Completepaper 1

Eskuri Rupert Tablada
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21 May 2015
Introduction
Many people would associate the word scurvy with pirates, but little do
they know scurvy is also a disease. This disease was popular with sailors
between the sixteenth and eighteenth centuries who navigated through long
voyages. These sailors could have prevented scurvy and death from scurvy by
consuming more vitamin C. Although scurvy is rare today the elderly, alcoholics,
or those whose diet does not contain fresh fruits or vegetables can still be
diagnosed with it. Adults diagnosed with scurvy may suffer from symptoms such
as fatigue, feeling out of sorts, pain in your limbs - particularly your legs, and
the appearance of small red-blue spots on your skin which can merge to form
larger marks. Following those symptoms are more severe ones such as swollen
gums, severe joint pain, shortness of breath, redness and swelling in recently
healed wounds, and easily bruised skin. Infants diagnosed with scurvy may
suffer from symptoms such as lack of appetite, irritability, poor weight gain,
diarrhea, and high temperature. If it progresses additional symptoms are pain
and tenderness in the legs, the appearance of reddish-bluish spots on the skin,
and the eyes bulging outwards (Scurvy - Symptoms"). Scurvy can also lead to
future problems: anemia, debility, exhaustion, ulceration of the gums, and loss of
teeth (Crosta). This can be avoided by, of course, consuming more vitamin C.
Today, many people obtain their daily vitamin C regimen through juices such as
orange juice. Some people make their juice fresh at home by squeezing the juice
out of oranges, but the problem with this is that it is time consuming. People with

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busy lifestyles do not have time to sit down and squeeze oranges, so they buy
juice from the store. The issue with the store bought juice is that it is processed
so the juice is not fresh, and vitamin C deteriorates over time. If people are
getting their vitamin C through the store bought juice, they might no get enough
of the vitamin even if they believe they are. This can cause a disease caused by
vitamin C deficiency such as scurvy.
The purpose and objective of this experiment was to find out if there was
significantly more vitamin C in freshly squeezed organic orange juice or in store
bought Minute Maid inorganic orange juice. Oranges were chosen as the type of
juice concentration because it tends to be the most commonly thought of as the
fruit with the most vitamin C content in the juice. To determine which juice
contained more vitamin C titrations were done on the juices to determine the
molarity - or concentration - of vitamin C in the juice. Hydrochloric acid and 2%
starch solution were used as catalysts in the orange concentrations to aid in
finding the difference in vitamin C in both the organic and inorganic orange juice
concentrations. Iodine was then added into each trial to test the molarity for the
concentrations.
After the data was collected boxplots and dot plots were observed, and
the average concentrations of the juices were calculated to determine if there
was a difference in the concentrations. Once it was determined that there was a
difference, a t-test was performed on the data to determine if it was statistically
significant. By testing the difference of organic and inorganic orange juice
concentrations it was hoped to find the most efficient way of consuming the daily
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amount of vitamin C to reduce the possibility of vitamin C deficiency such as

scurvy. It was found that there is indeed more vitamin C in freshly squeezed
orange juice than there is in store bought orange juice.
Review of Literature
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The purpose of this experiment was to determine the vitamin C content in

freshly squeezed organic and Minute Maid processed inorganic orange juice
concentrate by a titration.
Orange juice is beverage that is commonly found in many households
that contains vitamin C. Despite many common beliefs orange juice is not as
healthy as it is made out to be. Ninety eight percent of orange juice sold in North
America is pasteurized (Dishaw). Pasteurization is the process of heating a liquid
to extremely high temperatures to remove the threat of diseases and pathogens
such as Escherichia coli and salmonella, but in the process of pasteurization
many nutrients, such as vitamin C, are destroyed leaving just sugar and water.
According to the author of the book Squeezed, Alissa Hamilton exposes that
orange juice is stored up to a year in tanks, and stripped of oxygen and flavor
(Hamilton). After this process is done, the orange juice is reflavored.. After the
juice is squeezed and stored in gigantic tanks, they start removing oxygen by
pasteurizing the juice. Pasteurization is a process where the juice is heated to
high temperatures to remove the pathogens and bacteria (Pasteurization).
When the orange juice is pasteurized, it takes away the flavoring, nutrients, and
preserves the juice to keep it from spoiling fast. In order to have orange juice
actually taste like oranges, drink companies hire flavor companies to create
"flavor packs" to make juice taste like orange juice again ("Dirty Little Secret:
Orange Juice Is Artificially Flavored to Taste Like Oranges").

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Conducting a titration lab on vitamin C in orange juice was chosen to find

the difference on how much vitamin C is actually in organic and inorganic juices,
and the differences between them. Vitamin C, also known as ascorbic acid, is
naturally in citric acid and in some fruits and vegetables. Vitamin C can be
oxidized easily; therefore, if an open orange was left out and exposed to the air,
the vitamin C levels will reduce (Vitamin C Content of Fruit Juice). The chemical
equation for vitamin C is C6H8O6.
Figure 1. The structure of vitamin C

Figure 1 shows the structure of vitamin C.
From the experiment (Vitamin C Content of Fruit Juice), the lab asked to
find the amount of vitamin C in one juice sample. In this experiment, orange juice
was the juice chosen, and the difference in vitamin C content with organic and
inorganic juices was found. That was the difference between the two labs, and
that there would be more than two trials. The experiment that was used to
determine
A titration is a procedure used to determine the concentration of an acid or
a base in a solution by adding an acid or base from a known concentration, a
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standard solution. Most titrations are done using a buret and a flask and
determining the volume of the known titrant from before and after the titration
("ChemLab - Techniques - Titration."). In the experiment being tested Iodine, the
known titrant, will be added to organic orange juice and inorganic orange juice
until the amount of iodine and the amount of vitamin C, the unknown titrant, in the
orange juice is exactly the same. When that occurs, it is called the endpoint of
the titration. The endpoint has been reached when a chemical called the indicator
- that is added to the orange juice - produces a permanent brown/black or
blue/black color ("Vitamin C In Fruit Skins."). The color will be considered
permanent when it lasts for longer than ten seconds, even when the solution is
stirred. If it is thought that the endpoint might have been reached, close the
stopcock and record the volume reading then add another drop of iodine, since
sometimes it may be easier to tell when the endpoint has been exceeded
("ChemLab - Techniques - Titration."). The endpoint of a titration can be shows by
a graph called a Titration Curve which is seen in figure 4 below. Iodine is used
because as it is added the ascorbic acid is oxidized to dehydroascorbic acid,
C6H6O6, and the iodine is reduced to iodide ions. After the vitamin C is oxidized,
or is chemically combined with oxygen, and turned into dehydroascorbic acid, the
leftover iodine reacts with the indicator forming the brown/black or blue/black
color (Science, College Of.). The chemical formula is
C6 H 8 O6+ I 2 2 I +C6 H 6 O6

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where the first C6 H 8 O6 is ascorbic acid,
I2
is iodine,
2 I is the two
iodide ions which is the color of the orange juice, and the second
C6 H 6 O6 is
the dehydroascorbic acid which turns the blue/black color.
Figure 2. Titration Curve

Figure 2 shows an example of a standard titration curve. As shown in the
example the equivalence point is found when the acid and base are equal
When the endpoint of the titration is reached the molarity of
ascorbic acid - or vitamin C - can be found. To do this the volume of the iodine
used, the volume of orange juice used, and the molarity of the iodine is used to
determine the molarity of the ascorbic acid. To determine the molarity of the
ascorbic acid in the orange juice the formula
M a V a=M b V b
where
M a is the molarity of the acid,
M b is the molarity of the base, and
Va
is the volume of the acid,
V b is the volume of the base ("Acid-
Base Titrations: Chapter 24."). To do these calculations however, make sure that
the chemical equation being used is balanced. Figure 3 below shows an example
of this equation along with an example of a balanced chemical equation.
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If 5.0 mL of 0.50 M NaOH is used to neutralize 10.0 mL of HC 2H3O2, what is the

molarity of the acid solution? The chemical formula for this problem is shown
below.
H C 2 H 3 O2 + NaOH NaC 2 H 3 O2+ H 2 O
M a V a=M b V b
M a 10=0.50 5
M a=
0.50 5
10
M a=.25
Figure 3. Example

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Problem Statement
Problem Statement:
Will organic or inorganic orange juice have a greater concentration of
vitamin C?
Hypothesis:
It is hypothesized that the organic orange juice will have 40% more
vitamin C than the inorganic orange juice.
Data:
The independent variable is the type of orange juice concentrate (organic
or inorganic). The organic orange juice will be freshly squeezed orange juice and
the inorganic will be Minute Maid orange juice. The dependent variable is the
concentration of vitamin C in the juice measured in moles divided by liters. The
test that will be used is a two sample t test.

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Experimental Design
Materials:
(2) Buret (50mL)
Ring Stand
Funnel
Beaker (250 mL)
10 mL Graduated Cylinder
(2)Graduated Cylinder (20.0 mL)
0.01M Iodine, I2
Distilled Water (2400mL)
Fresh Squeezed Organic Orange Juice(1200mL)
Minute Maid Orange Juice (1200mL)
2% Starch Solution
3.0 M HCl (Hydrochloric Acid)
Procedure:
Safety note:
1. Obtain safety goggles as chemicals can irritate the eye.
Preparation of the Burette:
2. Rinse the burette thoroughly with tap water to avoid any previous residue.
3. Open the stopcock to allow the rinse water to clean the tip of the burette as well.
4. Using a funnel, pour approximately 3 mL of 0.010 M iodine solution (the
preparation of the Iodine is shown in Appendix D on page #) into the burette.
With the stopcock closed, hold the burette in a nearly horizontal position and roll
the solution around to wet the entire inner surface of the burette.
5. Drain the iodine out of the burette by opening the stopcock. The reason for doing
this was to use a little portion of iodine to clear out any remaining water which
would otherwise tamper with the iodine solution and the results.
6. Close the stopcock. Clamp the burette to a ring stand on the lab bench using a
burette clamp.
7. Use a funnel to add approximately 40 mL of 0.010 M iodine to the burette.
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8. Fill the tip of the buret with iodine and place a clean beaker underneath the
burette to catch any liquid, then open the stopcock to drain the burette.
9. Close the stopcock again, and examine the tip of the burette to see if it is
completely filled with liquid or if there are some air bubbles trapped inside.
Titration of the Vitamin C in a Fruit Juice Sample
10. Use the random integer function on the Ti- Nspire calculator or on any
randomization program found on the internet to randomize whether the organic
orange juice or the inorganic orange juice will be titrated first (see Appendix B on
page #).
11. Clean the 250 mL flask, as the titration process will take place in that flask.
12. Use a 20.0 mL graduated cylinder to obtain 20.0 mL of juice and empty the
graduated cylinder of orange juice into the 250 mL flask.
13. Add 40 mL of distilled water to the orange juice concentration in the flask. Then
add 5 drops of 3.0 M HCl (hydrochloric acid) as a catalyst, and 10 drops of 2%
starch solution to serve as the indicator (the preparation of the starch solution is
shown in Appendix E on page #).
14. Place a white piece of paper under the flask to aid in distinguishing a color
change then begin the titration by draining about 1 mL of iodine out of the burette
into the flask containing the orange juice concentrate sample.
15. Stop and mix the solution.
16. Continue to slowly add iodine in this manner while still mixing the solution.
17. The process will be completed when there is enough iodine added that the dark
black color will appear and remained for at least 10 seconds without fading away
and remain permanent even after swirling the flask.
18. Complete the titration by slowly adding more iodine while mixing the solution
vigorously. When the endpoint is close, the iodine should be added very slowly,
only a drop at a time, so as not to go past the endpoint.
19. When the endpoint has been reached, take a final volume reading from the
burette and record data.
20. Discard the contents of the 250 mL flask down the drain and rinse the flask clean.
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21. Repeat steps 1-18 for the other orange juice concentrate.
22. When both titrations are finished, discard any unused iodine solution from the
buret down the drain and rinse the burette thoroughly with tap water followed by
distilled water.
23. Calculate the concentration of vitamin C to the ten thousands place in moles to
liters. As shown by the example in Appendix A.
Figure 4. Burette
Figure 4 shows a diagram of a burette
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Data and Observations

Table 1. Inorganic Orange Juice Observations/Comments
Trial
Inorganic Orange Juice Observations/Comments
The iodine was made 14/4/15. Everything was shaken and cleaned before
1
using. The beakers were mixed the whole time the titration was being
performed. Researcher 1 watched and shook the inorganic.
2
3
4
5
performed. Researcher 1 watched and shook the organic.
6
performed. Researcher 2 watched and shook the inorganic. The inorganic
ended up being very dark, and almost brown.
7
using.. The beakers were mixed the whole time the titration was being
8
9
10
11
The iodine was made 21/4/15 before the trial. Everything was shaken and
cleaned before using. The beakers were mixed the whole time the titration
was being performed. Researcher 2 watched and shook the inorganic.
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12
Trial
13
14
15
16
17
18
19
using. New Hydrochloric acid was made before the inorganic titration. The
beakers were mixed the whole time the titration was being performed.
Researcher 3 watched and shook the inorganic.
using. New Hydrochloric acid was made before the inorganic titration. The
beakers were mixed the whole time the titration was being performed.
Researcher 1 watched and shook the inorganic.
using. New Hydrochloric acid was made before the titrations. The beakers
were mixed the whole time the titration was being performed. Researcher 3
watched and shook the inorganic.
using. New Hydrochloric acid was made before the titrations. The beakers
watched and shook the inorganic.
20
using. New Hydrochloric acid was made 22/4/15. The beakers were mixed
the whole time the titration was being performed. Researcher 1 watched and
shook the inorganic.
21
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22
Trial
23
24
25
26
27
28
29
30

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Table 1 shows the observations made during the thirty inorganic juice
trials. There were three separate iodine solutions used throughout the trials and
the Hydrochloric acid was remade once. The iodine made on 21/4/15 originally
only had half of the amount of iodine needed, but the data was divided by two to
avoid any misinterpretations.
Table 2. Organic Orange Juice Observations/Comments
Trial
1
Organic Orange Juice Observations/Comments

The iodine was made 14/4/15, and the organic juice was made 10 min before
trials started. Everything was shaken and cleaned before using. The beakers
watched and shook the organic.
The iodine was made 14/4/15, and the organic juice was made yesterday.
Everything was shaken and cleaned before using. The beakers were mixed
shook the organic.
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Trial
10
11
12
13
14
15
16
The iodine was made 14/4/15, and the organic juice was made yesterday.
Everything was shaken and cleaned before using. The beakers were mixed
shook the organic.
The iodine was made 14/4/15, and the organic juice was made today right
before trials. Everything was shaken and cleaned before using. The beakers
The iodine was made today before the trial, and the organic juice was made
today right before trials. Everything was shaken and cleaned before using.
The beakers were mixed the whole time the titration was being performed.
Researcher 1 watched and shook the organic.
before trials. Everything was shaken and cleaned before using. New
Hydrochloric acid was made before the inorganic titration. The beakers were
mixed the whole time the titration was being performed. Researcher 2
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17
18
Trial
19
20
21
22
23
24
Hydrochloric acid was made before the inorganic titration. The beakers were
mixed the whole time the titration was being performed. Researcher 3
Hydrochloric acid was made before the titrations. The beakers were mixed
shook the organic.
Hydrochloric acid was made before the titrations. The beakers were mixed
shook the organic.
Hydrochloric acid was made 22/4/15. The beakers were mixed the whole
time the titration was being performed. Researcher 1 watched and shook the
organic.
organic.
organic.
organic.
organic.
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25
26
organic.
organic.
Trial
27
28
29
30

organic.
organic.
organic.
organic.
Table 2 shows the observations made during the thirty inorganic juice
trials. There were three separate iodine solutions used throughout the trials, the
Hydrochloric acid was remade once, and the juice was squeezed nearly every
day. The iodine made on 21/4/15 originally only had half of the amount of iodine
needed, but the data was divided by two to avoid any misinterpretations.
Table 3. Inorganic Orange Juice Data

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Trial
Inorganic Orange Juice

Volume
Molar Volume
Moles of
Difference Solution
of
Vitamin
of Iodine
of
Juice
C
(mL)
Iodine
(mL)
1
5.9 1.0E-02
60 9.8E-04
2
5.8 1.0E-02
60 9.7E-04
3
6.9 1.0E-02
60 1.2E-03
Trial
Inorganic Orange Juice
4
5.9 1.0E-02
60 9.8E-04
5
6.6 1.0E-02
60 1.1E-03
6
7.5 1.0E-02
60 1.3E-03
7
6.8 1.0E-02
60 1.1E-03
8
6.0 1.0E-02
60 1.0E-03
9
7.4 1.0E-02
60 1.2E-03
10
5.8 1.0E-02
60 9.7E-04
11
6.8 1.0E-02
60 1.1E-03
12
7.0 5.6E-03
60 6.5E-04
13
7.2 5.6E-03
60 6.7E-04
14
8.0 5.6E-03
60 7.5E-04
15
7.0 5.6E-03
60 6.5E-04
16
6.6 5.6E-03
60 6.2E-04
17
7.5 5.6E-03
60 7.0E-04
18
7.4 5.6E-03
60 6.9E-04
19
7.5 5.6E-03
60 7.0E-04
20
6.6 1.0E-02
60 1.1E-03
21
6.4 1.0E-02
60 1.1E-03
22
7.1 1.0E-02
60 1.2E-03
23
5.3 1.0E-02
60 8.8E-04
24
4.9 1.0E-02
60 8.2E-04
25
4.2 1.0E-02
60 7.0E-04
26
4.7 1.0E-02
60 7.8E-04
27
4.6 1.0E-02
60 7.7E-04
28
5.9 1.0E-02
60 9.8E-04
29
4.8 1.0E-02
60 8.0E-04
30
5.7 1.0E-02
60 9.5E-04
Average
6.3
NA
NA 9.1E-04
Table 3 shows the data collected during the inorganic trials. It also shows
the averages for the moles of vitamin C and the difference of iodine. It says NA,
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or not applicable, for the averages of the molar solution of iodine and the volume
of juice because they are constants.
Table 4.
Trial
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
Organic Orange Juice

Volume
Molar Volume
Difference Solution
of
of Iodine
of
Juice
(mL)
Iodine
(mL)
7.4 1.0E-02
60
7.5 1.0E-02
60
8.0 1.0E-02
60
7.5 1.0E-02
60
8.0 1.0E-02
60
7.0 1.0E-02
60
7.4 1.0E-02
60
6.2 1.0E-02
60
6.8 1.0E-02
60
6.9 1.0E-02
60
9.7 1.0E-02
60
9.6 1.0E-02
60
12.0 1.0E-02
60
12.9 1.0E-02
60
12.9 1.0E-02
60
13.2 1.0E-02
60
12.2 1.0E-02
60
13.3 1.0E-02
60
13.2 1.0E-02
60
12.6 1.0E-02
60
12.0 1.0E-02
60
12.6 1.0E-02
60
12.1 1.0E-02
60
7.8 1.0E-02
60
8.4 1.0E-02
60
8.7 1.0E-02
60
8.5 1.0E-02
60
21
Moles of
Vitamin
C
1.2E-03
1.3E-03
1.3E-03
1.3E-03
1.3E-03
1.2E-03
1.2E-03
1.0E-03
1.1E-03
1.2E-03
1.6E-03
1.6E-03
2.0E-03
2.2E-03
2.2E-03
2.2E-03
2.0E-03
2.2E-03
2.2E-03
2.1E-03
2.0E-03
2.1E-03
2.0E-03
1.3E-03
1.4E-03
1.5E-03
1.4E-03

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28
7.9 1.0E-02
60 1.3E-03
29
6.8 1.0E-02
60 1.1E-03
30
8.1 1.0E-02
60 1.4E-03
Average
9.4
NA
NA 1.6E-03
Table 4 on the previous page shows the data collected during the
inorganic trials. It also shows the averages for the moles of vitamin C and the
difference of iodine. It says NA, or not applicable, for the averages of the molar
solution of iodine and the volume of juice because they are constants.
Figure 5. Organic Juice After Titration

Figure 5 is a picture of the organic juice after the titration. Before the
titration the organic juice had an orange color, and after it had a greenish,
blackish color.
Figure 6. Inorganic Juice After Titration
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Figure 6 is a picture of the inorganic juice after the titration. Before the
titration the inorganic juice had a light yellow color, and after the titration it had a
darker yellow color with a blackish tint.
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Data Analysis and Interpretation

Data was collected by doing a titration on freshly squeezed organic
orange juice, and store bought Minute Maid orange juice. A molarity calculation
was then performed to find the concentration of vitamin C was in the juice.
Exactly thirty trials were done on each sample, so by the central limit theorem the
data should be normal. However, after taking a look at the histogram, boxplot,
and normal probability plot of the data, it was decided that the data was not
normal. Even though the data was found to be abnormal, the test was performed
anyways due to the central limit theorem and the fact that the other requirements
for the test were met. The other requirements are that two simple random
samples were performed on the different juices, and the samples were from
independent populations. Since it is indeed not normal though, the test results
may not be completely accurate, and therefore not completely reliable. Because
of this a descriptive analysis was done as well.
One of the requirements for a two sample t- test was that the data had to
be normally distributed, or relatively large. The figures below display the graphs
and plots that were used to determine if the data was normal or not. A normal
probability plot for each set of data, a boxplot of the each data set, and a dot plot
for each data set were all used to determine the normality of the distribution of
the data.
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Figure 7. Organic Orange Juice Probability Plot

Figure 7 shows the normal probability plot of the organic orange juice
concentrations. The dots on the plot are relatively far away from the line showing
that the data is not normal.
Figure 8. Inorganic Orange Juice Probability Plot

Figure 8 shows the probability plot for the inorganic orange juice
concentrations. The dots are closer to the line than the ones shown in figure 7
showing that the inorganic juice data is more normal than the organic juice. The
dots are still relatively far from the line though, so the data is still not normal.
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Figure 9. Box Plots

Figure 9 shows the boxplots for the organic and inorganic juice
concentrations. In agreement with the probability plots in figure 4 and 5, neither
of the data sets are normal. This is shown by the disproportionate size of the
quadrants in both of the plots. Also in agreement with figures 7 and 8 the data for
the inorganic juice - shown as the boxplot on the top - is more normal than the
organic juice - shown as the boxplot on the bottom. The data for the organic juice
is skewed to the left showing that there are less data points in the left quadrants.
The data for the inorganic juice is skewed to the right, showing that there are less
data points in the right quadrants. It is seen that the boxplots barely overlap, so it
can be concluded that the data for the inorganic and the organic do not contain
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the same amount of vitamin C. Since the data for the inorganic juice is skewed to
the right and the data for the organic juice is left skewed it can also be concluded
that the organic orange juice contains more vitamin C than the inorganic orange
juice.
Figure 10. Organic Orange Juice Dot Plot

Figure 10 shows the dot plot for the data collected during the organic
orange juice trials. The plot shows that the data points are mostly in one area to
the left, but then dies out and way over to the right there are more data points.
This shows that the data is skewed to the right, meaning that there are less data
points in the right area of the plot, which agrees with figure 9.
Figure 11. Inorganic Orange Juice Dot Plot

Figure 11 shows the dot plot for the data collected during the inorganic
juice trials. The data points are relatively spread out, but there are slightly more
in the left area of the plot. Because of this the data is skewed to the right
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meaning that there are less data points in the right area of the graph, which
agrees with figure 9.
A Two Sample T-Test was performed on the data. This test is used to
determine if two population means are the same (Two Sample T-Test for Equal
Means). The two populations in this experiment are the concentrations of
vitamin C in the organic and inorganic juices. The test was used to determine if
the concentration of vitamin C in the organic juice was significantly greater than
the concentration of vitamin C in the inorganic juice.
The test uses two hypotheses to talk about the results of the test. The first
is the null hypothesis, which is the same for every Two Sample T-Test. The
second is the alternative hypothesis. The alternative hypothesis is the hypothesis
the researchers, or people performing the test, believe the results are going to
show.
H 0 : 1=2
Figure . 12. Null Hypothesis
Figure 12 shows the null hypothesis.
hypothesis.
H 0 shows that it is the null
1 shows the mean of the first variable which is the mean of the
organic juice concentration.
2 shows the mean of the second variable which is
the mean of the inorganic juice concentration. The null hypothesis states that the
mean values of vitamin C concentration in the inorganic and organic juices are
equal.
H a : 1> 2
Figure 13. Alternate hypothesis
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Figure 13 shows the alternate hypothesis. As in figure 4, This alternate

hypothesis shows that
H a shows that it is the alternative hypothesis,
shows the mean of the the organic juice concentration, and
2 shows the mean
of the inorganic juice concentration. This alternate hypothesis shows that the
concentration of the vitamin C in the organic juice concentration is greater than
the concentration of the vitamin C in inorganic juice concentration.
Figure 14. Calculator T-test.

Figure 14 displays the results after the 2 sample t-test was done on the Tinspire. The t-value found was 8.0, and the p-value found was 2.6* 10 -4. Since the
p - value is lower than the alpha level of 5%, 0.05, the null hypothesis was
rejected. This means that the difference between the concentration of vitamin C
in the organic orange juice and concentration of vitamin C in the inorganic orange
juice was statistically significant. Although this supports the hypothesis that the
organic orange juice had a higher concentration of vitamin C than the inorganic
29

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21 May 2015
orange juice the test still may not be very reliable. Since the test was not
completely reliable a descriptive analysis was also done.
The mean of the vitamin C concentration in the organic juice is
with the standard deviation of
4 .1 104
concentration in the inorganic juice is
1.6 103
, and the mean of the vitamin C

4
9.1 10
with the standard deviation of
2.0 104 . On average, the organic orange juice yielded more vitamin C than
the inorganic juice. Because of this, it can be concluded that there is more
vitamin C in freshly squeezed organic orange juice then there is in store bought
Minute Maid orange juice.
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21 May 2015
Conclusion
The objective of this experiment was to find out whether processed
inorganic Minute Maid orange juice had differing amounts of vitamin C - or citric
acid - than freshly squeezed organic orange juice. Using descriptive statistics,
box plots, normal probability plots, and dot plots, the normality of the data points
as well as any trends or patterns in the data were examined. Then to analyze the
concentration of vitamin C in the two juices, a statistical two sample t- test was
used to compare the average molarity of the vitamin C in the juices. The
hypothesis, that the freshly squeezed organic orange juice would contain more
vitamin C than the processed inorganic orange juice, was accepted. The organic
orange juice did contain more vitamin C concentration than inorganic orange
juice.
The data along with the results of the two sample t-test supported the
hypothesis. An analysis of the averages of the calculated molarity for the trials,
the organic orange juice had a higher molarity with 1.6 x 10 -3 moles per liter of
vitamin C compared to the inorganic average of 9.1 x 10 -4 moles per liter of
vitamin C. The two sample t-test also supported the hypothesis. When the test
was executed the p-value was found to be 2.6 x 10 -10 power, approximately zero,
31

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which is significant on both the 5% alpha level. The p-value of the two sample ttest suggested that the averages of the inorganic and organic orange juices were
not the same, which rejects the null hypothesis that suggests that the average
vitamin C concentrations were the same, and accepting the alternative
hypothesis that suggests that the average concentration of the vitamin C found in
the organic orange juice is higher than the average concentration of the vitamin
C found in the inorganic orange juice.
When the data points were graphed on the box plots,dot plots , and the
normal probability plots the data was not normally distributed. Since there were
30 data points, the probability plots should have been normal. Since it was not
normal, it showed that while the experiment was being conducted some errors
were made. The data of the organic orange juice appeared skewed to the left
while the data of the inorganic orange juice appeared to be skewed to the right.
This concluded that the results of the two sample t-test were not conclusive
enough to draw a significantly reliable answer. Although the data points were not
normal there were thirty data points for both the organic and inorganic orange
juices, which should be have a normal distribution as stated in the Central Limit
Theorem, the test was still conducted to see if it would disprove the hypothesis.
Along with the statistical test, descriptive statistics were also used to analyze the
data. As seen in Figure 3 of the data analysis portion of the paper, it was
apparent that there is a significant difference in the concentrations of the vitamin
C. Because the box plots barely overlap, it can be noticed that the average
32

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21 May 2015
concentrations are varying because the medians of the data are not the same.
According to the placement of the box plots, it can be determined that the freshly
squeezed organic orange juice contained more vitamin C than the store bought
inorganic orange juice because the organic orange juice is right skewed on the
box plot. The inorganic orange juice is left skewed suggesting that the inorganic
orange juice contained less vitamin C.
The results of this experiment can be validated with other recent research.
Recent research in California found that fresh squeezed orange juice had 40%
more vitamin C than premium orange juice and 40% more vitamin C than orange
juice made from frozen concentrate (Harrison). Another experiment had found
that fresh squeezed orange juice had more vitamin C than other processed juices
tested (Terpstra) . Fresh squeezed orange juice is more likely to contain a higher
amount of vitamin C because the juice is not stored, preserved, or exposed to
oxygen, speeding up the reduction of nutrients in the juice. However, there is still
a chance that store bought orange juice could contain a higher amount of vitamin
C because producers may add vitamin C concentrate to the juice while it is being
stored.
Since the data was not normal, it can be inferred that the experiment was
effected by unknown variables. One error that could have affected the data was
the molarity of the second batch of iodine. The second batch of iodine, that was
made on 21st of the month, was about half the molarity that it should have been.
This made the amount of iodine needed per trial much larger than needed in
33

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21 May 2015
trials previously conducted. To account for the error, the formula used to find the
molarity of each solution was modified to the new molarity for the trials affected
by the new iodine. If this error would have gone unnoticed the experiment would
have been null and void. Another error that was made was the making of the
organic orange juice. The organic orange juice for some trials was not made on
the day the trial was conducted. The vitamin C in orange juice decreases over
time, so the concentration of vitamin C of orange juice that was not made on the
day of the trial could have been affected.
Along with the several errors made, a few problems also occurred while
the experiment was taking place. One problem that could not have been avoided
was mold found on the oranges. Towards the end of trials mold was found on
several of the oranges being squeezed for the organic juice. The oranges found
with mold on them were not used in the experiment, but they still could have
affected the vitamin C in the oranges that were used. Although the mold was not
seen until the last few days of trials, it could have been on the oranges used
previously but went unnoticed. Another problem that was encountered was with
the time allotted for trials.
The design of the experiment had many benefits as well as drawbacks. A
benefit of the design was how easy it was to follow. Two titrations could be done
at the same time while the solutions for the next titrations were being created. By
carrying out the experiment this way, a multitude of trials were able to be done
within the time allotted. A drawback of the experimental design was the time in
34

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21 May 2015
between making new batches of iodine. The iodine was not made fresh every
day, so it had time to deteriorate which could have affected the data. Also the
burette counts down from fifty, so instead of the iodine starting at the 10 mL line,
it started at the 40 mL line. That made figuring out how much iodine used during
the titration slightly confusing.
Overall the design of this experiment made the experiment easier to
execute. One change that could be made to this experiment is the type of juice
being used. Although this experiment is specific to orange juice, juices from other
fruit such as lemons and grapefruits could be used. Another change that could be
made to this experiment is using different types of oranges and different brands
of orange juices. This experiment could be very beneficial to orange juice
consumers as well as orange producers. Orange producers could present this
information to sell more oranges, and this may persuade orange juice consumers
to make their own orange juice instead of buying store brand orange juice.
35

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21 May 2015
Appendix A Sample Calculations

Finding the molarity
H C 2 H 3 O2 + NaOH NaC 2 H 3 O2+ H 2 O
If 5.0 mL of 0.50 M NaOH is used to neutralize 10.0 mL of HC 2H3O2, what is the
molarity of the acid solution?
M a V a=M b V b
M a 10=0.50 5
M a=
0.50 5
10
M a=.25
Figure 1. Sample calculation of finding molarity

Figure 1 displays a sample calculation to finding the molarity of a solution.
is the molarity of the first solution.
the molarity of the solute and the
V a is the volume of the first solution.
Ma
M b is
V b is the volume of solute. In this example
the molarity of the solution is being found and the both volumes and the molarity
of the solute are all given.
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21 May 2015
Two Sample T-Test

t=
t=
t=
x 1x 2
s 21 s 22
+
n1 n 2
105
2 1.5
+
50 50
5
3.5
50
t=18.8982
Figure 2. Sample 2 sample t-test
Figure 2 displays a sample calculation of a 2 sample t-test.
the first set of data, which is the organic juice, and
second set of data, which is the inorganic juice.
the first set of data and
x 1 is the mean of
x 2 is the mean of the
s 1 is the standard deviation of
s 2 is the standard deviation of the second set of data.
n1 is the number of data points in the first set of data and
n2 is the number of
data points in the second set of data points. In the sample calculation the t value
is 18.8982.
t=
t=
x1 x2
s 21 s22
+
n1 n2
( 1.6 103 ) (9.1 104 )
4.1270 104 1.9638104

+
30
30
37

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21 May 2015
Figure 3. t-test
Figure 3 shows the formula and substitution for the test performed. The
formula is the same as the one used in figure 2 on the previous page, for help go
to figure 2. In this test the t-value is
2.619711010 . In other circumstances,
after the t value is found, the degrees of freedom, DF, would need to be found to
carry on. The degrees of freedom is 1-n, n being the number of data points. After
finding the degrees of freedom, the p-value of would be found using Table C. but
in the situation the p value cannot be found because the t value is too small to be
found on table C.
Appendix B Randomization
Materials:
Ti-nspire
Procedures:
1.
Turn on Ti-nspire and open a new calculator page
2.
Press the menu button and scroll to choice 5: Probability and press enter
3.
Scroll to choice 4: Randomization and press enter
4.
Scroll to choice 2: Integer and press enter
5.
The calculator page will say randInt(). In the parenthesis enter one
comma two comma one meaning that the randomization will start
at one
and end at two and one number will be shown when enter is hit. If
the
number one is shown the inorganic orange juice will be titrated
first, if the
number two is shown the organic orange juice will be
titrated first
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21 May 2015
6.
Repeat steps 1-5 for the number of trials being done
Appendix C Preparing the Hydrochloric Acid

Materials:
Beaker (25mL)
Graduated cylinder (10mL)
Distilled water
Hydrochloric Acid, HCL
Glass container with a lid
Procedure:
1.
Obtain Materials and put on goggles and rubber gloves to ensure safety
2.
Using the graduated cylinder measure out 7.5 mL of water and pour it into
the beaker
3.
Also using the graduated cylinder measure out 2.5 mL of hydrochloric

acid, HCL, and pour it in the beaker
39

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21 May 2015
4.
Swirl the beaker to mix the hydrochloric acid and the water
5.
After the solution is mixed, pour the solution into the glass container and
securely place the lid on top
6.
Repeat process when hydrochloric acid is low
7.
To dispose of hydrochloric acid pour down the drain. It is safe to do so

because the molarity is very low
Appendix D Preparing the Iodine

Materials:
Distilled water
Iodine, I2
Dark glass bottle with a lid
Procedure:
1.
Obtain Materials and put on goggles and rubber gloves to ensure safety
2.
Using the graduated cylinder measure out 240 mL of water and pour it into
the bottle
3.
Also using the graduated cylinder measure out 60 mL of hydrochloric acid,

HCL, and pour it in the bottle
4.
Secure the lid on the bottle and shake it to mix the iodine with the water
40

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21 May 2015
5.
Repeat process when iodine is low
6.
To dispose of iodine pour down the drain with large amounts of water. It is
safe to do so because the molarity is very low
Appendix E Preparing The Starch Solution

Materials:
Distilled water
Soluble Starch (2 grams)
Glass bottle with a lid
Scale
Weigh Boat
Scoopula
Procedure:
1.
Using the scoopula and the weigh boat, on the scale measure out 2 grams
of soluble starch **Note** make sure to reset the scale after
41

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21 May 2015
placing the
weigh boat on top of it so the weight of the weigh boat is not
counted
2.
Using the graduated cylinder measure out 100mL of water and pour it into
the bottle
3.
Next pour the soluble starch into the bottle containing the water
4.
Secure the lip on the bottle and swirl the solution to mix the starch and
water
5.
Repeat procedure if starch solution runs low
6.
To dispose of the starch solution, pour down the drain
Works Cited
"1.3.5.3. Two-Sample T-Test for Equal Means." 1.3.5.3. Two-Sample T-Test for
Equal Means. N.p., n.d. Web. 03 May 2015.
<http://www.itl.nist.gov/div898/handbook/eda/section3/eda353.htm>.
"Acid-Base Titrations: Chapter 24." Acid-Base Titrations: Chapter 24. N.p., n.d.
Web. 22
Mar.2015.i<http://dwb.unl.edu/Teacher/NSF/C12/C12Links/users.ev1.net/
7Evklawinski/chpt24ntitration.html>.
42

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21 May 2015
"Arrhenius Acid-Base Direct Titration Calculations." Chemistry Tutorial : Arrhenius

Acid-base Direct Titration Calculations. N.p., n.d. Web. 18 Mar.
2015 <http://www.ausetute.com.au/titrcalc.html>.
A Titration Is A Means Of Quantitative Analysis In Which The Substance To Be,
Measured (In A Liquid Solution) Is Reacted Stoichiometrically With
Another Reagent, and (Called A Titrant) Until It Has Completely Reacted.
The End Of The Reaction Is Usually. H C C O(n.d.): n. pag. Web.
<http://chemlab.truman.edu/CHEM100Labs/THE%20DETERMINATION
%20OF%20CITRIC%20ACID.pdf>
"ChemLab - Techniques - Titration." ChemLab - Techniques - Titration. N.p., n.d.
Web.18 Feb.2015.
<http://www.dartmouth.edu/~chemlab/techniques/titration.html>.
Chan, Casey. "Dirty Little Secret: Orange Juice Is Artificially Flavored to Taste
Like Oranges." Gizmodo. Gizmodo, 29 July 2011. Web. <
http://gizmodo.com/5825909/orange-juice-is-artificially-flavored-to-tastelike-oranges>
Crosta, Peter. "What Is Scurvy?" Medical News Today. MediLexicon
International, 26 Sept. 2014. Web.
<http://www.medicalnewstoday.com/articles/155758.php>
Dishaw, Thomas. "A Few Reason To Dump That Carton Of Orange Juice Down
The
Drain |."Govtslaves. N.p., n.d. Web. 22 Mar. 2015.
<http://govtslaves.info/reason-dump-carton-orange-juice-drain/>.
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G., and 24 April 2005 Christina Terpstra: Vitamin C In Orange Juice, Page 1 Of.
VITAMIN C IN ORANGE JUICE Christina Terpstra PURPOSE (n.d.): n.
pag. Web.
Hamilton, Alissa. Squeezed: What You Don't Know about Orange Juice. New
Haven: Yale UP, 2009. Print.
Hawley, Gessner Goodrich, and Richard J. Lewis. Hawley's Condensed
Chemical Dictionary. 12th ed. New York: Van Nostrand Reinhold, 1993.
1275 Pages. Print.
Lindsay. "Oranges to Oranges: Which Orange Juice Has the Most Vitamin C:
Fresh Squeezed, Premium, or Frozen Concentrate?" Abstract (2008): n.
pag. Print.
Medical News Today. MediLexicon International, n.d. Web. 20 May 2015.
<http://www.medicalnewstoday.com/articles/155758.php>.
"Pasteurization." Pasteurization. International Dairy Foods Association, n.d. Web.
"Physical and Chemical Properties." Encyclopedia Britannica Online.
Encyclopedia Britannica, n.d. Web. 19 Mar. 2015.
<http://www.britannica.com/EBchecked/topic/292625/iodineI/280159/Physical-and-chemical-properties>.
Science, College Of. Determination of Vitamin C Concentration by Titration(n.d.):
n.pag.Webi<http://www.outreach.canterbury.ac.nz/chemistry/documents/v
itaminc_iodine.pdf>
"Scurvy - Symptoms ." Scurvy. N.p., n.d. Web. 20 May 2015.
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"Vitamin C In Fruit Skins." The British Medical Journal 2.4160 (1940): 424. Web.
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Eskuri Rupert Tablada

Eskuri Rupert Tablada

Eskuri Rupert Tablada

amount of vitamin C to reduce the possibility of vitamin C deficiency such as

Eskuri Rupert Tablada

The purpose of this experiment was to determine the vitamin C content in

Eskuri Rupert Tablada

Conducting a titration lab on vitamin C in orange juice was chosen to find

Figure 1. The structure of vitamin C

Eskuri Rupert Tablada

Eskuri Rupert Tablada

where the first C6 H 8 O6 is ascorbic acid,

the dehydroascorbic acid which turns the blue/black color.

Figure 2. Titration Curve

M a is the molarity of the acid,

M b is the molarity of the base, and

is the volume of the acid,

V b is the volume of the base ("Acid-

Eskuri Rupert Tablada

If 5.0 mL of 0.50 M NaOH is used to neutralize 10.0 mL of HC 2H3O2, what is the

Eskuri Rupert Tablada

Eskuri Rupert Tablada

Eskuri Rupert Tablada

Eskuri Rupert Tablada

Eskuri Rupert Tablada

Data and Observations

Eskuri Rupert Tablada

Eskuri Rupert Tablada

Inorganic Orange Juice Observations/Comments

Eskuri Rupert Tablada

Organic Orange Juice Observations/Comments

Eskuri Rupert Tablada

Eskuri Rupert Tablada

Eskuri Rupert Tablada

Organic Orange Juice Observations/Comments

Table 3. Inorganic Orange Juice Data

Eskuri Rupert Tablada

Inorganic Orange Juice

Eskuri Rupert Tablada

Organic Orange Juice

Eskuri Rupert Tablada

Figure 5. Organic Juice After Titration

Figure 6. Inorganic Juice After Titration

Eskuri Rupert Tablada

Eskuri Rupert Tablada

Data Analysis and Interpretation

Eskuri Rupert Tablada

Figure 7. Organic Orange Juice Probability Plot

Figure 8. Inorganic Orange Juice Probability Plot

Eskuri Rupert Tablada

Figure 9. Box Plots

Eskuri Rupert Tablada

Figure 10. Organic Orange Juice Dot Plot

Figure 11. Inorganic Orange Juice Dot Plot

Eskuri Rupert Tablada

H 0 shows that it is the null

organic juice concentration.

2 shows the mean of the second variable which is

Eskuri Rupert Tablada

Figure 13 shows the alternate hypothesis. As in figure 4, This alternate

H a shows that it is the alternative hypothesis,

shows the mean of the the organic juice concentration, and