- 2+
(University of Canterbury, n.d.). Therefore, all the EDTA molecules will displace Mg from MgIn(aq); in the end,

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the reverse reaction occurs at a faster rate than the forwards reaction, hence the solution eventually turns back
to a blue colour.
4-
It is only until all the EDTA molecules have reacted that the EBT molecules will finally form permanent complex
ions with the magnesium ions, causing the solution to turn to a permanent pink colour (University of Canterbury,
n.d.). This would then signify the end-point of the titration.

Hypothesis
Since cooking at higher temperatures will release more calcium ions from the oxalic acids, it increases the
chances of calcium being leached. High temperatures can also affect the bonding in the cell walls of kailan
(Schuhmacher, n.d.), breaking them down, hence allowing calcium ions to be more easily leached from it.

Therefore, I predict that the higher the cooking temperature, the smaller the amount of calcium content is left in
the kailan. I also predict that after a certain temperature, the calcium content of the kailan will stop declining –
this is reflected in Figure 2 when the line starts to curve and flatten. This is because I believe that there will
always be some calcium ions that are bonded very strongly to other substances in the kailan which are not
affected by heat (i.e. not oxalic acid); hence, these calcium ions will still remain in the kailan, even after cooking.

Figure 2. Sketch of my
predicted trend. (Paintbrush)

Variables
Table 1: Independent Variable
Independent variable Values I will be using How I will vary this
(±0.5oC) (oC)
25.0 (room temp.), 40.0, Water baths will be set to each of the
Temperature 55.0, 70.0, 85.0 temperatures. A thermometer will be used to
confirm the temperature inside the water bath.

Table 2: Dependent Variable

Dependent variable How I will measure this

Complexometric titration. Kailan is first cooked in water. A small sample is

taken out; EDTA, ammonia buffer and EBT indicator are added to it. The
Calcium content in kailan solution is then titrated against magnesium chloride solution until it turns from
(mg) a dark blue to a permanent pink colour. Calculations are then carried out to
find out the calcium content in the water. This is then subtracted from the
original calcium content in the kailan (before cooking) to find the calcium
content that remains in the kailan after cooking.

Table 3: Controlled Variables (for Sample Preparation)

Controlled variables How I will control this
(For sample preparation)
Using a ruler and a pair of
Surface area of kailan scissors, I will measure and
cut the kailan leaves into 1.00-
cm squares.
Significance
The higher the surface area of the kailan is,
the higher the rate it loses calcium ions to the
water. This affects the accuracy of the
results.

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Table 3 (continued): Controlled Variables (for Sample Preparation)
Controlled variables How I will control this Significance
(For sample preparation)
Use a stopwatch. I will first put test tubes of The longer the kailan is cooked, the more
water into the water bath. When they reach calcium content is lost because there is more
Duration kailan is cooked the wanted temperatures, I will add the time for water to leach the mineral from the
kailan into the test tubes and set the leaf. This affects the accuracy of the results.
stopwatch to 5 minutes. After that, the
sample will be removed and titrated.
Use a top-pan scale to measure 6.00g of If more kailan is used, then the calcium
Mass of kailan kailan. content calculated is likely to be more than it
should. This affects the accuracy of results.
3
Use a 20cm volumetric pipette to measure Using more water dilutes the concentration
3
Volume of water kailan is out 20.0cm of distilled water then add it to a of calcium content in the water. If I take out
3
cooked in test tube. only 10.0cm of kailan water for titration, the
amount of calcium content in it would be
lower. This affects the accuracy of the result.

Table 4: Controlled Variables (for Titration)

Controlled variables How I will control this Significance
(For titration)
-3
Prepare 0.05moldm EDTA solution Using a more diluted EDTA solution could mean
2+
and keep using the EDTA from the that there might be some Mg ions that are not
Concentration of EDTA same volumetric flask. To check bonded (i.e. EDTA would not be in excess). This
-3
solution (0.05moldm ) concentration, titrate against dissolved means that when EBT is added initially, it will bond
calcium tablets. with the ions and thus the initial colour of the
solution will be pink instead of dark blue. This then
makes the titration impossible.
Using less EDTA than required could also mean
2+
that there will be remaining Mg ions in the sample
3
Volume of EDTA Use a 20cm volumetric pipette to solution that are not bonded. This means that when
3 3
solution (20.0cm ) measure out 20.0cm of EDTA EBT is added initially, it will bond with the ions and
solution. thus the initial colour of the solution will be pink
instead of dark blue. This then makes the titration
impossible.
-3
Concentration of Prepare 0.025moldm magnesium If concentration used is higher, less magnesium
magnesium chloride chloride solution and keep using the chloride is added before the EBT indicator changes
solution magnesium chloride from the same to a pink colour. This affects the volume added, and
-3
(0.025moldm ) volumetric flask. thus the subsequent calculations.
EBT originally has a very dark blue when added to
2+
Volume of EBT Use a graduated pipette to measure the sample solution. When it binds with Mg , a pink
3 3
indicator (1.0cm ) out 1.0cm of EBT. coloured complex is formed. If too much EBT is
added, its darker, original colour might hide any
colour change, thus affecting the accuracy of the
titration results.
Adding more water dilutes the solution. This does
Volume of distilled Use a measuring cylinder to measure not directly affect the titration results; however, it is
3
water added to sample out 50.0cm of distilled water. less easy to tell if there is a colour change because
solution before titrating of the pale colour. Consequently, more volume of
3
(50.0cm ) magnesium chloride might be added than actually
needed, in order for a colour change to be visible.
3 1
Volume of sample Use a 10cm graduated pipette to If more sample were used, there would be more
3 3
(10.0cm ) measure out 10.0cm of sample calcium ions in the water. Hence this must be
solution to use for titration. controlled, or it would affect the accuracy of the
titration results.
3
Volume of ammonia Use a 10cm graduated pipette to Adding too much buffer solution would dilute the
2 3
buffer solution added measure out 10.0cm of buffer solution and possibly affect my ability to tell if there
3)
(10.0cm solution. is a colour change.
1 3
A 10cm volumetric pipette would have been a more precise equipment but it is not available in our school lab.
2
Ammonia buffer has a pH of about 10.5. EBT indicator will only change colours in the pH range of 7 – 11 (University of
Canterbury, n.d.), which is why it is important to add ammonia buffer to the solution.

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Apparatus
3 3 3 3
Test tubes, Clamp stand, Burette (±0.05cm ), 10cm graduated pipette (±0.1cm ), 20cm volumetric pipette
3 3 3
(±0.03cm ), 500cm volumetric flasks with stoppers (±0.25cm ), Water bath, Kailan, Distilled water, Timer
3 3 3
( ± 0.01s), 250cm conical flasks, 50cm measuring cylinders ( ± 0.5cm ), Pasteur pipette, Tweezers,
o
Thermometer (±0.5 C), Spatula, Top pan scale (±0.01g), Funnel, Weighing boat, Ruler (±0.05cm), Scissors,
EBT indicator, Ammonia buffer, Magnesium chloride hexahydrate crystals, EDTA powder, Sieve, Glass rod,
Heat resistant gloves

Calculations for Standard Solutions

Before preparing any standard solution, calculations had to be done to find out how much of the solid substance
needs to be used.

Preparing a Standard Solution of 0.05moldm-3 EDTA

-1
o Mr of EDTA (C10H16N2O8) = 10(12.01) + 16(1.01) + 2(14.01) + 8(16.00) = 292.28gmol
-3 -1 -3 -3
o 0.05moldm × 292.28gmol = 14.614gdm . For a 0.05moldm solution, I need 14.614g of EDTA
3
powder in 1.000dm of distilled water.
3 3
o 14.614g ÷ 2 = 7.307g. The volumetric flask is 0.500dm , so only 7.307g of EDTA is needed in 0.500dm
of distilled water.
o 7.307g ≈ 7.31g. Rounded off to 2 decimal points as uncertainty of top pan scale is ±0.01g.

Preparing a Standard Solution of 0.025moldm-3 Magnesium Chloride

-1
o Mr of MgCl2 ∙  6H2O = 24.31 + 2(35.45) + 6[2(1.01)+16.00] = 203.33gmol
-3 -1 -3 -3
o 0.025moldm × 203.33gmol = 5.0832…gdm . For a 0.025moldm solution, I need 5.0832…g of
3
magnesium chloride hexahydrate crystal in 1.000dm of distilled water.
3
o 5.0832…g ÷ 2 = 2.541…g. The volumetric flask is 0.500dm , so only 2.541…g of crystals are needed in
3
0.500dm of distilled water.
o 2.541…g ≈ 2.54g. Rounded off to 2 decimal points as uncertainty of top pan scale is ±0.01g.

Method3
Preparing a Standard Solution of 0.05moldm-3 EDTA
1) Using the weighing boat and top pan scale, measure out 7.31g of EDTA powder.
3
2) Add the powder into a 500cm volumetric flask using the spatula.
3) Wash the weighing boat with distilled water and pour this into the flask too.
4) Add some distilled water to the flask.
5) Swirl the flask vigorously until all the powder is completely dissolved, adding more distilled water if necessary.
Invert the flask few times to mix well.
3
6) Add more distilled water to the volumetric flask until it reaches the 500.00cm graduation mark. Ensure that
this is the actual volume of the solution and not just the reading from the meniscus.

Preparing a Standard Solution of 0.025moldm-3 Magnesium Chloride

7) Using a new weighing boat and volumetric flask, repeat steps 1 to 6 for 2.54g of MgCl2 ∙ H2O crystals.

Preparing the Kailan Sample Solution

o o 4
8) Set the water bath to 43.0 C (3.0 C higher than wanted temperature) .
3 3
9) Using the 20cm volumetric pipette, measure 20.00cm of distilled water and pour it into the test tube.
10) Put the test tube into the water bath.
11) Use the ruler and scissors to cut the kailan into 1.00-cm squares.
12) Measure out 6.00g of cut kailan using the top pan scale.
o
13) Using a thermometer, check the temperature of the water in the test tube. When it is at 40.0 C, add the
kailan into the test tube, probing them with the glass rod until they are fully submerged in the water.
14) Start the timer.
3
15) At 5 minutes, remove the test tube from the water bath and measure out 10.0cm of kailan water using
3
10cm graduated pipette. Let it cool.
16) Repeat steps 9 to 15 for four other test tubes.

Complexometric Titration
This method is adapted from the College of Science from University of Canterbury, in an experimental procedure
entitled Determination of Total Calcium and Magnesium Ion Concentration (n.d.). There are different methods for
different kinds of sample used, but I have decided to adapt the “Titration Method for Seawater, Milk and Solid
Samples” (University of Canterbury, n.d.), as opposed to “Titration Method for Fresh or Tap Water Samples”
(University of Canterbury, n.d.), because seawater and milk tend to contain more ions and are more similar to the

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kailan solution I will be using as my sample. The main modification to this method is that I am preparing my own
kailan sample solution instead of using seawater or milk.
3
17) Add the sample from step 15 to a 250cm conical flask.
3 3 -3
18) Using the 20cm volumetric pipette, measure and add 20.00cm of 0.05moldm EDTA solution.
3 3
19) Inside a fume cupboard, add 10.0cm of ammonia buffer (using the 10cm graduated pipette).
3
20) Add 50.0cm of distilled water and swirl the conical flask to mix the solution. After this, remove the flask from
the fume cupboard.
3 3
21) Using the 10cm graduated pipette, add 1.0cm of EBT indicator to the solution. The solution should now
have a dark blue colour.
22) Prepare the burette and clamp stand.
3 -3
23) Fill the burette to the 0.00cm mark with 0.025moldm magnesium chloride solution.
24) Titrate until the solution turns into a permanent pink colour. Record the end-point of the titration.
o o o o
25) Repeat the whole experiment again for the other temperatures (25.0 C, 55.0 C, 70.0 C, 85.0 C).
3 3 3
Note that the graduated pipette used to measure 1.0cm –10.0cm of solutions has an uncertainty of 1 decimal place,
3
whereas the volumetric pipette used for measuring 20.00cm of solution has an uncertainty of 2 decimal places; therefore the
number of decimal points in each volume measurement stated in my method may differ.
4
The temperature indicated on the water bath is often higher than the temperature of the solutions inside. This could be due
o
to the test tubes and large amount of water in the water bath. As a result, to cook my kailan at 40 C, I have set the water bath
to a temperature slightly higher than that.

Precautions
Table 5: Safety Precautions
Safety precautions Importance

Ammonia buffer to be used in the fume
cupboard
Keep ammonia solutions away from
open flames and heat sources.
Ammonia gas might be released during the process of adding the
buffer to the solution (University of Canterbury, n.d.). The gas is
corrosive and can irritate the lungs and eyes (CLEAPPS, 2017).
The ammonium hydroxide component of ammonia solutions will
produce ammonia gas when heated. Ammonia gas is hazardous to
health, as explained above.

Lab coats, safety goggles, and rubber
gloves are to be worn at all times.
For ammonia buffer solution, clean up
small spills with plenty of water; if it is a
large spill, ensure that the room is well-
ventilated or evacuate the laboratory
(CLEAPPS, 2017).
Be extremely careful while handling
glassware as they are fragile.
Wear heat resistant gloves when lifting
test tubes out of the water bath.
The buffer and indicator solutions are highly corrosive (University of
Canterbury, n.d.) so it is necessary to take all means to avoid
contact with skin. Eye protection is still to be worn even when
disposing (ammonia) solutions to reduce risk (CLEAPPS, 2017).
Ammonia solutions are very corrosive and are dangerous to touch.
Clearing spills with lots of water dilutes it and thus makes it less
dangerous to clean up.
Handle them carefully so they do not break, especially test tubes
and thermometers. Ensure that they are left in a test tube rack if not
in use, so that they will not roll off the table and break.
Test tubes from the water bath are likely to be hot and can burn the
skin if touched with bare hands.

Table 6: Ethical and Environmental Precautions

Ethical/ Environmental Precaution Importance

Kailan to be “treated as solid waste, although ideally it

should be composted” (CLEAPPS, 2017) – throw it into
a rubbish bin.
Solutions containing ammonia buffer solution should
be dumped into the sink in the fume cupboard. Let the
water run from the tap to rinse the sink.
Kailan might clog up the sink if dumped into it.
Ammonia gas could evaporate when emptying
the flask, and this can be hazardous to the body.
Also, ammonia “is very toxic to aquatic
organisms” (CLEAPPS, 2017) and so cannot just
be poured down regular sinks that lead to
drainages.

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Table 6 (continued): Ethical and Environmental Precautions
Ethical/ Environmental Precaution Importance

Other solutions used (e.g. EDTA, magnesium chloride)
can be poured down the normal lab sinks as they are
classed as “LOW HAZARD” (CLEAPPS, 2017).
Hazardous chemicals need to be disposed in
proper ways to prevent environmental damage.
The chemicals involved in my experiment are not
very hazardous (except ammonia solution) and so
can be poured down the lab sinks.

Raw Data
Table 7: Volume of Magnesium Chloride Solution Needed for a Colour Change at Each Temperature
Temperature (oC) Volume of 0.025moldm-3 magnesium chloride solution added5
o
(±0.5 C) (cm3) (±0.05cm3)
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
Start End Start End Start End Start End Start End
25.0 (room temp.) 0.00 28.80 0.00 27.10 0.00 29.70 0.00 29.70 0.00 31.35
40.0 0.00 29.50 0.00 28.60 0.00 28.70 0.00 29.70 0.00 26.45
55.0 0.00 29.35 0.00 26.90 0.00 27.80 0.00 26.70 0.00 27.80
70.0 0.00 27.20 0.00 26.20 0.00 23.10 0.00 21.90 0.00 27.40
85.0 0.00 25.40 0.00 25.90 0.00 26.10 0.00 21.90 0.00 27.00
5
Note that “start” means “start-point” and “end” means “end-point” of the titration

Concordant results are highlighted in the raw data table above in corresponding colours. I will use these
concordant results to calculate the average titre values for each temperature later on (page 7).

Qualitative Observations
- When preparing the EDTA solution, I noticed that there were still some powder that were not dissolved, despite
how vigorously I shook and swirled the volumetric flask.
2
- Each shredded 1.00-cm piece of kailan had a different stem structure. Some were more leafy than others.
- Initially, when added to the solution, the EBT indicator was a dark blue for lower temperatures. However, for
higher temperatures, it started off with a darker, greener colour.
o
- Although the water bath temperature was set at 43.0 C, I used a thermometer to measure the solution in the
o
test tube, which turned out to be only 40.0 C. That was also the case for the other temperatures used; the
temperature indicated on the water bath is generally slightly higher than the temperature of the solution inside.
- After 5 minutes of cooking, sample solutions were transferred out and then cooled before titrating.

Figure 3. Flask on the left shows the

colour before ammonia buffer was
added. Flask in the middle shows colour
before titration. Flask on the right shows
colour at end-point of titration. (Photo
taken by me)

Processed Data
o 3 3 3 3
There are two different sets of concordant results for 55.0 C (27.80cm and 27.80cm ; 26.90cm and 26.70cm ).
3
I have decided to use the pair of 27.80cm results because generally, my results show that as temperature
o
increases, the volume of magnesium chloride solution added decreases; hence, the results for 55.0 C should be
o 3
larger than for 70.0 C (i.e. it should be the 27.80cm pair of results).

Calculations to find average value:

• Sum up the concordant results
• Divide the sum by the number of concordant results (i.e. two)
o
• Example for 40.0 C:
3 3 3
§ 28.60cm + 28.70cm = 57.30cm
3 3 3 3 6
§ 57.30cm ÷ 2 = 28.65cm ≈ 28.70cm (rounded to nearest 0.10cm due to uncertainty )

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Table 8: Volume of Magnesium Chloride Solution Needed for a Colour Change at Each Temperature
Temperature (oC) Average volume of 0.025moldm-3 magnesium chloride solution added6
(±0.5oC)
In cm3 (±0.10cm3) In dm3 (±0.0001dm3) 7
25.0 (room temp.) 29.70 0.0297
40.0 28.70 0.0287
55.0 27.80 0.0278
70.0 27.30 0.0273
85.0 26.00 0.0260
6 3
Uncertainty (±0.05cm ) has been multiplied by two as two readings are made in each titration – start-point and end-point.
7 3 3
Note that cm was converted to dm by dividing the value by 1000.

Calculations to Determine Calcium Content

Ca2+(aq) + EDTA4-(aq) à [Ca-EDTA]2-(aq)
Mg2+(aq) + EDTA4-(aq) à [Mg-EDTA]2-(aq)
EBT(aq) + Mg2+(aq) à EBT-Mg(aq)

Using 40.0oC results as an example (Mr values taken from the IB Chemistry Data Booklet):
1) Calculate the total number of moles of EDTA used
-3 3
Ø Moles (mol) = Concentration (moldm ) × Volume (dm )
-3 3
Ø 0.05moldm × 0.0200dm = 0.001mol

2) Calculate the number of moles of MgCl2 ∙ 6H2O solution added

-3 3 -4
Ø 0.025moldm × 0.0287dm = 0.0007175mol = (7.175 × 10 ) mol
2+
Ø MgCl2 ∙ 6H2O : Mg molar ratio is 1:1
-4 2+
Ø This means (7.175 × 10 ) mol of Mg were added during the titration
2+
3) Calculate the amount of added Mg that reacted with the excess EDTA
4- 2+
Ø EDTA : Mg molar ratio is 1:1
-4
Ø Therefore this means (7.175 × 10 ) mol of EDTA molecules were in excess

4) Calculate the number of moles of EDTA that reacted with the kailan water
Ø Subtract the number of moles of excess EDTA from the total number used
-4 -4
Ø 0.001mol – (7.175 × 10 ) mol = (2.825 × 10 ) mol of EDTA that reacted in kailan water

5) Calculate calcium ion content in solution

4- 2+ 2+
Ø Molar ratio of EDTA : Mg : Ca is 1:1:1
-4 2+
Ø Hence, this means that there are (2.825 × 10 ) mol of Ca in the solution
2+
Ø Mr of Ca = Mr of Ca (electrons are so light that their mass is negligible) = 40.08
Ø Mass = Mr × moles
-4
Ø [40.08 × (2.825 × 10 )]g = 0.0113226g (× 1000) = 11.3226mg

6) Calculate the calcium content left in the kailan

Ø 6.00g of kailan were used, meaning that it originally contained (1.05mg × 6) = 6.30mg of calcium ions
(NutritionValue.Org, n.d.).
Ø Subtract the calcium content in the solution from the calcium content of the kailan.
Ø However, if the kailan only contained a total of 6.30mg of calcium ions, then it is impossible for
11.3226mg of calcium to be leached from it, as this would then mean that kailan has a negative value of
calcium ions in it.

This made me wonder about the existence of calcium ions in the distilled water I used. Therefore, I decided to
3
carry out the complexometric titration with the absence of kailan (i.e. instead of adding 10.0cm kailan solution to
3
the conical flask, I added 10.0cm of distilled water).
3 3
I did several repeats and the concordant results were 30.20cm and 30.30cm . The average of this was
3 3 3
30.25cm ≈ 30.30cm (uncertainty ±0.10cm ). Using this, I carried out similar calculations like those above, and
determined that there was 9.7194mg of calcium in the distilled water. This meant that the calculations to
determine calcium content in kailan had to take this into account. The subsequent steps for calculations are as
o
shown on the next page (still using 40.0 C results):

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7) Calculate the amount of calcium leached from kailan
Ø 11.3226mg – 9.7194mg = 1.6032mg
3
Ø This is the amount of calcium ion that was leached from the kailan in 10.0cm of solution. However, the
3
kailan had been cooked in 20.00cm of water. This means that this calcium value calculated is only half
the actual amount.
Ø Therefore we must multiply it by two: 1.6032mg × 2 = 3.2064mg

8) Calculate the calcium content left in the cooked kailan

Ø 6.3000mg – 3.2064mg = 3.0936mg ≈ 3.09mg (3s.f.)
Ø Although the concentration values of my standard solutions only have 1 significant figure (s.f.), whereas
the measurements that I made (i.e. volumes of solutions used) with the least number of significant
figures (s.f.) was 3s.f., I have still decided to round my final answer to 3s.f. as this gives a more precise
answer than 1 s.f..

9) Calculations are repeated for the other temperatures. The calculated values are tabulated below in Table 9.
(Note that the uncertainties of these values will be calculated later on in page 9).

Table 9: Calcium Content Levels Left in the Cooked Kailan at Each Temperature
Temperature (oC) Calcium Content Left in Cooked Kailan (mg)
(±0.5oC)
25.0 5.10
40.0 3.09
55.0 1.29
70.0 0.298
85.0 -2.31
8
Note that the calculated value is actually 0.288 to 3s.f.. However, since the other values in the column only have 2 decimal
places, this value has also been rounded to 2 decimal places.

Graph

Graph  1:  RelaFonship  Between  Temperature  and  the  

Calcium  Content  in  Kailan  
The graph shows a
6   downward sloping trend
Calcium  Content  (mg)  

(inverse relationship).
4   Calcium content approaches
negative values as
2   temperature increases. The
data points are all relatively
0   close to the line of best fit;
there does not seem to be
20.0   30.0   40.0   50.0   60.0   70.0   80.0   90.0   any significant anomalies.
-­‐2  
This is reflected in the
-­‐4   R²  =  0.98522   2
relatively large R value of
Temperature  (oC)  (±0.5oC)     0.98522.

Error Calculations
o
Using the 40 C results as an example, I calculated the errors and uncertainties involved that might have
affected the calculated results:

Random Error
9 10
ü Top pan scale (EDTA powder ): 0.01g ÷ 7.31g × 100% = 0.1367…%
9 10 3 3
ü Volumetric flask (EDTA solution ): 0.25cm ÷ 500cm × 100% = 0.05%
9
ü Top pan scale (magnesium chloride hexahydrate crystals): 0.01g ÷ 2.54g × 100% = 0.3937…%
9 3 3
ü Volumetric flask (magnesium chloride solution): 0.25cm ÷ 500cm × 100% = 0.05%
ü Top pan scale (kailan): 0.01g ÷ 6.00g × 100% = 0.1666…%
3 9 3 3
ü 50cm measuring cylinder (EDTA): 0.5cm ÷ 20cm  × 100% = 2.5%
3
ü Burette (experiment without kailan; start-point and end-point measurements made, each ± 0.05cm ):
3 3
0.10cm ÷ 30.30cm × 100% = 0.3300…%

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ü Burette (experiment with kailan; start-point and end-point measurements made, each ±0.05cm ): 0.10cm ÷
3
28.70cm × 100% = 0.3484…%
3 9 3 3
ü 10cm graduated pipette (sample solution): 0.1cm ÷ 10cm × 100% = 1%
ü Percentage uncertainty: 2(0.1367…%) + 2(0.05%) + 2(0.3937…%) + 2(0.05%) + 0.1666…% + 2(2.5%) +
0.3300…% + 0.3484…% + 2(1%) = 9.106…%
ü Because the value of calcium content calculated in the solution was multiplied by two, it means that the
percentage uncertainty will be twice as much too.
ü 9.106…% × 2 = 18.212…% ≈ 20% (percentage uncertainty to be rounded to 1s.f. for values above 2.0%)

Absolute Uncertainty
ü 3.09mg (±20%)
ü 20% × 3.09mg = 0.618mg ≈ 0.62mg
ü 0.618mg is rounded to 0.62mg as 3.09mg has 2 decimal points
ü However, the final value must be a multiple of the uncertainty, hence I must round 3.09mg to 3.10mg
ü 3.10mg (±0.62mg)
ü Number of decimal points in results need to be equal to number of decimal points in uncertainty value.
9
These error values will be multiplied by two. This is because in order to calculate the calcium content of the solution, the
experiment had to be carried out twice – once with kailan water as the sample solution and once without kailan (to find
out the calcium content of distilled water). As a result, some of the random error will occur in both experiments, hence
needs to be multiplied by two.
10
Note that although EDTA solution was used in excess, the value of its concentration was still used in calculating the
amount of calcium content remaining in the kailan, thus I have also included its uncertainty in my error calculations.

Other Temperatures:
Random Error
o
ü 25 C = 18.188…% ≈ 20%
o
ü 55 C = 18.234…% ≈ 20%
o
ü 70 C = 18.247…% ≈ 20%
o
ü 85 C = 18.284…% ≈ 20%
ü Rounded to 1s.f. since values are above 2.0%

Absolute Uncertainty (Final values rounded accordingly to uncertainty values)

o
ü 25 C = 5.10mg (±1.02mg)
o
ü 55 C = 1.30mg (±0.26mg)
o
ü 70 C = 0.30mg (±0.06mg)
o
ü 85 C = -2.30mg (±0.46mg)

Conclusion
According to my data, there is an inverse relationship between the cooking temperature and calcium content left
in the kailan. This is evidenced by the graph, which shows a straight line of best fit with a negative gradient.

The higher the cooking temperature, the lower the calcium content remaining in the kailan. This is due to the fact
that more calcium ions have been leached from the vegetable by the water. At the lowest temperature
o
investigated, 25.0 C, only 1.20mg of calcium was leached into the solution, meaning that the cooked kailan had
retained about 81% (5.10mg out of 6.30mg) of its calcium content. However, at higher temperatures such as
o
70.0 C, almost all the calcium minerals seem to have been absorbed from the vegetable, suggesting that we
o
should not cook kailan in water for temperatures above 70.0 C, as it loses its nutritional value. A reason for this
inverse relationship between temperature and calcium content in kailan could be because at higher
temperatures, calcium ions are more soluble in water. Water molecules possess more kinetic energy, hence they
collide at a higher frequency with the calcium ions in the kailan, hence leaching occurs at a faster rate. As a
result, in the same time period of 5 minutes, more calcium is lost from the kailan.

My hypothesis is correct, in the sense that at higher temperatures, calcium content remaining in kailan will be
lower. However, it seems that I was wrong about how there would be a limit as to how much calcium content can
be leached from the kailan. Unlike my prediction, my results did not have a curve of best fit, but instead, a linear
one.
2
Nonetheless, the line of best fit has a relatively high R value of 0.98522, which suggests that the trend it shows
should be quite reliable. However, I am not confident in it, because the line slopes downwards into the negative
y-axis values. This is problematic because it is impossible for kailan to have a negative amount of calcium
o
content. The experimental results for 85.0 C also produces an unachievable value of calcium content (-2.30mg),
which makes me doubt the reliability of my results for higher temperatures. Then again, it could be the source
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which provided an inaccurate literature value of 1.05mg per gram of raw kalian (NutritionValue.Org, n.d.), though
given the nature of experiments and the credibility of the website, it is more likely that something had went wrong
in my experiment.

Although my results seem somewhat reliable for the lower temperatures, I have many reasons to doubt my
results for higher temperatures. Firstly, as mentioned in the Qualitative Observations section, the EBT indicator
had a darker and greener initial colour when added to solutions with higher temperatures. This made it more
difficult to notice the colour changes to pink, which would determine the end-point of the titration. It was easier to
notice colour changes for lower temperatures, because the initial colour of the EBT indicator upon addition was a
more solid blue. Secondly, although the random errors of their respective temperatures all round up to 20%, if
we look at the unrounded percentages, the random error actually increases as the temperature gets higher. This
suggests that the results get less and less precise as the temperature increase, though the random error only
increases by a small amount between each temperature.
2
Despite having concordant titration results, a large R value, and a trend that accords to my hypothesis, I am not
very confident in my overall results. The reason for this being that 20% is a relatively high random error, though
there is not much I can do about this because I have already used the most precise equipment available in the
lab for all my measurements. Additionally, I have kept all my controlled variables constant in each repeat, so if
my results for lower temperatures are plausible, then it should be for the higher temperatures too. But it was not,
and this implies that I may have high systematic errors in my experiment.

I am unable to pinpoint the exact cause of the high systematic error, but it is quite possible that magnesium ions
are more soluble than calcium ions (kailan contains both magnesium and calcium ions). As a result, at higher
4-
temperatures, significantly more magnesium ions will be leached into the solution from the kailan; the EDTA
2+ 4-
ions may then form complex ions with Mg , hence more EDTA are used up. Consequently, less magnesium
chloride solution is needed before the solution turns pink during the titration process. It may then seem as if this
4-
is because there were more calcium ions in the solution, resulting in many of the EDTA ions forming complex
coordinates, when it is actually because of the magnesium ions. Consequently, this affects the accuracy of my
results; this is a potential explanation for the high systematic error in my experiment, thus my lack of confidence
in the accuracy of my results.

Regardless, I believe that the trend is still somewhat reliable – the fact that at higher temperatures, calcium
content in the kailan decreases. This is because it makes sense scientifically, and if I did have a high systematic
error, it affects all the results across all temperatures, meaning that even if my results are all inaccurate, the
trend they show should still be relevant. Therefore, this means that in future, in order to minimise the loss of
calcium from kailan, my mother should cook it at lower temperatures.

Evaluation
Table 10: Strengths of My Experiment
Strengths
Put a white piece of paper underneath the
flasks so that colour changes were more
noticeable.
Repeated all experiments 5 times,
ensuring that I had at least a pair of
concordant results for each temperature.
Inverted the volumetric flask of EDTA and
MgCl2 solutions before using each time.
3 3
I cooked the kailan in 20.00cm of water
3 3
then measured out 10.0cm of solution for
titration.
Significance
The table was a dark blue colour, hence it was quite hard to tell if
the solution has turned pink during the titration – this could lead to
larger-than-actual values of titration results. Using a white piece of
paper makes the colour changes more noticeable.
Concordant results should be used to calculate averages for
titration to increase the reliability of the results.
Ensures that the particles are more evenly distributed throughout
the solution.
To ensure that all the kailan was cooked, all the shredded pieces
3
needed to be completely submerged in the water. 10.0cm of water
was too little for this. I decided to cook kailan in 20.00cm of water
but re-measured 10.0cm for the titration because after cooking the
kailan, it is likely that some of the water will evaporate, causing a
decrease in the total volume of solution. Therefore, measuring the
volume of solution to use just before titrating it will provide more
accurate results.

Used a thermometer to check the Ensures that the temperature of the solution is at the wanted
temperature of the solution in the water temperature, because sometimes the water bath might indicate a
bath. higher temperature than the solutions placed inside are at.

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Table 11: Weaknesses and Limitations in My Experiment
Weaknesses/ Limitations Significance Possible Improvements

Although I repeated my Concordant results are used to
experiment 5 times and calculate averages for titration. The
ensured that I had at least a more concordant results I have, it
pair of concordant results for means my results are more reliable.
each temperature, my Thus, my final calculations will also
calculations would probably be likely to be more accurate.
have been more accurate
and reliable if I kept
repeating and collected more
concordant results.
Time elapsed between If more time elapses, this means that
taking the test tube out of the there are more chances for more
water bath and transferring calcium content to be leached from
the sample solution out to let the kailan into the water. It also
it cool. means that kailan are essentially
cooked for slightly longer than
5minutes.
At lower temperatures, when The darker, greener colour made it
the EBT indicator was added hard to tell if there was a permanent
to the solution, it was a dark pink colour in the solution, meaning
blue. But for higher that results collected for higher
temperatures, EBT had a temperatures are likely less accurate
greener colour. than for lower temperatures.
3
It was difficult to measure 1.0cm of
EBT indicator has a very EBT indicator because of its dark
dark colour. colour. Nonetheless, this does not
necessarily affect the reaction too
much because EBT acts as an
indicator and is not involved in the
calculations process.
If there are significantly more [Mg-
2- 2-
It is impossible to tell what EDTA] ions than [Ca-EDTA] , less
2-
the ratio of [Mg-EDTA] to MgCl2 solution would be added
2-
[Ca-EDTA] is in the before the solution turns pink during
solution. titration. During calculations and data
interpretation, it would seem as
though more calcium ions were
4-
leached, causing more EDTA
molecules to be complexed, when it
was, in fact, not.
Keep repeating until I have 3–5
concordant results, instead of finishing
the experiment at 5 repeats, and a
pair of concordant results at each
temperature.
Get the conical flask and place the
sieve on it (to filter out the kailan
pieces). At 5minutes, immediately
remove the test tube from the water
bath and pour the contents into the
conical flask.
Use other complexometric titration
indicators such as calmagite, which
has a brighter colour, which makes
colour changes more noticeable. I
could also use colorimetry to
determine the concentration of colours
to ensure that the intensity and
wavelengths are the same (i.e. same
shade of pink) at the end-point of each
titration.
When using EBT indicator, use a
burette to measure the volume. Fill the
burette till the meniscus is at the
3 3
0.00cm mark, then add 1.00cm of
indicator. Always read from the
meniscus when using dark-coloured
chemicals.
Take the solubility product constant
(Ksp) of each type of ion into account
during calculations, or use atomic
absorption spectroscopy to determine
the calcium content in the solution
(Colorado State University, 2017).

Extensions
The experiment could be repeated using other calcium-rich sources such as kale, watercress, or even milk,
to see if there is still an inverse relationship between calcium content and temperature. Additionally, the
same experiment can also be repeated using other temperatures (e.g. 10.0oC, 100.0oC), to ensure that this
relationship still corresponds. In order to further extend my understanding, I can also carry out this
experiment for other minerals in vegetables such as iron and/or magnesium.

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http://soils.wisc.edu/facstaff/barak/images/chelate.htm

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Spritzler, F. (n.d.). How cooking affects the nutrient content of foods. Retrieved May 8, 2017, from
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Acknowledgements
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