Analysis of Ascorbic Acid in Fruit by Iodometric Titration

Abstract

The analysis of ascorbic acid was done in two media, whole fruit and orange juice. The
concentration of the iodide as provided by California State University, Fullerton was found to be 0.0107
(±0.001) M iodine iodine using Iodometric titration techniques with known grams of ascorbic acid. The
known iodine concentration was then used to determine the concentration of ascorbic acid in Tropicana
Pure Premium 100% Pure Orange Juice with calcium and vitamin D and a whole orange. The Iodometric
titration, performed with a large buret, was then compared to other large burets and smaller burets to to
test the precision and accuracy. The large buret yielded 109.31 (±3) mg of ascorbic acid per 8 ounces of
Tropicana Pure Premium Orange Juice while a whole fruit contained be 42.53 (±0.8) milligrams of
ascorbic acid per 1 fruit. Subsequent F and T test show that the standard deviation of each measurement
was statistically different while the T test show that the means are not statistically different. This suggests
that, though the burets differ in precision, but they both arrive at the same true value. It is hypothesized
that the smaller burets can replace the already accurate larger burets and improve upon the precision and
accuracy. It was found that the smaller burets cannot replace the larger burets because the precision and
the accuracy was deemed statistically different compared to the accurate large buret.

Introduction

Humans, apes, and guinea pigs are the only organisms that require vitamin c to be consumed
through the diet because a loss of important enzymes for vitamin c biosynthesis. Vitamin C, though, can
be found in a variety of fruits and vegetables such as oranges, kale, and watermelon and is abundant in
the normal American diet. Ascorbic acid has important function in the human body improving iron
absorption and resistance to infection. Though there is no evidence to support the resistance to infection
claim made popular by Linus Pauling, there is unprecedented evidence on ascorbic acids role to form
1
collagen, absorb iron through the diet, and serve as an antioxidant . Since there is such an importance
for having vitamin C in the body in the form of ascorbic acid, prevalent disease comes about when a
1
deficiency occurs or a mutation for the activation of vitamin C . Scurvy is a very uncommon disease in
the United States but still affects 20,000 US citizens per year characterized by muscle pain, joint pain,
2
fatigue, red dots on the skin, and bleeding of gums . Muscle pain and fatigue are direct symptoms of
what may be associated with an iron deficiency from poor absorption of iron via a ascorbic acid
deficiency. Vitamin C has a main role providing a foundation for the adsorption of dietary iron. In
addition, ascorbic acid acts as an antioxidant, by removing superoxides in the body made from the
electron transport chain where the electrons individually are donated to the terminal electron acceptor,
1
O2 .

Figure 1: The molecular formula for ascorbic acid is C6H8O6. Ascorbic acid gets reduced to
3
dihydroascorbic acid when reacted with iodine ion .

The National Institutes of Health determine that the recommended daily amounts of vitamin C vary
with age where an average male should take in 90 mg and an average woman should intake 75mg.
Those who are diagnosed with scurvy are likely to get below 10 mg of vitamin C for consecutive weeks
and can be fatal if left untreated. Supplementing with ascorbic acid in pill form or receiving it from the diet
is the best treatment option for those with scurvy but can be harmful if not closely monitored. Many
supplements that contain vitamin C have around 1000mg per serving. Using both supplements and
receiving vitamin C from the diet may cause harmful effects such as diarrhea, nausea, and stomach
 4
cramps . Other more fatal symptoms of vitamin C overdose is hemochromatosis which his characterized
by the damage of body tissues through the continuous overload of absorption of iron in the cells.
An average orange of 131g will have approximately 70 mg of vitamin C while the average lemon
5
will have around 50 mg . The determination and analysis of ascorbic acid in the citrus fruits will be found
by using iodometric titration. Iodometric titration are similar to normal titration but the titrant involves an
iodine solution and a starch indicator. When iodine reacts with the starch indicator, a deep dark violet
color forms due to the complex iodine makes with starch. The iodine titrant will be in the form of
potassium iodate, KIO3 in the presence of a strong acid such as KI. The reaction that occurs between
KIO3 and KI, a diatomic iodine molecule, I2 is released into the system. This can be seen in equation 1
where 3 I2 are freed from the potassium iodide.
+ +
KIO3 + 5KI + 6H → 3I2 + 6K + 3H2O (1)

Much like acid base titrations, these are done in aqueous conditions so water is also to be
expected product and will not affect the results of the iodometric titration. As with all titrations, end point
detection is usually difficult and should be done before analyzing the analyte which in this case is
ascorbic acid. As stated before, iodine will react with starch to form a blue black complex that is easy to
detect. Iodine, though, can be used as a natural indicator because it forms a brown color but the iodine
starch complex forms intense blue colors as seen in equation 2.
-
2I + starch à blue-black complex (2)

As stated, the purpose of a titration is to find the amount of moles that are in the initial analyte
solution. Ascorbic acid will need to be selectively analyzed while all other molecules such as sugar and
fiber must not be. Iodometric titrations are an ideal method to analyze ascorbic acid because ascorbic
acid reacts with I2 that is formed from the original reaction in equation 1 with potassium iodide. Ascorbic
acid, C6H8O6, as seen in figure 1, can be be oxidized to dehydroascorbic acid, C6H6O6, through this
-
reaction and the product with be I . Equation 3 details the oxidation of ascorbic acid to dehydroascorbic
acid
- +
C6H8O6 + I2 → C6H6O6 + 2I + 2H (3)

Once the iodine is made, it can react with the starch to form the blue black complex. This
complex is easily detected because a great part of the titration will be done in orange juice which is an
orange color. Without the starch iodine complex, the orange color of the orange juice may mask the end
point. The starch iodine complex will show a dark brown color when the blue-black complex mixes with
the natural color of orange juice. The selectivity comes from the ascorbic acid’s ability to be reduced by
iodine and iodine picks up the electrons. Only the ascorbic acid will react with the iodine while everything
else will be left constant. The iodine then reacts with the starch that is present in the mixture and forms a
blue black complex.

Using the concepts of titrations, the initial moles of ascorbic acid can be used based on the
volume of the titrant used. In essence, the moles of titrant is equal to the moles of analyte in solution at
the point in time where the indicator changes permanently to one color. In this case, when the starch
binds to the iodine making the blue black complex. For this experiment, orange juice and whole orange
fruit will be compared on their contents of ascorbic acid in milligrams.

Experimental

The procedures of the lab manual provided by California State University, Fullerton were followed
5
precisely to determine the concentration of ascorbic acid in fruit juice and whole fruit . A 200 mL stock
iodine solution was obtained from the lab and was labeled with name and date. Only one bottle of iodine
solution was given and there were no extra bottles. To analyze ascorbic acid using the ascorbic acid
iodine system, the concentration of iodine must be determined. The iodine solution was made in one
large batch and split up into 16 students in the lab, therefore, the iodine solutions should be the same.
However, it is important to determine one’s own iodine solution to ensure accuracy of the experiment.
This is easily done by completing a titration with the iodine of unknown concentration with known amounts
of ascorbic acid.

0.100 g of 100% ascorbic acid was obtained from the lab and used for the Iodometric titration to
determine the concentration of iodine in the iodine solution. With the addition of 20 drops of starch, the
analyte containing the ascorbic acid and the starch were ready for the titration. 25.00 mL of each analyte,
whether containing ascorbic acid, orange juice, or whole fruit, was measured using a 25.00 mL volumetric
pipet. The found iodine concentration can now be used to determine the concentration of ascorbic acid in
store bought Tropicana Pure Premium 100% Pure Orange Juice with calcium and vitamin D purchased
th th
on November 18 2015 with a December 24 2015 expiration date. The nutrition facts label states a
single 8 fluid ounce serving will contain 120% of the daily value of vitamin C, ascorbic acid. Titrations
were performed until 3 titrations had a ratio of standard deviation (RSD) that was below 10 parts per
thousands. Reading the meniscus with a buret reader, made of an index card and a black line, ensured
proper and repeatable readings. All data was recorded in excel.

The analysis of whole fruit was then done to determine the ascorbic acid in whole fruit. Using the
same procedures and completing 3 titrations with an RSD that is below 10 parts per thousand, the
comparison whole orange to store bought orange juice was allowed. The 6 whole oranges were
purchased at Ralph’s Grocery Store and juiced the same day using the juicer provided in lab.

The buret used in all titrations was considered a large buret with half the class having a large
buret and the other half having a small buret. The validity of data collected from the buret was
determined through statistical analysis using the F test and the T test. Secondly, it was a necessity to
determine the validity of data collected from the Iodometric titrations comparing it to the other large
burets, excluding the small burets. Third, statistical analysis was performed to compare the iodine
concentrations and ascorbic acid concentrations from orange juice of the large and small burets. The F
test and T test were done to compare the standard deviations and the means of each data set.

Results

Iodine Concentration

The iodine concentration was determined to ensure the correct concentration was used for the
Iodometric titration. To calculate the iodine concentration, known amounts of pure ascorbic acid was
used to titrate with. Using the 0.100g of ascorbic acid and 20 drops of starch as the analyte, the
Iodometric titration was done with 3 successive trials with and RSD value less than 10 ppt. The volumes
of iodine were found to be 0.00552 L, 0.00554 L, and 0.0055 L to reach the end point of the colorless
ascorbic acid solution. The end point was determined to be a pale blue color as compared to the study
6
done by University of Canterbury .

These volumes would be used to determine the concentration of iodine solution using equation 4
where M1 will stand for the molarity of acid and V1 will be the volume of ascorbic acid added in the
analyte. The M2 and V2 will represent the iodine concentration and volume used, respectively.

M" V" = M% V% (4)

M&'() V&'() = M(*)(+, V(*)(+,
1 mole ascorbic acid 1
0.100g ascorbic acid × (0.02500L)
170.12 g ascorbic acid 0.25L
M(*)(+, =
0.00552 L iodine

The Macid was found by using the mass of ascorbic acid used for the analyte, 0.100g, and multiplying it by
the molar mass to achieve a value for the number of moles used. The moles of ascorbic acid can then be
multiplied by the volume of ascorbic acid used to make the solution, 250 mL, taking into account the units
and converting it to L. Now, 25 mL, or 0.02500L, were used as the analyte for each Iodometric titration
which would serve as the volume of ascorbic acid used. The volume of iodine found for the first trial was
0.00552L which would be used in equation 1 to solve for the molarity of iodine. The molarity of iodine for
each trial was found to be 0.0106 M, 0.0106 M, and 0.0107M iodine.

The molarity of iodine of each trial was used to determine the RSD in parts per thousand and to
5
determine if the trials needed to be repeated .
HI
RSD = × 1000 (5)
J

The RSD is the ratio of standard deviation to the data set mean and multiplying by 1000. The average, x̄,
was found to be 0.01065 using the excel function, =AVERAGE. The standard deviation was calculated
7
using equation 6 .

JKJ L
sJ = (6)
+K"

0.0106 − 0.01064901 % + 0.0106 − 0.01064901 % + 0.0107 − 0.01064901 %

sJ =
3−1
sJ = 3.8584 × 10KT 𝑀 𝑖𝑜𝑑𝑖𝑛𝑒
-5
The standard deviation was found to be 3.8584 x 10 M iodine. Once the standard deviation and the
mean were found, the RSD for the iodine concentration was found to be 3.62 parts per thousand which
was under the 10 parts per thousand standard. The 95% confidence interval was needed to determine
the validity of the iodine concentration. To calculate the confidence interval, equation 7 was used where t
is the student’s t and s is the standard deviation.

Confidence Interval = ts = 0.2776 3.8584 × 10KT = 0.000107 (7)

The student’s t was found to be 0.2776 using 95% confidence and 4 degrees of freedom where
7
degrees of freedom is the number of data points, n, subtract 2 . The confidence interval was determined
to be 0.000107 and, as a result, the iodine concentration was found to be 0.0107 (±0.001) M iodine at
95% confidence.

Orange Juice Concentration

Orange juice concentration was determined using the iodine concentration calculated before.
th th
The orange juice was purchased November 18 with an expiration date of December 25 the following
month. The orange juice concentrations were determined using the same procedures as the iodine
concentration. 3 trials were performed with the orange juice when 20 drops of starch was added to the
analyte. The 3 Iodometric titrations took 6.21 mL, 6.10 mL, and 6.15 mL of 0.0107 (±0.001) M iodine to
reach the end point. The end point was compared to the same paper where the color was a near sick
green, brown color.

Using these volumes of 0.0107 (±0.001) M iodine and equation 8, the concentration of ascorbic
acid in orange juice can be found. Unlike the previous equation where Miodine was calculated, Macid was
needed to be determined. To calculate this, Miodine was found to be 0.0107 (±0.001) M iodine from
previous calculations. The volume of iodine used to reach the endpoint was placed as Viodine. In addition,
the Vacid used in each titration was 0.02500 L of orange juice.

M" V" = M% V% (8)

M&'() V&'() = M(*)(+, V(*)(+,
M(*)(+, V(*)(+,
M&'() =
V&'()
0.010649 𝑚𝑜𝑙𝑒𝑠 𝑖𝑜𝑑𝑖𝑛𝑒
(0.00621 𝐿 𝑖𝑜𝑑𝑖𝑛𝑒 )
𝑀`abc = 1 𝐿 𝑖𝑜𝑑𝑖𝑛𝑒
(0.02500 𝐿 𝑎𝑐𝑖𝑑)
 𝑀`abc = 0.00265 𝑀 𝑎𝑠𝑐𝑜𝑟𝑏𝑖𝑐 𝑎𝑐𝑖𝑑

Using equation 8 for all 3 trials, the ascorbic acid concentration was found to be 0.00265M, 0.00260M
and 0.00262 M ascorbic acid in the orange juice. The ascorbic acid concentrations of the orange juice
was then converted into a usable form for the RSD calculation, mg per 8 ounces. Using dimensional
analysis, the molarity of ascorbic acid was multiplied by the known conversion of 0.029573 liters in an
ounce seen in equation 9.

l.ll%m% nop `qaorsba `abc l.l%uTvw t "vm."% y `qaorsba `abc "lll ny z ""l ny `qaorsba `abc
× × × × = (9)
" t `qaorsba `abc " ox " nop `qaorsba `abc " y z z ox o{ or`|y} ~•ba}

This value was then multiplied by the molar mass of ascorbic acid and converted to milligrams.
The final answer was in milligrams per ounce and can be converted to milligrams per 8 ounces by
multiplying the top and bottom by 8. This value was the milligrams of ascorbic acid that was found per 8
ounces of orange juice. For the first trial, it was found to be 110 mg of ascorbic acid per 8 ounces of
orange juice. All trials performed underwent the same procedure using equation 8 and was found to be
110.42 mg, 108.33 mg, and 109.17 mg per 8 ounces of orange juice. These values can then be used to
calculate the RSD by using the average and the standard deviation of the 3 trials and multiplying by 1000
5
as shown in equation 10 .
qƒ
𝑅𝑆𝐷 = × 1000 (10)
„

The mean, x̄, was found to be 109.31 mg per 8 ounces of orange juice. The standard deviation, sx, was
found using equation 11 by taking the square root of the differences of sums of the mean and dividing by
7
the degrees of freedom .
„K„ L
𝑠„ = (11)
|K"

109.31 − 110.42 %
+ 109.31 − 108.34 % + 109.31 − 109.17 %
𝑠„ =
3−1
1.0489 𝑚𝑔 𝑎𝑠𝑐𝑜𝑟𝑏𝑖𝑐 𝑎𝑐𝑖𝑑
𝑠„ =
8 𝑜𝑢𝑛𝑐𝑒𝑠 𝑜𝑓 𝑓𝑟𝑢𝑖𝑡 𝑗𝑢𝑖𝑐𝑒

The standard deviation was found to be 1.05 milligrams of orange juice per 8 ounces of orange
juice. By using the mean and the standard deviation calculated from equation 11, the RSD can be
calculated using equation 10. The RSD was found to be 9.60 parts per thousand which is under the 10
parts per thousand standard.

The 95% confidence interval was performed by multiplying the student’s t and by the standard
5
deviation .

Confidence Interval = ts = 0.2776 1.05 = 3 (12)

The student’s t was the same student’s t found in the determination of iodine concentration where the
degrees of freedom is 6 minus 2, 4. For the 95% confidence interval, the student’s t was found to be
7
0.2776 . With the confidence interval, the orange juice was found to have 109.31 (±3) mg of ascorbic
acid per 8 ounces of fruit juice.

Whole Fruit Concentration

The concentration of ascorbic acid in a whole orange fruit was found using the same procedures
as the orange juice. The fresh bought orange from Ralph’s Grocery Store was purchased on the same
day and juiced using a juicer provided in lab. The Iodometric titration using the 0.0107 (±0.001) M iodine
solution was performed to determine the concentration of ascorbic acid using equation 12. Where Miodine
was calculated in the previous experiments to be 0.0107 (±0.001) M iodine and the volume of iodine used
in each titration was variable. Vacid was the amount of fresh squeezed orange juice used in the analyte.
The volume of analyte used was 25.00 mL measured by a volumetric pipet.

Š‹Œ•‹Ž• •‹Œ•‹Ž•
𝑀`abc = (13)
•‘’‹•
0.010649 𝑚𝑜𝑙𝑒𝑠 𝑖𝑜𝑑𝑖𝑛𝑒
(0.00666 𝐿 𝑖𝑜𝑑𝑖𝑛𝑒 )
𝑀`abc = 1 𝐿 𝑖𝑜𝑑𝑖𝑛𝑒
(0.02500 𝐿 𝑎𝑐𝑖𝑑)
𝑀`abc = 0.00284 𝑀 𝑎𝑠𝑐𝑜𝑟𝑏𝑖𝑐 𝑎𝑐𝑖𝑑

The concentration of the ascorbic acid in the whole orange fruit juice was found to be 0.00284 M ascorbic
acid. Performing equation 13 on the subsequent trials, the molarity of ascorbic acid was found to be
0.00284 M, 0.00286 M, and 0.00282 M ascorbic acid. These concentrations can then be used to
determine the milligrams of ascorbic acid in each fruit. Much like in the orange juice calculations,
dimensional analysis was used to determine the milligrams of ascorbic acid per fruit using the molarities
of ascorbic acid as shown in equation 14.

l.ll%z“ nop `qaorsba `abc l.%TTt ~•ba} "vm."% y `qaorsba `abc "lll ny “%.“v ny `qaorsba `abc
× × × = (14)
t `qaorsba `abc w {r•b” " nop `qaorsba `abc " y " {r•b”

The molarities of the ascorbic acid were multiplied by the total volume of juice taken from the 3
fruit that were juiced as seen in equation 14. This value was multiplied the molar mass of ascorbic acid
and converted to milligrams to achieve units of milligrams of ascorbic acid per 1 fruit. Each trial
performed was subject to equation 3 to convert each value into milligrams of ascorbic acid per 1 fruit. It
was found to be 42.47 mg, 42.85, and 42.28 milligrams of ascorbic acid per 1 fruit for the trials of
Iodometric titrations.

The RSD was needed to determine if additional trials were necessary to achieve a value that is
5
below the 10 parts per thousand standard . The RSD was determined through equation 15 where the
7
standard deviation and mean of the data set was needed .
qƒ
𝑅𝑆𝐷 = × 1000 (15)
„

The average of the 3 trials was found to be 42.53 milligrams of ascorbic acid per 1 fruit. The standard
deviation of the data set was determined using equation 16.

„K„ L
𝑠„ = (16)
|K"

42.47 − 42.53 % + 42.85 − 42.53 % + 42.28 − 42.53 %

𝑠„ =
3−1
𝑠„ = 0.2922

Using the mean, 42.53 milligrams of ascorbic acid per 1 fruit, and the standard deviation, 0.2922
milligrams of ascorbic acid per 1 fruit, the RSD can be calculated. The RSD was found to be 6.87 parts
per thousand which was below the 10 parts per thousand standard set in class.

The confidence interval at 95% was determined using equation 17 where the student’s t was
7
found using a degrees of freedom of 4 . This value was multiplied by the standard deviation found in
equation 16.

Confidence Interval = ts = 0.2776 0.2922 = 0.81 (17)

From this, the amount of ascorbic acid in a whole orange was found to be 42.53 (±0.8) milligrams of
ascorbic acid per 1 fruit.
Comparison of Data Collected to Class Data of All Burets

The data collected for iodine concentration and milligrams of ascorbic acid in 8 ounces of orange
juice were compared against the class data to determine if the data collected was an outlier. The class
data excludes my data to ensure the accuracy of the F and T test to be performed. All class data was
compiled into excel. To start, the average of the class data was found to be 0.0117 M iodine and 119.89
mg of ascorbic acid per 8 ounces of orange juice using function “=AVERAGE” in excel. The standard
deviation was needed to perform each test for comparison using the standard deviation function
“=STDEV” in excel. The standard deviation for the iodine concentration was found to be 0.00207 M iodine
and for the orange juice, 11.73 mg of ascorbic acid per 8 ounces of fruit juice. The class data confidence
7
intervals were found using equation 17 where the student’s t was 2.021 . Using the student’s t and
multiplying it by the standard deviation for the iodine concentration and the milligrams of ascorbic acid in
8 ounces of juice, the confidence interval is determined to be 1.17 (±0.004) M Iodine and 119.89 (±10)
milligrams of ascorbic acid in 8 ounces of fruit juice.

These values can now be used to determine Fcalc which compares the standard deviations of each data
set, the data set collected from the results section and the class data set shown in equation 18.

q–L l.ll%"L
Fa`pa•p`”}c = = = 2875 (18)
qLL w.zm×"l—˜

-5
The standard deviation for the data collected in this experiment was 3.86 x 10 M iodine and, for the
class, was 0.0021. For the F test, the larger of the two standard deviations is represented by being in the
numerator, s1, while the smaller standard deviation is in the denominator as s2. As a result, the Fcalculated
was determined to be 2875. The Ftable value associated with 48 degrees of freedom was not found. The
7
Ftable value used was 40 degrees of freedom, 19.5 .

The tcalculated was performed using equation 19 where the means are compared.

„– K„L |– |L l.l"lvKl.l""v w “T
t a`pa•p`”}c = = = 0.891 (19)
q™ŒŒš•• |– ›|L l.ll%l% w›“T

The means for each data set for the concentration of iodine were found to be 0.0117 and for the data
collected in this experiment, 0.0107. These values would be placed for x̄1 and x̄2. For n1, there were
trials used for the determination of concentration of iodine in this experiment. For the class data,
represented as n2, will have 45 data points. The data points come from the 15 other students whom have
7
3 trials each. Spooled is calculated through equation 20 .

ž– L (+– K")›žL L (+L K") w.zm×"l—˜ (w)›l.ll%lvL (“T)

sœ**•,) = = = 0.00202 (20)
+– ›+L K% “m

Having obtained spooled, tcalculated can now be calculated. It was found that tcalculated was 0.891. The ttable
7
value associated with having 40 degrees of freedom was 2.021 .

Comparison of the milligrams of ascorbic acid in 8 ounces of fruit juice was found using the same
tests. Using the standard deviation from the class data for the milligrams of ascorbic acid per 8 ounces of
fruit juice, an F test can be done. The standard deviation from the class data was found to be 11.73 while
the standard deviation found in this paper was 1.0489. The Fcalculated value was found to be 125 and the
Ftable 19.5. The Ftable value was obtained by having 30 degrees of freedom for s1 and 2 degrees of
7
freedom for s2 .

Using the mean of ascorbic acid 119.89 from the class and 109.31 obtained through the results,
7
the tcalculated value was found to be 1.55 while the ttable was found to be 2.78 . All values for the
comparison of the data obtained from this paper and the data obtained from the class was placed into
table 1.

Table 1: Comparison of Iodine concentrations between the class data and experimentally derived data
was shown in row 1. F Test show that the standard deviations are significantly different but the T test
show the means are not statistically different. Comparison of the orange juice between the class data
and experimentally derived data was shown in row 2. The F test show the standard deviations are
statistically different and the T test show that the means are not statistically different.

Different? Fcalculated Ftable tcalculated ttable

Iodine (M) No 2875 19.5 0.891 2.021

Orange Juice No 125 19.5 1.59 2.021

(mg/8oz)

Comparison of Data Collected to Class Data Holding Buret Size Constant

Half the class had a large buret and the other half had a small buret. The data collected for the
iodine concentration, orange juice, and whole fruit were collected using a large buret that has a larger
diameter. The other half of the class had a smaller buret for their data. This comparison will compare the
buret used for this paper, a large buret, to the other 7 large burets used during the experiment. The
average was obtained through the same procedure as the last comparison which was determined to be
0.0107 M iodine for the large burets excluding data points that were made in this paper. The average for
the data found in this paper was 0.0107 M iodine as well. The standard deviation for the large burets,
excluding the data found in the paper, was found to be 0.00239 M iodine. The standard deviation, stated
-5
above, was 3.86 x 10 M iodine. Using these two standard deviations with the class data as s1 and the
7
data found in the paper as s2, the F test can be performed in equation 21 .

q–L l.ll%wuL
Fa`pa•p`”}c = = L = 3851 (21)
qLL w.zm×"l—˜

The Fcalculated value for the iodine concentration for all large burets was determined to be 3851.
The Ftable value was found to be 19.4 by having 20 degrees of freedom for s1 and 2 degrees of freedom
7
for s2 . The number of data points, n1, was found by multiplying the 7 students with large burets by the 3
trials performed. Data points for n2 was found to be 3 data points.

With this calculated, the T test can be performed but a spooled value is needed. As seen in equation 22,
the spooled value was determined using the data point values listed above.

ž– L (+– K")›žL L (+L K") w.zm×"l—˜ (w)›l.ll%wuL (%")

sœ**•,) = = = 0.0022 (22)
+– ›+L K% %%

With the spooled calculated, the tcalculated can be performed using the means of the iodine concentrations.
Since the means were the same number, the decimal places were extended out to achieve a tcalculated
value that is not zero.

„– K„L |– |L l.l"lmTKl."lv“z w %"

t a`pa•p`”}c = = = 0.070 (23)
q™ŒŒš•• |– ›|L l.ll%% w›%"
The tcalculated value was found to be 0.070 and, according to Harris, the ttable value associated with the 22
7
degrees of freedom is 2.086 .

The milligrams of ascorbic acid in 8 ounces of fruit juice was also compared to the class data for
large burets only. The average for the milligrams of ascorbic acid in 8 ounces of fruit juice for the 7 other
large burets was 113.00 milligrams of ascorbic acid per 8 ounces of fruit juice with a standard deviation of
3.79 milligrams of ascorbic acid per 8 ounces of fruit juice. The average obtained from this paper was
found to be 109.31 with a standard deviation of 1.05 milligrams of ascorbic acid per 8 ounces of fruit juice.

These standard deviations were subject to an F test as well using the same amount of data
points, 21 and 3, respectively for s1 and s2. The Fcalculated value was found to be 13 with the Ftable value for
7
the amount of data points to be 19.4 .

The means for the milligrams of ascorbic acid in 8 ounces of orange juice was subject to a T test.
The Tcalculated was performed in the same way as the T test performed in equation 23. The Tcalculated value
7
found to be 1.65. The Ttable value for 20 degrees of freedom was 2.086 . These values were placed into
table 2 for comparison.

Table 2: Comparison of the class data with large burets only, excluding data points made in this paper,
with data points made in the paper. The F test for iodine concentration showed the standard deviations
are statistically different from each other but the T test shows the means are not statistically different. For
the analysis of ascorbic acid in orange juice, the F test shows the standard deviations are not statistically
different and the T test shows the means are not statistically different.

Different? Fcalculated Ftable tcalculated ttable

Iodine (M) No 3581 19.0 0.070 2.086

Orange Juice No 13 19.4 1.65 2.021

(mg/8oz)

Comparison of Large Burets to Small Burets

The comparison of the large burets and the small burets was performed to identify the differences
of accuracy and precision between the burets. Recall, half the class of 16 students have the small burets
and the other half have large burets. It was hypothesized that the smaller burets would have higher
precision and accuracy than the larger burets. Examining the averages, the larger burets show an
average iodine concentration of 0.0107 M iodine with a standard deviation of 0.00223 M iodine. The
smaller burets show a larger iodine concentration of 0.0126 and a smaller standard deviation 0.00125 M
iodine.

An F test can be performed by comparing the standard deviations of the data taken from the large
burets and the small burets for the iodine concentration.

q–L l.ll%%wL
Fa`pa•p`”}c = = = 3.18 (24)
qLL l.ll"%TL

Using the larger of the 2 standard deviations as s1 and the smaller one as s2, the Fcalculated value shows to
be 3.18. With 48 data points, the degrees of freedom was found to be 22 for each data set, though, the
7
table only allowed up to 20 with a Ftable value of 2.12 .

A T test was needed to determine if the means were statistically different but a spooled calculation
was to be calculated first as shown in equation 25.
 ž– L (+– K")›žL L (+L K") l.ll%%wL (%“)›l.ll"%TL (%“)
sœ**•,) = = = 0.0018 (25)
+– ›+L K% “m

Once the spooled value was found, a T test can be performed with the means of the iodine concentration
from the large burets and the iodine concentrations from the small burets.

„– K„L |– |L l.l"lvwKl.l"%Tz %“ %“
t a`pa•p`”}c = = = 3.53 (26)
q™ŒŒš•• |– ›|L l.ll"z %“›%“

The tcalculated value was found to be 3.53 and 46 degrees of freedom, the ttable value was found to be 2.021
7
where only 40 degrees of freedom were used shown in equation 26 . These values were placed into
table 3 for comparison between the large burets and the small burets.

The milligrams of ascorbic acid in 8 ounces of orange juice was also analyzed for the comparison
of large burets with small burets. The average concentration found for the large burets was 112.54
milligrams of ascorbic acid in 8 ounces of orange juice with a standard deviation of 3.76. The smaller
buret yielded a larger value of 125.46 milligrams of ascorbic acid in 8 ounces of fruit juice with a
significantly higher standard deviation of 13.02.

The F test using the small burets as s1 and the larger burets for s2 to get a value larger than one
yielded an Fcalculated value of 12.00. Using 20 degrees of freedom for both data sets, the Ftable value is
7
shown to be 2.12 .

The T test was performed comparing the means of the large burets and the small burets. After
determining the spooled value of 9.75 using equation 25, the Tcalculated was calculated to be 4.68. Values
that were obtained through the comparison the large burets with the small burets were compiled into table
3.

Table 3: Comparison of large burets with small burets was shown using iodine concentration (M) and
milligrams of ascorbic acid in 8 ounces of orange juice. The F test shows that the the large burets and
the small burets have statistically different standard deviations when measuring iodine concentration.
Likewise, the means are statistically different as when measuring iodine concentrations. For the orange
juice, the large burets and the small burets differ both in standard deviation and mean.

Different? Fcalculated Ftable tcalculated ttable

Iodine (M) Yes 3.18 2.12 3.53 2.776

Orange Juice Yes 12.00 2.12 4.68 2.021

(mg/8oz)

Discussion

Iodine Concentration

The determination of iodine concentration was the first step in analyzing ascorbic acid. With
known concentration of iodine, the concentration of ascorbic acid can be found through concepts of
titrations where the number of moles of the titrant equal the number of moles of the analyte present in
solution. Using this fact, the concentration of ascorbic acid can be found for both the whole fruit and
orange juice. It should be noted that iodine is easily evaporated at standard atmospheric pressure and
temperature so iodine concentrations may vary over time. To limit this error, the bottle provided by lab
was always kept closed and parafilm was used to cover the buret.

The iodine concentration was found using known amounts of 100% ascorbic acid in lab. From
the mass of the ascorbic acid used, we may determine the moles of ascorbic acid present in solution.
Using the iodine, we may reach the end point, the point at which the solution reacts with the starch to turn
from a colorless to a blue color. This color will signify the exact mole equivalence of the analyte with the
titrant. Using the volume of iodine titrant used to reach the end point, the concentration of iodine is
7
found .

The ascorbic acid solution was made using a 250.00 mL volumetric flask which is calibrated at
20° Celsius. The normal room temperature was not accounted for and would need to be accounted for in
future experiments. In addition, the 250.00 mL volumetric flask has a tolerance associated with it of ±0.12
7
mL . This tolerance should be noted for the calculation of the concentration of iodine for future
experiments as well.

There is known error that is present from the instruments used during this experiment. Before
handling the buret, the buret is primed with iodine solution and expelled 3 times as waste to ensure that
all contaminants in the buret are washed out. After loading the buret, the next biggest error to avoid is
buret reading. To minimize the error of reading the buret wrong, a buret reader was fashioned to
increase the accuracy of reading the buret volume. In addition, error can be introduced if the buret is not
7
read at the same height as one’s eyes known as parallax . To avoid parallax, one’s eyes should be at the
same level as the meniscus. The meniscus is where the measurement should be read because it
represents the true volume that is not affected by surface tension of the inside of the buret. The inside of
the buret is also not made precisely straight which can lead to small errors due to a variance in buret
7
width . To minimize this error, the buret was filled around the 25 mL mark for all titrations. The 50 mL
buret will also have a small tolerance of ±0.05 mL during reading. To prepare for the Iodometric titration,
7
air bubbles must be checked inside the buret and near the stop cock to ensure accurate results . If air
bubbles are present, it may affect the titration by up to 3 drops of titrant because the buret reading is
accounting for the air bubble. The speed at which the liquids are expelled during the titration should also
7
be noted because the iodine solution is very viscous in nature . The tendency of the viscous iodine to
stick to the walls is high and can introduce error if the titration occurs too rapidly. After all titrations were
finished, the buret was rinsed with deionized water and placed upside down. All titrations occurred with
the same buret for each experiment and trial.

The titration itself must be carefully monitored because the stir bar used can spray drops of
analyte onto the 250 mL Erlenmeyer flask. Deionized water was used to spray the sides to release the
analyte from the walls of the flask and into the main portion of the solution. This ensures that all the
analyte is being accounted for during the titration.

Volumetric pipets were used to deliver 25.00 mL of the ascorbic acid solution made in lab also
has tolerances associated with it. For this experiment, we used a 25.00 mL volumetric pipet which as a
7
tolerance of ±0.03 . This tolerance can lead to small amounts of error when factoring in the reading error
of a volumetric pipet though it is more precise than the buret. The volumetric pipet was also primed 3
times before each experiment with the analyte used. In this case, the ascorbic acid solution made in lab
was used to prime the volumetric pipet.

Once these errors have ben accounted for, the iodine average and confidence interval given by
the experiment, the solution of iodine was determined to be 0.0107 (±0.001) M iodine at 95% confidence.
The three trials had an RSD of below 10 parts per thousand and an interesting observation happened if
you change the decimal places for the RSD calculation. When inputting the values for the RSD
calculations, the number of decimal places used changes the RSD value greatly. The RSD can vary up
to ±2 parts per thousand depending on the number of decimal places used. The confidence interval is
very low and suggests that the iodine solution should be within ±0.001. It was found that the iodine
concentration for the batch was supposed to made to 0.011 M iodine. This would suggest that the value
determined by the Iodometric titration of known amounts of ascorbic acid overlaps with the expected
value of 0.011M.

Orange juice
 The analysis of ascorbic acid in Tropicana Pure Premium Orange Juice was done to determine
the milligrams of ascorbic acid in 8 ounces of Tropicana Pure Premium Orange Juice. Using the same
concepts as the determination of iodine concentration, the volume of iodine expelled into the titrant at the
end point is the stoichiometric equivalence of the moles of titrant to the moles of analyte. Now, the iodine
concentration was found to be 0.0107 M iodine and can be used to solve for the concentration of ascorbic
acid. The concentration of ascorbic acid was converted using dimensional analysis to usable units of
milligrams of ascorbic acid in 8 ounces of the Tropicana orange juice.

The juice supplied was always on ice when measured to ensure the freshness of the orange juice
and its ascorbic acid content. There can be some indeterminate error associated with the raising
temperature of the orange juice as it sits on the lab bench for use. Every student in the class data would
have tested the orange juice at different temperatures until it reaches the room temperature. There raises
a concern with the ascorbic acid and whether or not it degrades as the temperature raises. The juice was
expelled using the same 25.00 mL volumetric pipet which was primed 3 times before use in the
experiment to reduce the contaminant level inside the volumetric pipet. Again, the same errors
associated with the buret and the volumetric pipet are going to apply to this determination of ascorbic acid
as well.

It was found that the mean of the ascorbic acid content in orange juice was found to be 109.31
(±3) mg of ascorbic acid per 8 ounces of fruit juice. The confidence interval is considerably low and
suggests that the true value is within 3 mg of ascorbic acid per 8 ounces of fruit juice of the mean 109.31.
The expected value can be first located by the nutrition label where the vitamin C content is 120% in 8
8
fluid ounces of the daily value of a 2000 calorie diet . The FDA states that the daily value, 100%, of
8
vitamin c in a 2000 calorie diet is 60 mg of vitamin c . Therefore, 120% of daily value for 8 fluid ounces
would be 72 mg of vitamin c in the 8 fluid ounces. There is a big difference between the mean
determined experimentally, 109.31 (±3) mg of ascorbic acid per 8 ounces of fruit juice, and the expected
value of 72 mg of ascorbic acid per 8 ounces of fruit juice. This difference in mean is due to the strict
laws that are imposed on food manufacturers where the nutrition label states the minimum amounts of
nutrients found on the nutrition label. In this case, the minimum milligrams of ascorbic acid are 72 mg per
8 ounces of fruit juice. The mean, 109.31 (±3) mg of ascorbic acid per 8 ounces of fruit juice, is above
this threshold and therefore a valid value.

There is a great deal of error associated with this the finding of the end point. It is difficult to get
the end point exactly at the end point which should be considered. The orange color of the orange juice
make sit difficult to compare the end points compared to the known ascorbic acid solution which was
colorless. This, in itself, is an error that would also lead to comparison errors of the class data. There
was also bias in the titration from the user error but was reduced by only blindly measuring the values and
not calculating to see the differences until all trials were done. In this way, it eliminates the bias towards a
theoretical mean and prevents working around a theoretical mean. In addition, the titration was done a
different day and would be interesting to see if the iodine concentration changed over the course of time
being exposed to the air. To eliminate this error, finding the concentration of the iodine on the same day
given the same conditions would have been ideal but there was not enough time in class to complete this
task in addition to the task at hand.

Whole Fruit

The determination of ascorbic acid in whole fruit was done by the same procedures outlined in
the experimental and results section. The oranges, obtained by Ralph’s Grocery Store, was juiced the
same day it was purchased using the juicer available in lab. The juice was strained of any pulp to get a
pure liquid of orange juice because the pulp would change the amount expelled from the volumetric pipet.
A big error to avoid would be to strain the orange juice a few times to ensure that no pulp is within the
fresh orange juice. Again, the concentration of ascorbic acid in the whole fruit can be done because of
the known iodine concentration measured in previous sections.

It was found that the whole orange juice contained 42.53 (±0.8) milligrams of ascorbic acid per 1
fruit. The confidence interval was very small when compared to the mean and suggests that the true
value is within ±0.8 mg of ascorbic acid per 1 fruit. The expected value for a standard medium orange is
70 mg ascorbic acid in 1 fruit. There is a differences in the mean determined experimentally, 42.53 (±0.8)
9
milligrams of ascorbic acid per 1 fruit, and the 51 mg of ascorbic acid in 1 fruit . The differences between
the two values is most likely due to the age of the orange. The orange was purchased the same day it
was juiced but it was not known how long it was when it was brought to the store. One of the biggest
differences was size and weight of the fruit where 1 medium orange did not specify a certain weight, size,
or age. The oranges were bought the same day but it does not mean that the oranges were from the
same batch grown in the same location. It seems that the USDA value is an average of all oranges sold
in the United States of an average weight and size. Knowing this, the experimentally determined value
was not consistent with the expected value.

Comparison of Data Collected to Class Data of All Burets

The comparison of iodine concentrations collected in the results section with the other 15 burets
was done to see if the data collected in this experiment was an outlier. Shown in table 1, The F test gave
7
an Fcalculated value of 2874 which is enormously high compared to the Ftable value of 19.5 . This suggest
that the standard deviations of the two data sets were very different from each other. This result was
expected because the standard deviation of the class data was large because of the different amounts of
people running the experiments. Within each student, the standard deviation is most likely low but for the
class as a whole, each student’s standard deviation would differ from one another leading to a larger
-5
standard deviation. The standard deviation in this experiment was found to be 3.86 x 10 M iodine which
is smaller than the standard deviation found from the class, 0.00207 M iodine. The standard deviation
from the molarity of iodine is seen to be 3 times in magnitude greater tan the standard deviation found in
this experiment. This finding can also be due to each student’s standard for the end point was different.
Though most student’s used the same experiment article run by the University of Canterbury to compare
6
the end point of values, each student’s perception to the end point is different .

Since the standard deviations of the iodine concentrations with data collected in the experiment
compared against the class data was calculated, the means for each data set was performed using a T
test. The means for the iodine concentration for the data collected in this experiment was 0.0107 M
iodine. The class data average for the iodine concentration was found to be 0.0117 M iodine. Using
these means allowed us to calculate the Tcalculated which was found to be 0.891 as seen in table 1. This
value was compared to the Ttable value of 2.021 using 40 degrees of freedom. Since the tcalculated value is
7
less than the Ttable value listed in Harris , we can say that the means of the data sets are not statistically
different. This suggest that the data collected in this experiment was not statistically different from the
class data. With these results, the iodine concentration collected in this experiment was shown to be not
statistically different and, therefore, acceptable to use in the comparison of burets.

The same procedure was used for the analysis of ascorbic acid in 8 ounces of orange juice. The
standard deviation found during this experiment was found to be 1.11 milligrams of ascorbic acid per 8
ounces of juice. When compared to the class data, excluding my data, the standard deviation was found
to be 11.73 milligrams of ascorbic acid per 8 ounces of fruit juice. Using the F test, the Fcalculated value
was found to be 125 as shown in table 1. The Fcalculated value is much lower than that of the iodine
7
concentration. The Ftable was found to be the same value as the iodine concentration, 125 . In this case,
the Fcalculated value is greater than the Ftable value which suggests that the standard deviation of each data
set is statistically different. Again, this is most likely due to the differences in the class to perceive their
end point compared to others’ end point. With everyone’s end point being slightly different, it is expected
to see a large standard deviation and, as a result, a large Fcalculated value.

The mean for the ascorbic acid per 8 ounces of fruit juice collected in this experiment was also
compared to the mean of the class data to verify if the data collected in this experiment can be used for
further comparisons. The mean for the milligrams of ascorbic acid per 8 ounces of fruit juice found in this
paper was 109.31 milligrams of ascorbic acid per 8 ounces while the mean for the class data was 119.90
milligrams of ascorbic acid per 8 ounces. The means of each seem to be statistically different but the T
test reveals that the Tcalculated is 1.55 as seen in table 1. The Ttable value was 2.021 which is still greater
than the Tcalculated value. As a result, the Tcalculated value was smaller than the Ttable value which shows that
the means are not statistically different.

This further suggests that the data collected in this experiment was no different in terms of
accuracy, due to having statistically identical averages. Though, the F test shows that the standard
deviation for each data set was different suggests that the precision of the instrument is different. This
may also be due to the the differences of perceived end point. This suggests that there is no difference in
terms of accuracy of the class data including my data. This allows for the comparison of each buret
according to size and comparison of different sized burets because the validity of each person’s
technique lead to the same iodine concentration and orange juice concentration. In addition, the iodine
concentration was found to overlap with the 0.011 M iodine given in class with a 1.17 (±0.004) M Iodine
experimentally found using an Iodometric titrations. The orange juice also overlaps the minimum ascorbic
acid content in a 8 ounce serving with 119.89 (±10) milligrams of ascorbic acid in 8 ounces of fruit juice
experimentally found.

Comparison of Data Collected to Class Data Holding Buret Size Constant

Since the data collected in this experiment was not statistically different from the class data, the
comparison of the data with the buret of the same diameter. The large burets were compared to the other
7 large burets in the class. The standard deviations for the concentration of iodine were found to be
-5
3.86 x 10 M iodine for the large buret that was used in this experiment. The class data standard
deviation was found to be 0.00240 M iodine. Using these standard deviations, the F test revealed
Fcalculated value was 3581 where the Ftable value was found to be 19.0 as seen in table 2. This shows that
the standard deviations are significantly different when comparing the data collected in this experiment
with the data collecting from the other 7 large burets. Much like the last comparison, there is a
discrepancy between the standard deviations because of each person’s perceived end point. Though
each student might have similar standard deviations individually, when comparing each student to
another student their standard deviations will differ greatly.

The means for the iodine concentration with the data collected from this paper compared to the
other 7 large burets were found to be not statistically different. The means for the iodine concentration for
this paper was found 0.010649 whereas the iodine concentration for the other 7 burets was found to be
0.010748 as shown in table 1. The T test shows that the tcalculated was found to be 0.070 which is
extremely low compared to the ttable value shown as 2.086. This shows that the means are not statistically
different when comparing burets of the same size.

The concentration of ascorbic acid per 8 ounces of orange juice was determined for the data
collected in this paper with the other 7 large burets in class. The standard deviation calculated for the
data in this experiment was 1.05 milligrams of ascorbic acid per 8 ounces of fruit juice while the standard
deviation for the other 7 large burets was found to be 3.79 milligrams of ascorbic acid per 8 ounces of fruit
juice. These standard deviations would undergo the F test and reveal an Fcalculated value of 13 shown in
table 2. Comparing this value to the Ftable value of 19.4 shows that the Fcalculated value is smaller than the
Ftable value, suggesting that the two standard deviations for the large burets were not statistically different.
This is a different result from the last experiment where the orange juice concentration of ascorbic acid
per 8 ounces of fruit juice were statistically different in terms of standard deviations. This result shows
that the standard deviations were not different and may be explained by having the students with the
large burets sitting all next to each other. This allows for the comparison of the end point with each other
by looking at the end point of nearby students. In the comparison of the the class data to all burets, the
end points may differ because of the location of the small burets. I would expect that the comparison of
the small burets to another small buret would have the same results with a small Fcalculated value.

The means for the milligrams of ascorbic acid per 8 ounces of fruit juice was compared between
the data collected on the large buret with the 7 other large burets. The mean for the data collected in this
paper was found to be 109.31 milligrams of ascorbic acid per 8 ounces. The mean found for the other 7
large burets was found to be 113.00 milligrams of ascorbic acid per 8 ounces. According to the T test,
the tcalculated value was shown to be 1.65 which is smaller than the ttable value listed in table 2, 2.021. Since
the tcalculated value was smaller than the ttable value, the means for the milligrams of ascorbic acid in 8
ounces of fruit juice are not statistically different between the large burets.

Comparison of Large Burets to Small Burets

The large burets were then compared to the small burets on the basis of precision and accuracy.
It is hypothesized that the small burets will have greater precision and thus better accuracy due to its
small diameter. Since the titration is only done with 5 to 10 mL, it is hypothesized to be advantageous to
use a buret with a small volume capacity and a smaller diameter. The smaller diameter would result in a
large increase in measurability for a small volume expelled for the titration. The larger buret has a larger
volume capacity which is also not needed for this experiment.

The iodine concentration for the large buret was found to be 0.0107 M iodine with the standard
deviation of 0.0223 M iodine. For the smaller buret, the iodine concentration as found to be 0.0126 M
iodine with a standard deviation of 0.00126 M iodine. The standard deviations were subject to a F test
which would see if the standard deviations are significantly different. The Fcalculated, shown in table 3, was
found to be 3.177 for the iodine concentration which was greater than the Ftable value of 2.12. Since the
Fcalculated value was larger than the Ftable value, the standard deviations are statistically different. This
would suggest that the level of precision for measuring the iodine concentration is different for the burets.
Since the larger burets have been used and considered precise, the smaller burets are statistically
different and considered less precise.

The average iodine concentration from the large burets and the small burets were compared
using a T test. The mean iodine, as stated, was found to be 0.0107 M iodine for the large buret and
0.0126 M iodine for the small buret. These means were subject to a T test which showed a tcalculated of
3.53 seen in table 3. Comparison to the ttable value of 2.766 shows that the tcalculated is greater than the ttable
value. This suggests that the means are statistically different. This would reject the hypothesis that the
smaller buret would have greater precision with greater accuracy because of the F test and T test.
Because the F test had to be rejected, it was shown that the precision was different as well as the
accuracy was different The true value for the iodine concentration was set to be 0.011 M made in the
same batch. The average for the larger buret had a greater accuracy receiving an average of 0.0107 M
iodine. The smaller burets showed an increased amount from the true value with a value of 0.0126 M
iodine.

The large burets and small burets ability to measure the milligrams of ascorbic acid in 8 ounces of
orange juice was also compared. The standard deviations for the large buret was found to be 3.76
milligrams of 8 ounces of orange juice whereas the standard deviation for the smaller buret was found to
be 13.02 milligrams of ascorbic acid per 8 ounces of orange juice. These values were placed into table 4
for direct comparison of the large burets and the small burets. The standard deviations were subject to
7
the same F test and it was shown to have an Fcalculated of 12.00 with a Ftable value of 2.12 . This suggests
that the standard deviations are statistically different and suggests that the precision of the two
instruments is different. We may say that the larger buret had greater precision than the smaller burets
due to having a smaller standard deviation.

There is some error that can be associated with the Iodometric titration of the orange juice
because of the position of the endpoint. Those grouped in the large burets might have agreed upon one
end point because of the proximity of each large buret student. The students with the small burets were
not in view of the large buret so the small buret students may have agreed upon the color of the end point
based on their proximity to one another. These difference of end point determination can result in both
the increase and decrease of the iodine and orange juice concentrations. The iodine concentration may
have less indeterminate error due to being a colorless analyte, though, the orange juice may prove to be
more difficult to determine the end point.
Table 4: Comparison the large buret and small buret based on iodine concentration (M) and orange juice
(mg/8oz). There is an increase in the iodine concentration when using the small burets which travels
further away from the expected iodine concentration of 0.011 M iodine. The orange juice concentration
was also greater than the orange juice concentration when performed in the large buret.

Type of Buret Iodine Concentration (M) Orange Juice (mg/8oz)

Large Buret 0.0107 (±0.005) M 112.54 (±8) mg/8oz
Small Buret 0.0126 (±0.003) M 125.46 (±30) mg/8oz

The T test would reveal if there is a difference in the accuracy of the large buret compared to the
smaller buret for the measurement of ascorbic acid in 8 ounces of fruit juice. The large buret had a mean
of 112.54 milligrams of ascorbic acid in 8 ounces of fruit juice with a standard deviation of 3.76 milligrams
of ascorbic acid in 8 ounces of fruit juice. The small buret had a larger mean value of 125.46 milligrams
of ascorbic acid in 8 ounces with a larger standard deviation of 13.02. Using the means, the Tcalculated
showed 4.68 for the milligrams of ascorbic acid per 8 ounces of fruit juice which was greater than the ttable
given, 2.021. This would show that the means for determination of ascorbic acid in orange juice were
statistically different. Thus, one buret size Is more accurate than the other buret. The Tropicana 100%
Pure Orange Juice states that it has a daily value of 120% for an 8 ounce glass of fruit juice in a 2000
8
calorie diet . As explained in an earlier section, there are at least 72 milligrams of ascorbic acid in an 8
9
ounce serving of fruit juice . Both values from the large buret and small buret are valid in that they are
above the 72 milligrams of ascorbic acid threshold. Though, the large buret have been used for this
experiment countless times in the past and are considered the most accurate form of performing this
experiment. The hypothesis was that introducing the small buret would have effects in better precision
and increased accuracy. According to this T test, the smaller buret is not suitable to replace the larger
buret because it was not considered accurate compared to the larger buret by having a larger tcalculated
value compared to the ttable value.

For future experiments, It is important to note that using the large buret will give more precise and
accurate measurements for both the iodine concentration and ascorbic acid content in orange juice. The
small burets should not be used to replace the larger burets because of this. I think to better gauge the
end point, every student should have their first trial checked to see if they have reached the end point and
come to a whole class agreement what the end point should be. Comparing the end point to the article is
difficult because of the differences on computer screens.

Conclusion

The determination of ascorbic acid through Iodometric titration is possible and was performed
using whole fruit and Tropicana Orange Juice. Once the iodine concentration was found using known
amounts of ascorbic acid, the Iodometric titration of whole fruit and Tropicana Orange Juice was done. It
was found that the iodine solution provided in lab was 0.0107 (±0.001) M Iodine. This number overlaps
with the expected value of 0.011 M iodine that the laboratory technicians prepared before the class
started. Using this, the milligrams of ascorbic acid in the Tropicana Orange Juice was shown to yield
109.31 (±3) mg of ascorbic acid per 8 ounces of Tropicana Pure Premium Orange Juice. The expected
value for the orange juice was found to be 72 mg of ascorbic acid per 8 ounces of Tropicana Pure
Premium Orange Juice. This difference in found means is due to that value being the minimum amount
of ascorbic acid present in the 8 ounce serving. In which case, the value that was experimentally found
was deemed correct. The Iodometric titration performed on a whole fruit showed that a whole fruit
contained 42.53 (±0.8) milligrams of ascorbic acid per 1 fruit which was near the expected value of 51 mg
of ascorbic acid per fruit. The comparison of the data collected in this experiment was compared to the
class data. It showed that the precision of the data was different but the accuracy was the same for both
the iodine concentration and the orange juice ascorbic acid per 8 ounces. A similar pattern emerged
when the comparison of data collected in this experiment compared to those with the same buret size.
For the iodine concentration, there was a difference in standard deviations but the accuracy was similar of
those of the same buret size. Though, for the orange juice there was no difference in precision nor
accuracy as shown in the F and T tests. For the comparison of large and small burets, there was a
difference in the precision and accuracy for both the iodine concentration and the milligrams of orange
juice for 8 ounces of orange juice. Therefore, the small burets cannot replace the large burets for future
experiments.
 References

(1) Voet, Donald, Judith G. Voet, and Charlotte W. Pratt. Fundamentals of Biochemistry: Life at the
Molecular Level. Hoboken, NJ: Wiley, 2013. Print.

(2) https://www.nlm.nih.gov/medlineplus/ency/article/000355.htm

(3) http://www.webmd.com/drugs/2/drug-322/ascorbic+acid+oral/details

(4) https://ods.od.nih.gov/factsheets/VitaminC-Consumer/

(5) Haan, John. Experiments for Quantitative Analysis Laboratory Fullerton: Montezuma Publishing,
2015. Print.

(6) University of Canterbury College of Science. "Determination of Vitamin C Concentration by

Titration."

(7) Harris, Daniel C. Quantitative Chemical Analysis. 8th ed. New York: W.H. Freeman, 2010. Print.

(8) http://www.fda.gov/Food/GuidanceRegulation/GuidanceDocumentsRegulatoryInformation/Labelin
gNutrition/ucm064928.htm

(9) http://ndb.nal.usda.gov/ndb/foods/show/2284?fgcd=&manu=&lfacet=&format=&count=&max=35&
offset=&sort=&qlookup=09200