Fletcher Sarip

th
May 11 2009
Introduction
Vitamin C is

really needed in our body. It has an important role in the synthesis of

neurotransmitter, which has big effect to the brain function and also may affect
the mood of a person. Our immune system also needs this vitamin, for it is also
an effective antioxidant which can protect the important molecules in our body,
such as protein, carbohydrates, lipids, and nucleic acids from the damage made

by reactive
oxygen species and free radicals. Vitamin C also helps the producing of Vitamin
E, which is also an antioxidant.

Our

body cannot produce Vitamin C by its own, and so we need to eat fruits and
other foods which contain this vitamin. Orange is the fruit which is well-known as
a Vitamin C container, and orange squeeze is popular and liked by most of the
people. They often prefer to drink the orange squeeze in cold temperature
because it is more refreshing than drinking it in normal temperature. However,
do you ever consider if the temperature will affect the amount of Vitamin C
contained in the orange squeeze? Will the cold or hot temperature decrease the
Vitamin C?

Lab Objective:
The lab objective is to find out about the effect of temperature towards
the amount of Vitamin C contained in an orange squeeze. We will try to find the
amount of Vitamin C in 3 orange squeezes which have the temperature of
normal temperature, hot temperature, and cold temperature. To find out the
amount of Vitamin C, which is also known as ascorbic acid, we will use
iodometric titration. This method uses the water-soluble characteristic of Vitamin
C which can be easily oxidized. We will use the iodine to oxidize the Vitamin C.
Before the Vitamin C is oxidized, we will dissolve starch into the orange squeeze
first. After that, we will do the titration of the iodine towards the orange squeeze.
The iodine will then reduce the Vitamin C in the orange squeeze until the whole
Vitamin C is reduced, and then the next drop of iodine will react with the starch
in the orange squeeze, which will form black color in the squeeze. The forming of
the black color is the sign of the end of titration. The volume of reacted iodine
from the beginning of the titration up to the forming of the black color can be
used to find the number of moles in the iodine. From the knowledge that the
density of iodine is 4.933 g/ml, we can find that the molarity of iodine is 0.03858
mol/ml (by dividing 4.933 g with the mass of 1 mol of iodine, which is 127 g, to
find that 4.933 g is 0.03858 mol of iodine), and by using this molarity, we can
find the number of moles of iodine in a certain volume by multiplying the
molarity with the reacted iodine volume. Since the ratio between iodine and
ascorbic acid (Vitamin C) is 1 to 1, the number of moles of the Vitamin C will be
equal with the number of moles of the iodine that we will find. We then can
multiply the number of moles of the ascorbic acid with the molar mass of
ascorbic acid (which is 176.1mg/mol) to find the mass of the Vitamin C.

Hypothesis:
My hypothesis is that the higher the temperature is, the less amount of
Vitamin C the orange squeeze will have, for I think the high temperature may kill
the vitamin. Besides that, high temperature may cause evaporation towards the
orange squeeze and the Vitamin C inside that may decrease the amount of the
orange water and also the amount of Vitamin C. Also, usually something related
to being burned will always turns into carbon, which means that if I increase the
temperature of the orange squeeze, some of the molecules inside including the
Vitamin C may start to turn into carbon as well.

Variables:
• Independent:
○ Temperature
 I will use the orange squeeze with different temperature. I will
make an orange squeeze first, and then I will set it in normal
temperature room (which is around 25 – 27oC), in cold
temperature (around 10oC), and in hot temperature (around
50oC).
• Dependent:
○ Amount of Vitamin C
 I will do the iodometric titration to find out the amount of
Vitamin C in the orange squeeze.

• Control:
○ Volume of orange squeeze
 The volume of the orange squeeze is set equal in all
temperature at 10 ml.
○ Freshness of the orange squeeze
 The orange is investigated right after the squeezing so that
all of them have the same level of freshness.
○ Amount of starch
 The amount of starch put into the orange squeeze in all of
the assigned temperature is equal.
Method

Materials:
1. Orange fruit = 12 fruits
2. Betadine (1% iodine)
3. Starch

Apparatuses:
1. Small beaker = 3 glasses
2. Orange squeezer =1
3. Beaker glass = 3 glasses
4. Stirring rod =1
5. Small spoon =1
6. Thermometer =1
7. Anti-heat gloves = 1 pair
8. Hot plate
9. Refrigerator
10.Pipette =3

Method:
1. First, squeeze 4 oranges to get the orange squeeze.
2. Put the orange squeeze into a glass beaker.
3. Measure the temperature of the orange squeeze using the thermometer
and take notes of the temperature.
4. Then use the pipette to put the orange squeeze into a small beaker for 10
ml.
5. Drop the starch 6 times to the orange squeeze by using the small spoon.
Each drop consists of the volume of starch at the tip of the small spoon.

Picture 1

6. Stir the orange squeeze with the stirring rod until the starch is dissolved.
7. Give 1 drop of Betadine and shake the orange squeeze. DROP THE
BETADINE STRAIGHT TO THE ORANGE SQUEEZE. DON’T LET THE
DROP OF THE BETADINE TOUCHES THE WALL OF THE SMALL
BEAKER, FOR IT MAY DECREASE THE VOLUME OF BETADINE IN ONE
DROP AND MAY COMPLICATE THE FURTHER CALCULATING.
8. If the orange squeeze doesn’t change into black color, keep dropping the
Betadine to the orange squeeze and remember to shake the orange
squeeze for each drop of Betadine.
9. ALSO REMEMBER TO COUNT HOW MANY DROPS OF BETADINE YOU
HAVE PUT.
10.Do it continuously until the orange squeeze’s color changes to black when
you shake the small beaker.
 Before: After:

Picture 2 Picture 3

11.One drop of Betadine consists of 1/20 ml. Since the amount of iodine in
one drop of Betadine is 1%, we can find the volume of reacted iodine by
doing the calculation:

1%×How many drops of Betadine×120ml

12. Now, multiply the volume of reacted iodine with the molarity of iodine to
find the number of moles of iodine:

total volume of iodine×0.03858 mol/ml

13.Then multiply the number of moles of iodine (which will be equal to the
number of moles of ascorbic acid (Vitamin C)), with the molar mass of
ascorbic acid (176.1 mg/mol) to find the mass of Vitamin C.

number of moles of iodine×176.1 mg/mol

14.Repeat this experiment for 3 times.

15.Take notes of the data in this table:

Temperat Tri Drops Total Volume of Total Number of Moles of Mass of Vitamin C
ure al of Dropped Volume Iodine number of moles
(oC) Betadi Betadine of total volume of of
ne drops of Iodine iodine×0.03858 iodine×176.1
(drops) betadine×120ml Droppe mol/ml mg/mol
d
(1%×
Total
Volume
of
Droppe
d
Betadin
e) (ml)

13.After that, squeeze 4 oranges again and put the orange squeeze on
another beaker glass.
14.Increase the temperature of the orange squeeze in the beaker glass by
using the hot plate.
15.While heating the orange squeeze, put the thermometer in the beaker
glass to be able to see the temperature.
16.Wait until the temperature of the orange squeeze is 70oC, then turn off the
hot plate quickly, for the temperature may still increase even the tool is
turned off, so that after turning off the tool, the temperature of the orange
squeeze will still increase only up to 75oC. In this case, the temperature
that should be written in the note is 75oC.
17.Use the anti-heat gloves to take the beaker glass from the hot plate.
18.Repeat step 4 – 14 (use new pipette and small beaker glass) and take
notes of the data in the table above.
19.After that, squeeze 4 oranges again and put the orange squeeze in the
other beaker glass.
20.Put the orange squeeze in the refrigerator and check the temperature of
the orange squeeze frequently using the thermometer until the
temperature is 10oC.
21.Take out the orange squeeze from the refrigerator and repeat step 4 – 14.
Take notes of the data in the table above.
 Result

This is the result of the experiment:

Temperatur Trial Drops Total Volume Total Number of Mass of Vitamin C
e of of Dropped Volume of Moles of (number of moles of
(oC) Betadin Betadine Iodine Iodine iodine x 176.1
e (drops of Dropped (total mg/mol)
(drops) betadine x (1%× volume of
1/20 ml) Total iodine x
Volume of 0.03858
Dropped mol/ml)
Betadine
(ml))
0.006 ml x 0.00023148 mol x
1%×35 0.03858 176.1 mg/mol =
12×120ml=12
24.7 1 12 ml=0.00 mol/ml = 0.0408 mg
20ml=35ml
6 ml 0.00023148
mol
0.006 ml x 0.00023148 mol x
1%×35 0.03858 176.1 mg/mol =
12×120ml=12
2 12 ml=0.00 mol/ml = 0.0408 mg
20ml=35ml
6 ml 0.00023148
mol
0.006 ml x 0.00023148 mol x
1%×35 0.03858 176.1 mg/mol =
12×120ml=12
3 12 ml=0.00 mol/ml = 0.0408 mg
20ml=35ml
6 ml 0.00023148
mol

0.006 ml x 0.00023148 mol x

1%×35 0.03858 176.1 mg/mol =
12×120ml=12
75 1 12 ml=0.00 mol/ml = 0.0408 mg
20ml=35ml
6 ml 0.00023148
mol
0.006 ml x 0.00023148 mol x
1%×35 0.03858 176.1 mg/mol =
12×120ml=12
2 12 ml=0.00 mol/ml = 0.0408 mg
20ml=35ml
6 ml 0.00023148
mol
0.006 ml x 0.00023148 mol x
1%×35 0.03858 176.1 mg/mol =
12×120ml=12
3 12 ml=0.00 mol/ml = 0.0408 mg
20ml=35ml
6 ml 0.00023148
mol

0.006 ml x 0.00023148 mol x

1%×35 0.03858 176.1 mg/mol =
12×120ml=12
10 1 12 ml=0.00 mol/ml = 0.0408 mg
20ml=35ml
6 ml 0.00023148
mol
2 12 12×120ml=12 1%×35 0.006 ml x 0.00023148 mol x
20ml=35ml ml=0.00 0.03858 176.1 mg/mol =
6 ml mol/ml = 0.0408 mg
0.00023148
 mol

0.006 ml x 0.00023148 mol x

1%×35 0.03858 176.1 mg/mol =
12×120ml=12
3 12 ml=0.00 mol/ml = 0.0408 mg
20ml=35ml
6 ml 0.00023148
mol

Table 1
So, the amount of Vitamin C in each temperature is:
Figure 1
Discussion
The result of the experiment shows that my hypothesis is wrong. In Table
1, it is shown that 10 ml of 24.7oC orange squeeze needs 12 drops of Betadine
until the color changes to black, while the other orange squeezes with the same
volume and the temperature of 75oC and 10oC also need the same drops of
Betadine to react. One drop consists of 1/20 ml Betadine and contains 1% of
iodine, which means that only 1% iodine of 1/20 ml Betadine that reacts with the
ascorbic acid in the orange squeeze for each drop. So, in 12 drops of Betadine,
the total volume of iodine reacted with the ascorbic acid is 1% of 12 times 1/20
ml Betadine, which is 0.006 ml. With the molarity of iodine which is 0.03858
mol/ml, we can find the total number of moles of the iodine, which is 0.00023148
moles. The number of moles of iodine is equal to the number of moles of
ascorbic acid (Vitamin C) since the ratio between iodine and ascorbic acid is 1 to
1, and so with this number of moles, we can find the mass of Vitamin C by
multiplying the number of moles with the molar mass of Vitamin C (176.1
mg/mol), which then the result is 0.0408 mg. Since the other orange squeezes
with different temperature also need the same drops of Betadine, it means that
all of the orange squeezes have the same amount of Vitamin C, as shown in the
Figure 1. This shows that the temperature doesn’t affect the amount of Vitamin C
and doesn’t even kill nor turn the Vitamin C into carbon as I think in my
hypothesis.
In raising the temperature of the orange squeeze, I am using a tool which
raises the temperature of the air around that then will also raise the temperature
of the orange squeeze instead of burning it directly by using fire. I think by using
this method, the temperature will not affect the amount of Vitamin C in the
orange squeeze since the Vitamin C is not directly exposed to heat and the
temperature is only affected by the hot temperature of the air, which also means
that the Vitamin C is not directly burned and there is no reaction of
carbonization. This is why the amount of Vitamin C stays the same even though
the temperature of the orange squeezes is raised or lowered.
The data collected is not really reliable. The volume of 1 drop of Betadine
may not be exactly 1/20 ml, for the size of each drop may accidentally becomes
bigger or smaller slightly according to how hard we press the bottle of the
Betadine and to any other conditions. This means that there may be accidentally
small difference of reacted iodine and the amount of Vitamin C between the
orange squeezes with the different temperature that may not be recognized if
we use the idea of 1 drop equals to 1/20 ml as the basic idea. It may be that the
amount of Vitamin C in the hot orange squeeze is slightly smaller or bigger than
in the normal orange squeeze that we don’t recognize by using this kind of idea.
Conclusion
From this investigation, we can learn that the temperature doesn’t have
any effect towards the amount of Vitamin C in the orange squeeze. It is a direct
exposure of heat, such as fire, towards the orange squeeze that may burn and
decrease the Vitamin C, while in this experiment the temperature is raised by
the hot air around the orange squeeze produced by the tool which doesn’t give
any effect to the Vitamin C inside. Besides that, we can also learn that Vitamin C
can be reduced if it is reacted with iodine and that the reaction between iodine
and starch will form black color. By looking at the data above, we can see that 10
ml orange squeeze contains about 0.0408 mg Vitamin C. From this data, we can
conclude that in 1 ml of orange squeeze there is 0.00408 mg of Vitamin C. From
this data, we can know how much orange squeeze we should drink each day to
meet the RDA recommendation for Vitamin C.
While doing the lab experiment, I made some little mistakes that make the
information becomes unreliable. Some drops of Betadine that I dropped into the
small beaker touched the wall of the glass, which then as it flowed downward
through the wall some of it was left behind for it stuck on the wall of the glass,
and when it touched the orange squeeze the volume of the Betadine would not
be 1/20 anymore. In facing this problem, I didn’t take it as 1 count of drop and
just ignored it. This makes the data in the table doesn’t exactly show the exact
volume of Betadine and iodine reacted with the orange squeeze because I didn’t
count the error that happened in the experiment which may give additional
volume of reacted iodine and Betadine towards the orange squeeze.
The method that I used may not be able to control the variables well. The
amount of starch put into the orange squeeze may not equal between every
orange squeeze because I used the tip of the small spoon as the measurement of
the volume of the starch which is not a reliable measurement. Sometimes the
starch may be more or less if we take it by using the tip of the small spoon. This
difference of the amount of starch dissolved in the orange squeeze may affect
the reaction result. Besides this problem, the freshness of the orange squeeze
cannot be controlled too, for the heating and refrigeration of the orange squeeze
will need some time before they are investigated, while the orange squeeze of
the normal temperature will be directly investigated right after the squeezing.
This means that the freshness of orange squeeze cannot be controlled for the
orange squeeze of the normal temperature will have more freshness than the
orange squeezes with the cold and hot temperature while they are investigated.
For the next time, I suggest that the method should be changed a little bit.
For the amount of starch put into the orange squeeze, we should measure it
based on the mass that can be measured by using a reliable tool such as a scale
which has constant measuring so that we can give the exact same amount of
starch to every orange squeezes by looking at the mass. Also, to control the
freshness of the orange squeeze, I think we should set the same amount of time
before the orange squeezes are investigated, such as to wait for 10 minutes (for
the normal temperature), heating for 10 minutes (for the hot temperature), or
refrigerating the orange squeeze for 10 minutes (for the cold temperature) after
squeezing the oranges before investigating the amount of Vitamin C. By doing
this, the freshness of every orange squeeze may be the same.
This lab still only focuses on the effect of orange squeeze temperature
towards the amount of Vitamin C inside. For further inquiry, I suggest to
investigate the effect of temperature towards the other kinds of vitamin, for
there may be some vitamins which are and aren’t affected by the temperature.
Besides that, the effect of the time of heating by using fire towards the amount
of vitamin C and the other vitamins may also be investigated, because direct
exposure towards heat is the biggest factor of vitamin loss. Also, we can
investigate the effect of temperature towards the speed of vitamin loss after the
fruit or fruit juice is left for several hours, for it may be useful in finding a perfect
way to store fruits or fruit juices without losing a lot of vitamin inside.
One World
Our body needs Vitamin C a lot, especially in defending ourselves from the
invasion of viruses, for Vitamin C is an antioxidant. Vitamin C can protect our
important molecules in our body such as protein, carbohydrates, lipids, and
nucleic acids from damaging made by reactive oxygen species and free radicals.
It also gives some help in producing Vitamin E which is also an antioxidant.
Getting enough Vitamin C may protect our body from some chronic
diseases. Consuming Vitamin C for 100 mg/day may prevent cardiovascular
disease for nonsmoking men and women. It is found that people who consume
Vitamin C more than 50 mg/day have lower risk of death from cardiovascular
disease, which is 42% lower in men and 25% lower in women. Besides
cardiovascular disease, Vitamin C can also reduce the risk of stroke, cancer,
cataracts, and lead toxicity. Other than preventing, treating diseases is also the
thing that Vitamin C can do. Vasodilatation, hypertension, cancer, diabetes
mellitus, and common cold are the diseases which can be treated by consuming
Vitamin C to increase the survival rate and decrease the chance that the disease
becomes more serious.
Since our body cannot produce Vitamin C by its own, we need to eat fruits
and foods which contain Vitamin C, and one of the popular fruits which contain
Vitamin C is orange. People often like to squeeze the oranges because it will be
easier to consume. Orange squeeze is also popular in the restaurants and every
time you go to any eating places, you will find orange squeeze in the menu book.
Most people like to drink the orange squeeze in cold temperature. By knowing
that the temperature will not affect the amount of Vitamin C in the orange
squeeze, we don’t need to worry about the vitamin loss. Also, there may be
some cases that people get sick and need to get a lot of Vitamin C while they
should not drink any cold drinks such as in fever, cough, or other diseases. Then,
by knowing that the temperature doesn’t have any effect towards the amount of
Vitamin C, they can just drink the orange squeeze in hot temperature if they
want to without worrying about the vitamin loss.
Besides that, companies also don’t have to worry about storing the orange
juices that they have produced in the cold freezer since the temperature doesn’t
have any effect towards the Vitamin C inside the orange juices. They can just
store it in a normal container without worrying of losing the Vitamin C inside
instead of using freezer as the container that may produce CFC which is one
third of the cause of greenhouse effect in our world. Also, by using a normal
container, the companies can spend less money in paying the cost of the storing
of their products, which may be higher if they use the freezer as the storing
place.
However, the vitamin C may keep the same only if the temperature is
affected by the temperature of the air. If the orange juice is directly exposed to
the heat, such as fire or sun, the amount of Vitamin C may decrease. This means
that this theory cannot be applied if a company wants to transport their orange
juice products from one place to another by using a truck, for the orange juices
may be exposed to sunlight directly which may decrease the vitamin C inside.
This means that the company should provide a fully covered location for the
transportation of the orange juices.
 References

Boyter, Henry. "Chemistry (including Biochemistry): Vitamin C Research

Project, polyethylene wax, mole ratio." Expert Archive Questions. 4

Dec. 2004. 11 May 2009 <http://en.allexperts.com/q/Chemistry-

including-Biochemistry-1355/Vitamin-C-Research-Project.htm>.

Church, Steve. "Investigation Report Format." Investigation Report Format

(2009).

Higdon, Jane. Linus Pauling Institute at Oregon State University. 17 Oct.

2008. Linus Pauling. 11 May 2009

<http://lpi.oregonstate.edu/infocenter/vitamins/vitaminC/>.

"Iodine definition | Dictionary.com." Dictionary.com. 2006. Houghton

Mifflin Company. 11 May 2009

<http://dictionary.reference.com/browse/iodine>.

"What is Greenhouse Effect ?" Greenhouse Effect. 4 May 2003. Hongkong

Observatory. 11 May 2009

<http://www.weather.gov.hk/wxinfo/climat/greenhs/e_grnhse.htm>.