Introduction

The human body needs various vitamins to be able to

carry out all the important biological functions in our
body. The human body cannot synthesise vitamin
molecules (which is interestingly common in primates as
well but not in other mammals, such as cats and dogs as
they can synthesise their own Vitamin C), and therefore
the vitamins must be digested through our diet. When
there isnt a particular vitamin in our diet, a particular
disease, that is associated with the lack of that vitamin
would result.
Vitamin C is known as ascorbic acid and has the structure like the picture at the
side. Vitamin C can be found in various fruits and vegetables such as; oranges,
other citrus fruits, vegetables, strawberries and green vegetables. The objective of
this experiment is to measure the amount of Vitamin C in each of the orange juices
and to see if they match the amount of Vitamin C mentioned at the side of the
packaging. Vitamin C decolourises the blue dye DCPIP (dichlorophenolindolphenol).
Vitamin C is an antioxidant and reduces the DCPIP (Vitamin C acts as a reducing
agent as it is easily oxidisable). DCPIP changes from blue to colourless (or slightly
pink) as it becomes reduced. An antioxidant is a substance such as Vitamin C that
removes potentially damaging oxidizing agents in a living organism.
Exposure to oxygen can cause some atoms to convert to a free radicals(which is
an atom that has an unpaired electron and this can cause a chain reaction. The free
radicals would react with any molecule and latch onto the electron of another cell
and this can cause a change in the formula and structure of the cell, causing there
to be less of that molecule, resulting in a change in the cell structure, which could
cause the DNA in that cell to change. This can cause serious diseases such as;
cancer, heart disease,
stroke, aging,
diabetes, arthritis,
fibromyalgia,
Parkinsons Disease,
autoimmune disease
and so on. An
antioxidant like
Vitamin C can donate
an electron to a free
radical, causing it to
not be a free radical
anymore. This then
stops the reactions
from occurring, thus
leaving the cell and its
structure and not
altering it in any way.
Antioxidants can also
repair the cell damage made by the free radicals. This means that Vitamin C
reduces the risk of cancer.

Vitamin C is also essential for the formation of collagen, which is the protein that is
found in bone and connective tissue such as tendons and cartilage. Vitamin C also
helps to maintain the integrity of skin, connective tissue, bone, blood vessels, and
your gums. A deficiency in Vitamin C can cause a person to develop a disease called
Scurvy, in which the body is unable to repair and synthesise collagen, leading to
abnormal bone and dentine formation and, occasionally, haemorrhages. The
recommended daily amount of Vitamin C for the average adult (according to the US
Government) is at 75 to 90 mg per day. It is, however, debatable if Vitamin C that is
ingested in large amounts, on a daily basis, could be harmful to life as an overdose
could have unexpected effects.

Hypothesis:
The higher the amount of vitamin C in the juice, the faster the DCPIP would be
decolourised and the orange juice should have the highest amount of Vitamin C as it
does not have any sugar or carbohydrate mixed into it.

Equipment:
Burette,
Different types of orange juice,
Orange,
DCPIP,
Different concentrations of VItamin C,
Funnel,
Clamp Stand,
Measuring cylinder,
Pipette,
Conical Flask,
Cloth to filter the orange juices,
Pestle and Mortar (to squeeze the juices from orange juice)

Ethics:
There are no ethics considering this experiment besides the fact that the orange
juice used could be used as a food source rather than being used in this experiment.

Plan:
Creating a calibration curve
1. Set up a burette in a clamp stand - you will be performing a titration.
2. Put 3 ml of DCPIP into a small conical flask and place this on a white
tile underneath the burette tap.

3. Ensuring the tap is closed in the burette, add 20-30 ml of a known

concentration of ascorbic acid (vitamin C) to the burette. Use a funnel at the
top to ensure no splashes.
4. Make a note of the start volume on the burette - preferably in a table.
5. Slowly, add the ascorbic acid until the DCPIP decolourises.
6. Make a note of the end volume on the burette. Work out the volume
added.
7. Repeat another 2 times with the same concentration, ensuring 3ml of
DCPIP is added to a rinsed conical flask each time.
8. Repeat steps 1-7 with the other 5 concentrations.
9. Draw out a calibration curve on graph paper and draw a best fit line for
the graph.
Testing the juices
1. The procedure is the same, only this time you must add filtered juice to the
burette.
2. Record the volume needed to decolourise the DCPIP - 3x for each juice so that an
average volume can be taken.
3. Using the calibration curve, work out what the % if Vitamin C is for each orange
juice.
4. To create the extract from fresh orange juice - mash the orange juice and filter
the pulp.

Change: The type of orange juice (the orange juice sources)

Organism: Orange juice
Repeat 3 times to ensure reliable results
Measure: The volume of orange juice needed to decolourise DCPIP
Same: The volume of DCPIP used, the type of fruit used(orange juice), temperature
of solution, concentration of DCPIP solution, the same end point.

Results
Concentration of Vitamin
C/Type of Orange juice

Amount of Vitamin C needed to decolourise the DCPIP (ml)

Start

End

Change

Average

21

21

21.5

44

22.5

20.1

42.2

22.1

21.7

21.7

21.7

43

21.7

28.7

48.8

20.1

13.9

13.9

13.9

27.3

13.4

27.3

41.6

14.3

12.3

12.3

12.3

24.2

11.9

24.2

35.2

11

6.5

12.5

12.5

19.8

7.3

19.8

25.8

1.9

2.4

0.5

2.4

2.9

0.5

2.9

3.4

0.5

8.7

8.7

8.7

15.1

6.4

15.1

22.4

7.3

4.4

4.4

4.4

9.3

4.9

Sunkist

15.8

19.9

4.1

4.5

Orange Fruit

22.3

27

4.7

5.4

0.01%

0.02%

0.03%

0.04%

0.05%

0.10%

Tropicana Twister

21.9

21

13.9

11.7

6.4

0.5

7.5

Graph

12.6

5.6

12.6

18.5

5.9

Analysis
In order to properly determine the concentration of Vitamin C of the orange juices
and of the orange fruit, a calibration curve must be drawn and the volume of
Vitamin C needed to decolourise the juices then can be used to measure the
concentration of Vitamin C in the juices. Ideally, a calibration curve should be
drawn with every concentration between a 0.01% interval. However, due to time
and other constraints, only a few concentrations were used. This calibration curve
graph shows us that as the concentration of Vitamin C increases, the volume of
Vitamin C needed to decolourise the DCPIP decreases. This is because as the
concentration of Vitamin C increases, there are more particles of Vitamin C in that
particular volume. As the number of particles has increased, there is a higher
chance of collisions between the Vitamin C particles and the orange juice particles,
and therefore, it is easier for them to react and thus the DCPIP turns colourless at a
quicker rate because neutralisation happens at a quicker rate. There is a difference
of -98% between the volume needed to decolourise the DCPIP at 0.01% and at
0.10%. According to the error bars on the graph (that shows the highest and the
lowest value obtained during the repeats of the experiment- thus highlighting that
as the average of all 3 repeats was plotted, the actual true value of that particular
experiment could have been in between the length of the error bars), the results are
quite reliable as the error bars seem to be quite low and small. The smaller the error
bar, the more reliable and accurate the results are because there is a higher chance
of the true value being closer to the average value, stressing that the values
obtained during the repeat were reliable and close to the true volume needed to
decolourise the DCPIP at that particular concentration. Besides that, even though
there seems to be a very large difference between the 0.01% and the 0.10%, it
must be remembered that the experiment wasnt measured in distanced values and
instead, was measured in 0.01% intervals from 0.01% to 0.05% and then it was
skipped to 0.10%.
According to the carton, Sunkist Orange juice had the component of 9mg/100ml of
Vitamin C and Tropicana Twister had the component of 14mg/100ml. It took 7.5 ml
of Tropicana Twister to decolourise the DCPIP and it took 4.5 ml to decolourise
Sunkist whereas it took 5.4 ml of orange juice from the orange fruit to decolourise
the DCPIP. This meant that Tropicana Twister had a concentration of 0.047% of
Vitamin C and Sunkist had 0.066% of Vitamin C and the orange fruit had 0.058%
concentration of Vitamin C in it.
{To calculate the concentration of Vitamin C in the orange juices provided is a
complicated method. First, we would have to find the number of moles of Vitamin C
used in the 0.01% experiment. This can be done through the method of using the
calculation moles=concentration x volume. After finding the moles, the mass can
be found by multiplying the moles by the formula mass of Vitamin C, which is 176.
After finding the mass, a ratio can be used to deduce the mass of Vitamin C in the
orange juices. By doing this, the mass of Vitamin C in Tropicana Twister can be

deduced to be 5.104mg/ml and the concentration of VitaminC in Sunkist is

4.4mg/ml}

Conclusion
Although fruit juice is a good source of vitamin C, it is not the healthiest way to
obtain the vitamin, since fruit juices are very high in sugar/simple carbohydrates.
Juices contain either added sugar or sugars that occur naturally in the fruit and
become highly concentrated in the juice. Eating a serving of fresh fruit is far
healthier because the effects of the natural fruit sugars are moderated by the fiber
in the fruit. Even though the percentage of Vitamin C in Tropicana Twister was
higher than the percentage of Vitamin C in the orange fruit, we do not know the
origins of Vitamin C in the Tropicana Twister, which could be dangerous to the body.

Systematic Errors
A systematic error is usually down to a variable that cannot be controlled which
affects the entire experiment. The results produced should still be reliable and and a
trend may be observable but it is only to a certain degree. An example of a
systematic error in this experiment could be the temperature change as the
temperature of the surrounding constantly changes, which could affect the results.
There could also be a fault in the burette that results in an incorrect reading of the
volume of the orange juice used in the experiment, although this would probably
produce a consistent level of error, shifting the results in the same way, every time.
The results are most likely to be inaccurate but they are still reliable.

Random Errors
A random error is a mistake in the method or malfunction in the equipment that
leads to an anomalous result. When a random error is spotted, it should be ignored
or the experiment should be repeated to produce a result that is more consistent
with the other results (this, however, has its disadvantages as the one anomalous
result could be the actual result). A mistake in this experiment could be due to a
wrong reading in the burette or the solution in the burette wasnt read at eye level,
which results in a wrong measurement of volume. The mistake could also be due to
there being an air bubble at the end of the burette which, again, shows us a wrong
measurement of the actual volume as the air bubble would add to the volume of the
solution at the start, and not be actually at the level 0. Another random error could
be the fact that we might have misjudged the end point as it is hard to locate the

final point exactly. However, again, this was hopefully eliminated as the experiment
was repeated.

Evaluation
Validity
Having kept all the other possible independent variables constant (such as the
temperature of the orange juice or the concentration of the DCPIP solution and the
same end point), and only changing the type of orange juice used, the results of the
experiment would be as reliable as possible. To ensure that the results are to be
reliable as possible, the orange juices were both refrigerated and taken out at the
same time and therefore, the temperature of the orange juices would have been
kept the same as possible and the Vitamin C content in the juices wouldnt be
reduced by much and we can measure the Vitamin C content of the juices as close
to the real value as possible as none of the juices had been heated.
The volume or the concentration of the DCPIP were kept the same as possible as
they could lead to inaccurate results as they can change our results because it
might take more orange juice to dilute a more concentrated DCPIP solution.
The results from the experiment were consistent with our hypothesis, as the bottle
juices had a lesser Vitamin C content than the actual orange.
As Vitamin C is a vitamin that is easily oxidisable, if the orange fruit was left out for
a certain period of time before using it, it might reduce the Vitamin C content in the
fruit as the Vitamin C gets oxidised by the oxygen in the air. Therefore, the orange
fruits must be immediately used after it is cut.

Precision
Ensure that the orange juice is added to the burette at eye level and any readings
are taken at eye level, to ensure that the volume measured is as precise as
possible. Also, make sure that the readings taken are recorded in a neat table and
can be read off and compared later to prevent any confusion. As we used a burette
in the experiment, the results were read of at a very precise level as the uncertainty
of our measurements were extremely small . The precision of our results could be
improved if we had used syringes to measure the volume of DCPIP rather than using
pipettes.
Accuracy
In this experiment, it is very important to control temperature as the Vitamin C
concentration in the orange juices could change if the temperature gets too hot as
Vitamin C is sensitive to temperature. It is also very important to measure the
equipment accurately and ensure that the reading is as accurate as possible.

As we used a burette in our experiment, the readings of allowed more accuracy as

each marker increases by 0.1 cm^3 rather than 0.5 cm^3. All of the readings were
read off at eye level and at the bottom of the meniscus. The Vitamin C juices were
added drop by drop and the solution was swirled to maximise the rate of reaction.
Also, even if precise equipments were used, the size or the volume of the drops
couldnt be controlled, which might mean that the size of the one drop could be
different to the size of another drop. However, this was overcome as the experiment
was repeated.
Reliability
When adding the orange juice to the burette, make sure that the orange juice is
filtered first to ensure that there is no pulp in the juice, which might block out the
opening of the burette and also give you a wrong reading of the volume of the
Orange juice added to the burette as the pulp could add on to the volume of the
orange juice in the burette. This could affect the reliability of our results.
The reliability of our results also depends on the amount of shaking of the tube as
we realised that we did not control that by much. The reason for this is that if we
were to shake the tube by a lot, more oxygen would be dissolved into the DCPIP,
which would restore the DCPIP tp blue.
The end point is also particularly hard to measure as we have to measure the exact
moment when the colour blue disappears, which was hard and we could have easily
missed the end point and kept titrating even when we have passed the end point.
We repeated our experiment 3 times for each Vitamin C solution and for each type
of orange juice to ensure that the results were reliable and we managed to notice
any outliers or anomalies as we took the average of our results. For the most of our
results, the range between the highest and lowest result was small, and therefore,
the results can be considered as reliable.

Bibliography
http://carbon.indstate.edu/inlow/LabManuals/Vitamin%20C.pdf
file:///C:/Users/ACER/Downloads/AS_biology_core_practical_summary%20(2).pdf
http://carbon.indstate.edu/inlow/LabManuals/Vitamin%20C.pdf
https://www.scribd.com/doc/165025653/Biology-Lab-Report-Vitamin-C-experiment
https://www.scribd.com/doc/165025653/Biology-Lab-Report-Vitamin-C-experiment
http://shinaebiochem.blogspot.com/2013/05/experiment-4-experiment-on-vitaminc.html
http://filestore.aqa.org.uk/subjects/AQA-HB13X-TN-JUN13.PDF