Aelfgar College

Biology AS

Investigating the vitamin C contents

of different sources
Anna Phizacklea
Introduction
Aim: to use dichlorophenolindolphenol (DCPIP) to determine the vitamin C contents of
different sources.

Hypothesis: The orange will contain the most vitamin C in mg/cm³.

Independent variable: The different sources used: orange juice, lime, lemon, orange.

Dependent variable: The volume of juice (attained from the source) required to
decolourise 2cm³ of DCPIP. This can be used to calcuate, when compared with the control
of 0.1% vitamin C solution, how much vitamin C the source contains in mg/cm³.

Control variable Possible effect Control measure

Age of the source
The amount of vitamin C in a
piece of fruit decreases over
time. This means if they are of
different ages, it will not give
accurate results.
Use fruit of the same age; fresh
is preferable, as vitamin C may
be lost at different rates.

Background information: Citrus fruits, such as lemons, contain vitamin C which prevents
the deficiency disease scurvy. Vitamin C is a reducing agent, and decolourises the blue
dye DCPIP. This reaction allows us to estimate the vitamin C content of different sources.

Equipment and materials

 Small beakers to collect volumes of juice (100cm³)
 Two large beakers (500cm³)
 Measuring cylinder (50cm³)
 Pipette or syringe to measure 2cm³ volume
 Pipette or syringe to accurately measure up to 1cm³
 Test tubes
 Spatula
 Distilled water (50cm³)
 0.1% vitamin C solution
 DCPIP

The sources used for this example:

 A lemon
 An orange
 A lime
 Shop-bought orange juice
Method
 Using a pipette or syringe, place exactly 2cm³ of DCPIP solution into a test tube
 Using a different graduated pipette or syringe, add the vitamin C solution, drop by
drop, to the DCPIP solution in the test tube. Shake the tube gently after the addition
of each drop and continue adding drops until the DCPIP solution is decolourised.
Record the exact volume of vitamin V solution added.
 Repeat this procedure. Average the results to obtain the mean volume of 0.1%
vitamin C which decolourises 2cm³ of DCPIP.
 Calculate the mass of vitamin C in this volume of vitamin C solution (it contains
0.001g vitamin C in 1cm³ water). This tells you how much vitamin C is required to
decolourise 2cm³ of DCPIP, which can be used to calculate the volume of vitamin C
in the different sources.
 First determine the volume of one of the sources, e.g. the lemon. Slowly submerge
it in a 500cm³ beaker of water, and use another 500cm³ beaker or container to
collect the water displaced. The volume of water displaced is the volume of the
lemon.
 Cut your lemon in half. Squeeze the juice of the lemon into a small beaker. Collect
some of the lemon juice using a clean graduated pipette or syringe.
 Repeat steps 1 and 2 using the juice attained from your source. Add the juice, drop
by drop, to 2cm³ of DCPIP, until it becomes decolourised.
 Repeat steps 5-7 using your different sources. Obviously in the case of orange
juice, you do not need to use displacement to measure the volume – go straight to
step 7.
Results

Raw data
Volume / cm³ required to decolourise 2cm³ DCPIP
Solution 1 2 3 4 Mean
0.1% vitamin C 1.5 2 1.5 1.6 1.65
Orange juice 4.5 4.5 4.3 4.4 4.43
Lime 6.5 7 7.4 7 6.98
Lemon 4 6.4 5.2 5.5 5.28
Orange 6 7.8 6.5 6.3 6.65

Calculated data
From these results, you can calculate the vitamin C content in mg/cm³ of each of the
sources, and using the volume of the source, the mg of vitamin C in each piece of fruit.
1cm³ of 0.1% vitamin C solution is 1mg vitamin C, therefore 1.65mg vitamin C
decolourises 2cm³ DCPIP.

Solution Mean volume mg required ÷ mg/cm³ mg/100cm³ Volume mg of

to decolourise volume of of fruit vitamin C
2cm³ DCPIP solution required piece/cm³ per piece
of fruit
0.1% 1.65 1.65 ÷1.65 1 100 – –
vitamin C
Orange 4.43 1.65 ÷ 4.43 0.37 37.3 – –
juice
Lime 6.98 1.65 ÷ 6.98 0.24 23.7 94 22.24
Lemon 5.28 1.65 ÷ 5.28 0.31 31.3 100 31.27
Orange 6.65 1.65 ÷ 6.65 0.25 24.8 180 44.66
Conclusion
As can be seen on the graph, orange juice contains the most vitamin C, at 0.37mg/cm³,
making it the best source of vitamin C. The second best source was the lemon, at
0.31mg/cm³. Contrasting with the hypothesis, the third best was the orange, at
0.25mg/cm³. The least vitamin C containing source was the lime, at 0.24mg/cm³. However,
if related to size, the orange contained 44.66mg of vitamin C in one piece of fruit,
compared to 31.27mg in a lemon, and 22.24mg in a lime, meaning the orange would be
the best source overall if a person were to choose one specific fruit to eat, as the amount
of vitamin C in orange juice would vary depending on the volume of juice.

Evaluation
There were numerous sources of error throughout the experiment. One error was the
argument of when the DCPIP had gone clear, as many people had different views of when
it was. Also, as the juices obtained and used were coloured, it was difficult to decide when
it had gone clear as it was tinted with the colour of the juice. Within each group, this was
avoided by having the same person decide when it was clear. However, there were
discrepancies between groups. This could be avoided by a pre-arranged definition of clear
from the whole class.
Another source of error was the pipettes used. Some of them were graduated with smaller
measures, and were therefore more accurate than others, so in some cases it was
uncertain exactly how much juice from a source had been added. This could be avoided
through everyone using the same kind of pipette for each purpose, preferably one with
small graduations for optimum accuracy. Another problem with the pipettes was that some
of the ones used did not hold the liquid in well, and a small amount escaped before
reaching the test tube. This could have had an effect in the case of measuring DCPIP, as
there would be less to be decolourised, tainting the results. This could be avoided by
ensuring the pipettes used were fully functional beforehand, and making sure the pipettes
still contained the correct amount before placing in the test tube.
An additional source of error could be the amount the test tube was shaken after each
drop of a vitamin C containing solution was added. Some people may have shaken it for
longer than others, which could have meant the vitamin C dispersed more or acted faster
than in other cases. This could be avoided by having a pre-set amount of time to shake for,
e.g. 20 seconds, and having the same person in each group do so each time.
A problem with the results obtained is that they are based on solely the juice taken from a
fruit. This ignores the other parts of the fruit, i.e. the rind and fleshy parts. This makes the
assumption about how much vitamin C is contained in a piece of fruit unreliable. To
improve this, the experiment could be tried by cutting up and processing a whole piece of
fruit, and adding the obtained liquid to the DCPIP. This would take into account the whole
of the fruit, although there may be further issues with viscosity and clearness.