The Concentration of Citric Acid in

Low Acid Orange Juice Versus Regular Orange Juice

Nathan Orlando - Katelyn Parker

Macomb Mathematics Science Technology Center

Chemistry

10A

Mrs. Hilliard / Mr. Supal / Mrs. Dewey

May 24 2017

Table of Contents
 Orlando - Parker

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

Review of Literature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

Problem Statement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

Experimental Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Data and Observations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Data Analysis and Interpretation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Appendix A . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Appendix B . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Appendix C . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Works Cited . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Introduction

Companies today are coming out with alternatives to allow people to enjoy their favorite

foods and drinks, but with less acid. One of the most common drinks that has been modified, is

orange juice. Orange juice is naturally high in acid, due to the citric acid in citrus fruits. The low

acid orange juice is advertised to be more useful for people with acid reflux.

The purpose of this research was to find whether low acid orange juice actually had a

lower citric acid content than that of regular no pulp orange juice. It was hypothesized that the

low acid orange juice would not have a lower citric acid concentration, but that the citric acid

concentration of the regular no pulp orange juice and the low acid orange juice would be equal.

From the data collected from the acid-base titration of orange juice and sodium hydroxide 0.1M

solution, the equivalence point of each type of orange juice were calculated, and then used in a

statistical test to determine whether low acid orange juice actually had a lower citric acid content

than that of regular no pulp orange juice.

Through these research results, it is hoped that people can make smarter decisions when

choosing low acid alternatives to orange juice. Acid reflux and enamel breakdown are a common

issue between people when drinking orange juice. Acid reflux is caused by the acid in the orange

juice raising the pH in the stomach. As the pH increases, the stomach allows acid to raise into the

esophagus causing a burning pain. The acid also erodes enamel which exposes the micro tubules

containing nerves in the teeth. This then leads to sensitive teeth and adds pain when eating and

drinking certain foods or beverages. This research allows people who are affected by acidic

foods or beverages to determine for themselves whether there is too much acid in the low acid

orange juice or not.

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Review of Literature

Orange juice is a very popular thing to drink especially with breakfast. Many Americans

believe that orange juice is helpful in a healthy diet. But, orange juice can be just as harmful due

to the amount of citric acid in orange juice. Citric acid in the orange juice will erode the enamel

in your teeth (“Ouch”) and can cause acid reflux. The purpose of this research was to find the

concentration of citric acid in regular no pulp orange juice and compare it to the citric acid

concentration of low acid orange juice to see whether or not low acid orange juice actually has a

lower concentration of citric acid, as advertised.

Based off of popularity, Tropicana™ was the brand of orange juice chosen. This makes

the experiment applicable to many areas because the juice is utilized by large amounts of people.

Tropicana make a low acid orange juice as well as an original no pulp. These will be titrated to

see if there is actually a difference in citric acid content. If there is no difference, the orange juice

is being falsely advertised. Also, the consumers that were looking for juice that would not give

them heartburn or other issues would still obtain these issues due to the false advertising.

High acid content can cause health problems to the consumer. First off, acid erosion may

occur. This is the wearing away of enamel in the teeth. Sensitive, yellow teeth is an effect of this

because dentin, hard bony tissue underneath the enamel, is exposed. (“Ouch”)

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Figure 1. Tooth Diagram

Under the dentin are microscopic channels known as tubules. These tubules lead to the

pulp in the center of teeth which contains the nerves and blood vessels. Figure 1, above, shows a

detailed diagram of a tooth. When someone has sensitive teeth, drinking eating hot, cold, sweet,

acidic, or spicy foods and beverages will cause pain (“Ouch”).

Orange juice is a beverage that can trigger acid reflux symptoms because of the large

amounts of citric acid found in it. For anyone that has acid reflux disease, also known as

gastroesophageal reflux disease (GERD), this is a major issue when consuming acidic beverages.

This is because the acid raises the pH in your stomach and acid reflux occurs as an attempt to

lower it. Some symptoms of acid reflux include coughing, nausea, hoarseness, belching, sore

throat, and regurgitation (Robinson). Acid reflux occurs because the ring of muscle leading into

the stomach, also know as the lower esophageal sphincter (LES), is supposed to completely close

after a food or drink passes through it. If the LES does not close all the way then acid is allowed

to travel up into the esophagus and cause pain and or discomfort (Robinson). Acid reflux can

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occur even when the consumer does not have GERD. Figure 2 shows a diagram of the stomach

and gives a better understanding on how acid reflux actually occurs.

Figure 2. Acid Reflux

The main acidic component of many juices is citric acid (C6H8O7). This acid is found in

citrus fruits, like oranges. It is also used as a preservative and in sour candies. The concentration

of citric acid in the orange juice was found by using acid-base titrations. Acid-base titration is a

technique used in chemistry to find the concentration of a solution by reacting it

stoichiometrically with a solution of known concentration until it has completely reacted. The

endpoint is reached when the indicator, a compound that changes in color based on the pH of its

surroundings, which is added to the acid being titrated, changes color. This signifies the end of

the reaction, and thus, the concentration of the unknown solution can be found. Not all indicators

are correct for every substance though. Each indicator has a different pH range, or is used to

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indicate a certain thing in the substance.

Figure 3. Titration Setup

Figure 3 above shows the setup for a titration. The burette holds the titrant, and the

conical flask, or erlenmeyer flask, holds the analyte. During a titration, the titrant is dripped into

the analyte to titrate the analyte. Not shown in the above picture is the pH sensor, which gives a

real time graph of the pH, known as a pH curve (“Acid-Base Titration Curves”). In this research,

the orange juice was the analyte, or the solution of unknown concentration, and the titrant, or the

solution with a known concentration, was sodium hydroxide (NaOH). Sodium hydroxide is a

strong base, while the acid in orange juice, citric acid , is a weak acid.

Figure 4. Sodium Hydroxide And Citric Acid Neutralization Reaction

Shown in the above figure is the molecular formula for neutralizing sodium hydroxide

and citric acid. When the two react during the titration, the sodium hydroxide neutralizes the

citric acid. The concentration of the citric acid is found by measuring how much sodium

hydroxide (in liters) was used to reach the end point, or the complete neutralization of the citric

acid, and this is then multiplied by the molarity of sodium hydroxide to calculate the number of

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moles of sodium hydroxide that were required to reach the end point. After the number of moles

is found, they can be plugged into the mole ratio of 1 mole of citric acid to 3 moles of sodium

hydroxide. This is used to find the moles of citric acid, which is then used to find the mass of

citric acid in the orange juice. The mass of the citric acid is found by multiplying the number of

moles of citric acid by its molar mass. The concentration of citric acid in the juice can finally be

found by putting the mass of the citric acid over the volume of the orange juice used multiplied

by 100 mL. It is multiplied by 100 mL because the concentration of acid in juices is normally

expressed in grams of acid per 100 mL of juice.

Previous research, conducted at the University of the Philippines Los Banos, used

pineapple juice and calamansi juice, rather than orange juice, but still determined the

concentration of citric acid in each juice to find which juice was more acidic (Madrid). Both

juices were titrated using sodium hydroxide. The sodium hydroxide titrated the citric acid via

acid-base neutralization, which means that the sodium hydroxide reacted with the citric acid until

it was completely neutralized.

Performing this experiment required an understanding of many chemistry topics.

Apprehending titrations as well as an assimilation of how acidic drinks affect teeth and acid

reflux is critical to conducting the experiment and comprehending how the results can be applied

to the real world. Data from this research will be utilized to help people with general dental

problems and people with acid reflux decide if low acid orange juice is a better option than

regular orange juice. This data can help people make better health decisions when it comes to

orange juice, and in turn help people live more comfortably with acid reflux or dental issues.

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Problem Statement

Problem:

The purpose of this experiment was to determine if low acid orange juice actually has a

lower citric acid concentration compared to regular orange juice. The results of this experiment

will help decide which orange juice has less effect on the consumer’s teeth enamel and acid

reflux symptoms. This will help limit tooth enamel erosion and pain from acid reflux.

Hypothesis:

It was hypothesized that the concentration of citric acid in the 20 ml of low acid content

orange juice will not be lower than the concentration of citric acid in regular orange juice, but

rather the same citric acid concentration.

Data Measured:

The independent variable of this experiment was the type of orange juice that was

titrated. Low acid orange juice and regular orange juice were the two types titrated. The

constants were the amount of pulp and the brand of the orange juice. The dependent variable was

the concentration of citric acid in each type of orange juice, measured in moles per liter. To

analyze the data and its significance, descriptive statistics were used.

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Experimental Design
Materials:

pH probe 300 ml No pulp Tropicana

orange juice
(0.1M) 1000 ml Sodium hydroxide (NaOH)
Phenolphthalein indicator
300 ml Low-acid Tropicana orange juice
Pipet
Vernier labquest 50 ml graduated
cylinder
200 ml Beaker Goggles
1000 ml Distilled water Small stirring
bar
Magnetic stirrer

Procedures:

1. Obtain and wear goggles.

2. Set up the titration according to Appendix A.

3. Turn on the labquest and set it up according to appendix B.

4. Take the burette off of the holder and hold it in one hand over a small beaker, and place a

funnel into the top. Make sure that the burette is closed, and then pour in sodium

hydroxide solution until the burette is full to the 0 ml line. If it goes over, open the

stopcock and let out sodium hydroxide solution until it is at the 0 ml line.

5. Check the tip for air bubbles. If there are air bubbles, open the stopcock and as the

solution flows, gently whack the side of the tip to release the air bubble. Once this is done

close the stopcock and empty the solution out of the small beaker.

6. Record the initial amount of sodium hydroxide in the burette before beginning the

titration. Make sure to read the burette at eye level, at the bottom of the meniscus, for the

most accurate reading.

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7. Using a large beaker, place in the small stirring bar, then measure out 20 ml of orange

juice into the large beaker. Put in four drops of phenolphthalein indicator.

8. Place this underneath the prepared burette, on top of a magnetic stirrer.

9. Turn on the magnetic stirrer, and let the stirring bar adjust into a steady and sure stir

before beginning.

10. Start the data collection on the Labquest by pressing the play button. Hit keep and type in

0 ml to record the beginning pH.

11. Begin titrating by turning the stopcock and adjusting it into a steady drip. Watch the pH

on the Labquest closely, and hit keep and record the burette reading at every 0.5 pH

increase. Be sure to close the stopcock before recording the data in the labquest.

12. Repeat step 12 until the graph on the Labquest has created an S shaped curve.

13. Record the end amount of sodium hydroxide in the burette, and save the pH curve to

analyze.

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Diagram:

Figure 5. Materials Used

The diagram above shows all materials used in this experiment. The burette is also setup

and the labquest is ready to record data. The white bottle with a dropper is the phenolphthalein

indicator. Both types of orange juice are shown as well as the 0.1 molarity sodium hydroxide

solution, shown between the orange juices.

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Data and Observations

Table 1 below shows the data calculated for the regular orange juice titrations.

Table 1
Concentration of Citric Acid in Regular Orange Juice
Amount of
Orange Amount of Calculated
Equivalence Juice Equivalence Moles Moles of Orange Juice Concentration of
Point Titrated Point of Citric Titrated Citric Acid
Trial (ml) (ml) (L) NaOH Acid (L) (M)
1 20.4 20 0.0204 0.00204 0.00068 0.02 0.03

4 21.2 20 0.0212 0.00212 0.00071 0.02 0.04

5 20.6 20 0.0206 0.00206 0.00069 0.02 0.03

6 21.2 20 0.0212 0.00212 0.00071 0.02 0.04

8 21.2 20 0.0212 0.00212 0.00071 0.02 0.04

9 19.2 20 0.0192 0.00192 0.00064 0.02 0.03

13 20.1 20 0.0201 0.00201 0.00067 0.02 0.03

14 19.7 20 0.0197 0.00197 0.00066 0.02 0.03

15 21.9 20 0.0219 0.00219 0.00073 0.02 0.04

16 20.4 20 0.0204 0.00204 0.00068 0.02 0.03

18 19.8 20 0.0198 0.00198 0.00066 0.02 0.03

19 20.5 20 0.0205 0.00205 0.00068 0.02 0.03

20 19.1 20 0.0191 0.00191 0.00064 0.02 0.03

25 19.7 20 0.0197 0.00197 0.00066 0.02 0.03

28 19.7 20 0.0197 0.00197 0.00066 0.02 0.03

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Table 1 above has only slight differences in the concentration of citric acid, switching

between 0.03 to .04. The equivalence point was calculated using the first derivative function in

the Logger Pro software, shown in figures 4 and 5.

Figure 6. Balanced Chemical Equation For Sodium Hydroxide and Citric Acid

Figure 6 above shows the balanced chemical equation for sodium hydroxide and citric

acid. This equation was used to obtain the mole ratios needed to convert the amount of titrant

used from liters to moles. The ratio is 1 mole of citric acid to every 3 moles of sodium

hydroxide. This ratio is used in the formula shown in figure 2.

Figure 7. Formula and Sample Calculation For Moles of Citric Acid

Figure 7 above shows the formula and a sample calculation for calculating the moles of

citric acid. The equivalence point in liters was calculated by multiplying the number of moles of

NaOH by 0.1 moles over 1 liter, because the solution used was a 0.1M solution of sodium

hydroxide. This was then multiplied by the mole ratio of 1 mole of citric acid over 3 moles of

sodium hydroxide. The numbers for the mole ratio were gained from the balanced chemical

equation shown in figure 6.

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Figure 8. Formula and Sample Calculation For Concentration of Citric Acid In Orange Juice

The formula shown above in figure 8 calculates the concentration of citric acid in orange

juice, otherwise known as the molarity. To calculate the concentration of citric acid in orange

juice, the moles of citric acid were divided by the amount of orange juice titrated in liters. The

amount of orange juice titrated in liters was calculated by multiplying the amount of orange juice

titrated in milliliters by one one-thousandth.

Figure 9. Trial 1 Data Points

In Figure 9, one of the thirty trials’ data points are shown. A similar table was done for

each trial, but with thirty trials, not all could be shown. In the above table, the D1 column is the

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1st derivative column. The 1st derivative tells the slope of a tangent line to the curve at any

instant. The peak of the derivative column is the equivalence point, which is when the moles of

sodium hydroxide solution added are equal to the moles of citric acid in the orange juice.

Figure 10. Trial 1 First Derivative Graph

Figure 10 above shows the first derivative graph for trial one, which was a regular orange

juice trial. The equivalence point is 20.4 ml because at that point in the titration, the moles of

citric acid and the moles of sodium hydroxide solution were equal.

Table 2
Concentration of Citric Acid In Low Acid Orange Juice
Amount of
Orange Amount of Amount of Calculated
Equivalence Juice Titrant Moles Moles of Orange Juice Concentration
Point Titrated Used of Citric Titrated of Citric Acid
Trial (ml) (ml) (L) NaOH Acid (L) (M)
2 18.2 20 0.0182 0.00182 0.000607 0.02 0.03033

3 18.4 20 0.0184 0.00184 0.000613 0.02 0.03067

Amount of
Orange Amount of Amount of Calculated
Equivalence Juice Titrant Moles Moles of Orange Juice Concentration
Point Titrated Used of Citric Titrated of Citric Acid
Trial (ml) (ml) (L) NaOH Acid (L) (M)
7 18.8 20 0.0188 0.00188 0.000627 0.02 0.03133

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10 19.3 20 0.0193 0.00193 0.000643 0.02 0.03217

11 19 20 0.0190 0.00190 0.000633 0.02 0.03167

12 20 20 0.0200 0.00200 0.000667 0.02 0.03333

17 18 20 0.0180 0.00180 0.000600 0.02 0.03000

21 17.5 20 0.0175 0.00175 0.000583 0.02 0.02917

22 18 20 0.0180 0.00180 0.000600 0.02 0.03000

23 18.5 20 0.0185 0.00185 0.000617 0.02 0.03083

24 19.6 20 0.0196 0.00196 0.000653 0.02 0.03267

26 18.1 20 0.0181 0.00181 0.000603 0.02 0.03017

27 19 20 0.0190 0.00190 0.000633 0.02 0.03167

29 18.1 20 0.0181 0.00181 0.000603 0.02 0.03017

30 19.7 20 0.0197 0.00197 0.000657 0.02 0.03283

Table 2 above shows the calculated data for the low acid orange juice titrations. The

equivalence point was calculated as discussed above under figure 4. The concentration of citric

acid for these trials were all around 0.03, with only slight differences with the numbers below

them.

Table 3
Observations For Low Acid Orange Juice Titrations
Trial Low Acid Observations

2 Partner 1 ran this trial. Took a very long time because it was the first trial. Some
readings were a little off.

3 Partner 1 ran this trial. No observations were written down for this trial. Final buret

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reading seems to be higher than most other low acid trials.

7 Partner 1 ran this trial. Small black particles in the orange juice. Bubbles in the buret
made readings difficult. NaOH was splashing onto the side of the beaker. Color
changed to a orange at around 8.75 pH. All points are near perfect pH wise.

10 Partner 1 ran this trial. NaOH hitting sides of the beaker. Initial reading was a little off.
Point 6 was messed up and deleted. Point 7 is 0.1 above where it should be. Color tint
changed at 9.2 pH 20.1 NaOH.

11 Partner 1 ran this trial.Small amounts of water left in the beaker before adding sodium
hydroxide to the juice.

12 Partner 2 ran this trial.

17 A different person ran this trial because Partner 2 was not there this day.

21 Partner 2 ran this trial.

22 Partner 2 ran this trial. This trial ran smoothly, with no data collecting problems.

23 Partner 1 ran this trial. Seems to have a larger amount of citric acid compared to most
other low acid trials.

24 Partner 2 ran this trial. The phenolphthalein indicator color started to change at a pH of
around 8.1.

26 Partner 1 ran this trial. Points are a little off. No observations written.

27 Partner 1 ran this trial. Points are a little off. No observations written.

29 Partner 1 ran this trial. Color change at 9 pH. Points are all near perfect. Trial ran great.

30 Partner 2 ran this trial on the labquest. This was one of the best trials, but there were a
few points that were off.
Table 2 above has the observations for the low acid orange juice titrations. The data

collection was observed as slightly off for many trials.

Table 4
Observations For Regular Orange Juice Titrations
Trial Regular Orange Juice Observations

1 Partner 2 ran this trial. This was the first trial ran, and took awhile to set up. The data
points for this trial were recorded differently than the others due to a
miscommunication. They were recorded every .05 pH change rather than every .5 pH
change.

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4 Partner 2 ran this trial. This trial went faster than the first one, but still a little slow
because partner 2 was adjusting to the procedure and changing the experimental design
to fit the process of the titration better as she ran her titration.

5 Partner 1 ran this trial. Missed point during the trial. Otherwise, it went well.

6 Partner 2 ran this trial. This trial ran well and quick, with nothing unusual to record.

8 Partner 2 ran this trial. At about 9.3, the color changed to red. One point in the middle
of the s curve was input mistakenly, causing a little blip in the curve.

9 Partner 2 ran this trial. This trial ran smoothly, there were no blips in the curve.

13 Partner 1 ran this trial. Small black particles in orange juice. Points are a little off. Lots
of missed points as well as off points. Color change at around 9 pH.

14 Partner 1 ran this trial. Point 7.4 pH is 0.1 over what I was looking for. Missed point at
8.3. Missed point at 10.8.

15 Partner 2 ran this trial.

16 Partner 1 ran this trial. Point 6.2 pH at 6.33 instead. Strikethrough 20.00 supposed to be
20.4.

18 Partner 2 ran this trial.

19 Partner 1 ran this trial. Skipped a point at 7.7. Skipped a point at 9.7 pH.

20 Partner 2 ran this trial. Nothing out of the ordinary happened.

25 Partner 2 ran this trial. Nothing unusual happened.

28 Partner 2 ran this trial. Nothing unusual happened. The trial went a little quicker.

Table 4 above shows the observations from the regular orange juice titrations. Some of

the trials had trouble, and many of the trials had points that were off by .1 to .4.

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Data Analysis and Interpretation

This experiment involves titrating two types of orange juice. One low acid and one

regular. Fifteen trials of each type were tested. They were tested by adding NaOH from a burette

to the juice. This allowed proper measurements of how much NaOH was used. After measuring

the amount of NaOH used to get the equivalence point, the amount of citric acid could be

calculated. The unit for this calculation is molarity. In the end, a lower citric acid concentration

in the low acid orange juice is desired.

Multiple steps were taken in this experiment to make sure that accurate data was

collected from each titration. First, the titrations needed to be repeated properly. Fifteen trials of

both types of orange juice were recorded. This helped prove the normality of the data as well as

assure that there were no outliers. Next, each trial must be randomized. The order in which each

trial was conducted was determined by calculating a random integer between 1 and 2. If 1 was

chosen, a low acid trial would be conducted next. If a 2 was chosen, a regular orange juice trial

would be conducted next. Finally, every trial was performed under the same conditions in the

same testing environment so certain lurking variables will be the same for all trials.

The statistical test that best suits the data is a Two-sample t test along with Descriptive

Statistics because it shows if two averages are significant or happened due to chance alone. This

can be applied because this experiment uses two types of orange juice. A Two-sample t test

determines if two independent samples have significant averages or averages that occurred due

to chance alone. One of the two assumptions were met for this statistical test. These were having

independent samples and at least thirty trials completed and or a normal distribution according to

the Central Limit Theorem (CLT). Due to time constraints, only fifteen trials of each type of

orange juice were run. Before running the two sample t-test, the normality of the data must be

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verified. To do so, the data can be either graphed as box plots, or as normal probability plots. In

this research, both were done to ensure the normality of the data.

Figure 11. Box Plots of Citric Acid Concentration For Both Orange Juice Types

Figure 11 above shows the boxplots of the citric acid concentrations for both types of

orange juice. The low acid orange juice box plot is slightly left skewed because the median of

0.03083 is closer to the 1st quartile than the 3rd quartile. This could affect the normality of the

data, so to further check the normality of the data, normal probability plots were made, and are

shown in figures 12 and 13. The regular orange juice box plot is relatively normal because the

median is approximately in the middle of the box plot. By looking at these box plots, it can be

observed that there is an overlap of the higher 50% of the low acid orange juice citric acid

concentrations, and the lower 50% of the regular orange juice citric acid concentration. This

could mean that the data is not statistically significant, but at the same time, the medians are 50%

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away from each other, which shows that the data could be statistically significant. The two

sample t-test will be effective in correctly determining the statistical significance of the data.

Figure 12. Normal Probability Plot of Regular Orange Juice Citric Acid Concentrations

Figure 12 above shows the normal probability plot for the citric acid concentrations of the

regular orange juice. The data is assessed as normal because the data points follow the same

pattern and stick close to line, showing that the data points that do not stick closer to the line and

that aren’t spaced out too much are normal.

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Figure 13. Normal Probability Plot of Low Acid Orange Juice Citric Acid Concentration

Figure 13 above shows the normal probability plot for the low acid orange juice citric

acid concentration. This plot shows that the data is relatively normal because the data points are

close to the line, and follow the line, rather than being spread out and further away from the line.

Because the data is relatively normal, a statistical test assumes the data will be normal can be run

to determine whether the data is statistically significant or not.

Figure 14. Null And Alternative Hypotheses

Figure 14 above shows both the null and the alternative hypotheses. μ1 stands for the

average citric acid concentration of the regular orange juice, and μ2 stands for the average citric

acid concentration of the low acid orange juice. The null hypothesis, Ho, is that the means of both

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populations, the average concentration of the regular orange juice and the average concentration

of the low acid orange juice, are the same. The alternative hypothesis is that the average citric

acid concentration of the regular orange juice is greater than the average citric acid concentration

of the low acid orange juice.

Figure 15. Two Sample T-Test Data

Figure 15 above shows the data calculated from running a two sample t-test. How to

calculate the t value and The t value is 5.78488, which means that the t value is almost 6 standard

deviations above or below 0. With the t value being this great, there is a strong chance that the

null hypothesis will be rejected. The p value of 0.000002 is less than the alpha level of 0.05, so

therefore, the null hypothesis can be rejected. With that, the alternative hypothesis is accepted, so

therefore, there is a statistically significant difference within the data. This means that there is

evidence that the citric acid concentration of regular orange juice is higher than the citric acid

concentration of low acid orange juice. If the null hypothesis was true, there is between a .05%

and a 0% chance of getting a difference between the means of 0.00272 by chance alone.

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Figure 16. Two Sample T-Test P graph

Figure 16 above shows the p graph for the two sample t-test that was run. The p value

was 0.000002. This value was plotted onto the p graph, but the graphing is not visible because

the value was so low that it went to the far left of the tail. The t value, which is the standard

deviations away from the mean is not shown on this graph because it is above the 4 and -4 range

of the graph. When the p value is plotted onto the graph, it shows that there is a 0.000002 chance

on both sides of 0 that there will be a difference observed from the null hypothesis that is at least

as extreme as the difference in the data, assuming that the null hypothesis is true. The total sum

of the probability is 0.000004. What this means is that when the null hypothesis is true, the t-

value falls within these shaded regions .0004% of the time. This means that the null hypothesis

can be rejected, because the p value of 0.000002 is below the alpha level of .05.

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Conclusion

The purpose of this research was to find whether or not orange juice advertised as low

acid had a lower concentration of citric acid than regular no pulp orange juice. The hypothesis

was that the concentration of citric acid in low acid orange juice would not be less than that of

the regular orange juice, but rather that the two citric acid concentrations would be the same.

This hypothesis was rejected, because the concentration of citric acid in the low acid orange juice

was lower than the concentration of citric acid in regular no pulp orange juice. The null

hypothesis was also rejected. This shows that there is a significant difference in the acid

concentrations of both types of orange juice.

Not all assumptions were met by the experiment. Only 15 trials of each type of orange

juice were performed. This means that there was not enough data collected to ensure it is normal.

Descriptive statistics was used to show the normality of the data. Figures 12 and 13, in the data

analysis, show normality plots of both the low acid and regular citric acid concentrations. These

plots show the data to be relatively normal which allows a two-sample t test to be calculated.

After finding the t and p values for the experiment, it was concluded that regular orange juice has

a significantly higher amount of citric acid than low acid orange juice. This is because the p

value of 0.000002 was lower than the alpha level of 0.05. Therefore, the null hypothesis can be

rejected. In the end, this shows that there is evidence that the concentration of citric acid in

regular orange juice is higher than the concentration of citric acid in low acid orange juice.

The hypothesis was not supported by the data because citric acid was not only in the

regular orange juice, but was also used as a preservative to keep it from spoiling quickly. This

extra citric acid contributed to the overall results of the research. Citric acid was used as a

preservative because citric acid lowers the pH of the overall solution, causing uncomfortable

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living conditions for the enzymes that over ripen fruit and vegetables. This causes their growth to

slow down and can kill them, helping to keep the food fresh longer. In the low acid orange juice,

a calcium salt of citric acid, calcium citrate, (Ca3(C6H5O7)2 was used as the preservative. This

was used rather than citric acid to keep a lower concentration of acid, because it has a pH of 5.6

to 5.8, rather than the lower pH of citric acid. Calcium citrate was also used rather than other

preservatives because it adds the same flavor as citric acid, but without the high acid content. To

get the data needed to conclude that the hypothesis was rejected, an acid-base titration of the

orange juice and sodium hydroxide was run. In figure 17 below, the chemical equation for the

citric acid in orange juice and sodium hydroxide is shown. An acid-base titration is when an acid

or a base of a known concentration is used to neutralize an acid or a base of an unknown

concentration.

Figure 17. Chemical Equation of Sodium Hydroxide and Citric Acid

In Figure 17 above, the sodium hydroxide neutralizes the citric acid by raising the pH.

This is done by a molecule of the base, sodium hydroxide, accepting a proton from a molecule

of the acid, citric acid. The proton is a hydrogen atom, and the negative log of the concentration

of the hydrogen atoms in a solution is the pH. By taking away the hydrogen atoms from the citric

acid, it raises the pH, because there is a lower concentration of hydrogen atoms in the orange

juice solution, and that is just for each molecule. Adding drops of the sodium hydroxide to the

orange juice causes large scale reactions between the molecules because there are thousands of

molecules just in each drop.

The experimental design contributed in different ways to the experiment. There was a

negative effect on it from improper steps. Many changes had to be made to it so it would ensure

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better data collection. Therefore, throughout the data collection the experimental design was seen

as flawed and was not trusted. This limited the use of it until it was changed. The change

happened after a few trials. This ensured the the rest of the trials were repeated correctly. Then,

the amount of NaOH used to titrate each type of orange juice was around the same. Visually,

since the experiment design requested the use of an indicator, the color change was seen to

happen around the same point. Overall, it had a more positive effect on the conclusion because it

helped infer that the regular orange juice has a significantly higher amount of citric acid.

Many mistakes were made throughout the experiment. These were mostly with the data

collection, such as when data points were missed during collection, or when the stopcock would

be turned too far and solution would start squirting out rather than falling into the orange juice at

a steady drip. Another mistake that was made was that the orange juice was left out for long

periods of time during the research, which could have caused it to go slightly bad, and could

have affected the results. Ways to minimize these mistakes are to plan ahead early, and be aware

that the first trial will take awhile because there is a lot to set up. To minimize missing data

points, a steady and slow drip of sodium hydroxide into the solution would make it more

efficient to stop the drip and record a point, rather than a fast drip. To minimize the error of

solution squirting out, it must be remembered that the stop cock is touchy, and the slightest turn

changes the drip very quickly. It’s best to start out with a slow drip and gently speed it up to an

appropriate speed, rather than to start dripping quickly or squirt out the solution.

An alternate experiment that finds which acidic beverage is the best for people with acid

reflux would be interesting. Since any beverage can be titrated, there is a ton of room to change

or expand this experiment. The data found from performing these experiments could prove that a

company or brand is advertising incorrectly. Also, it shows the public which beverage suits their

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condition. For example, this experiment tested how much citric acid is in low acid orange juice.

This is important for people to know because acid reflux is common and someone will want to

drink orange juice without triggering anything. All in all, this experiment is extremely flexible

and can allow people who struggle with acid reflux or have Gastroesophageal Reflux Disease to

chose a beverage that does not trigger anything.

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Appendix A: Titration Setup

Materials:

Burette (0.01 ml precision) Burette clamp

Ring stand 200 ml Beaker
Stir plate

Procedure:

1. Grab the burette clamp, and slide it onto the ring stand from the top of the pole.

2. Set it at an appropriate height, and then tighten it using the screw in the middle.

3. Attach the burette by pushing it between the three . Make sure that the burette is

vertically straight, to assure a good reading.

4. Twist the burette so that the numbered side is readable, then tighten the clamps onto the

burette.

5. Check that the stopcock is closed at the bottom of the burette. If the lever on the side of

the stopcock is vertical, it is open. If it is horizontal, then it is closed.

6. Place a 200 ml beaker beneath the burette so that if any access liquid is released before

the titration begins, it will not fall into the orange juice or onto the magnetic stirrer.

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Diagram:

Figure 18. Titration Setup

Figure 18 shows a setup of a titration. The burette is twisted so that it is readable. The pH

probe is located as far left on the beaker as it can go so it does not get in the way of the titrant. It

is also connected to the labquest which is recording the pH. This is all setup on a stir plate with a

magnetic pill so the pH can be recorded quickly and accurately.

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Appendix B: Setup of Vernier Labquest

Materials:

Vernier Labquest pH probe

Flash drive Utility clamp
Vernier labquest charger

Procedure:

1. Turn on the Vernier Labquest, then connect the pH probe to the Labquest.

2. Plug in the charger to ensure that it will not die during the trial.

3. Place the flashdrive into the Labquest for later.

4. Choose New from the File menu.

5. Change the mode to event based entry, and name the event Amount of NaOH and input

the unit as milliliters.

6. Use a utility clamp to attach the pH probe to the ring stand, then place it into the orange

juice. Adjust it so that it is near the edge so as to not hit the stirring bar.

Diagram:

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Figure 19. Labquest Setup

The Figure above shows a completed graph of a titration. Each point is placed with an

increment of 0.5 pH. But, there are two missing points in this example. As you can see from the

graph, the equivalence point is around the pH of 9. This will vary between experiments.

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Appendix C: Sample Calculations

To analyze the data the following equation was used. This is the Two-sample t test

equation that will calculate the t value for the experiment. x̅1 is the mean concentration for the

regular orange juice. x̅2 is the mean for the low acid orange juice citric acid concentrations. s1

stands for the standard deviation of the regular orange juice citric acid concentrations, and s2

stands for the standard deviation of the low acid orange juice citric acid concentrations. n1

stands for the number of trials in the regular orange juice citric acid concentrations, and n2 stands

for the number of trials in the low acid orange juice citric acid concentrations.

Figure 20. Two Sample t - Test Formula and Sample Equation

The equation above calculated the t value which represents the number of standard

deviations above or below the mean on a t distribution. The t value is used to find the probability

as a range of percents that the sample data will be found again by chance alone, if the null

hypothesis were true. To find the range of percents, a t distribution critical values chart is used.

The degrees of freedom are found by subtracting one from the smallest sample size, so for this

research it would be 14. Find that number on the left side, then follow the row corresponding to

it down until two numbers that the t value is between are found. Once that is found, the columns

are followed up, and the numbers at the top as percents are the range of percents. If, like in this

research, the t value goes further than the chart, the range of percents will be from 0% to the last

number at the top of the chart as a percent.

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Works Cited

"Acid-Base Titration Curves." Khan Academy, https://www.khanacademy.org/test-

prep/mcat/chemical-processes/titrations-and-solubility-equilibria/a/acid-base-titration-

curves. Accessed 15 Mar. 2017

Bhatia, Manish. "Hyperacidity, Acid Reflux, Esophagitis & Peptic Ulcers." Hpathy, Sept.

2009, hpathy.com/homeopathy-papers/hyperacidity-acid-reflux-esophagitis-peptic-

ulcers/. Accessed 15 Mar. 2017

Daniel, Smriti. "Guarding Against GERD." Mediscene, 17 Oct. 2010,

www.sundaytimes.lk/101017/MediScene/mediscene_3.html. Accessed 15 Mar. 2017

Jolliffe, Diane M. "Practical Gastric Physiology." Oxford Academic, 1 Dec. 2009,

https://academic.oup.com/bjaed/article/9/6/173/378337/Practical-gastric-physiology.

Accessed 15 Mar. 2017

Lloyd, William C. "Sensitive Teeth." healthgrades,

https://www.healthgrades.com/symptoms/sensitive-teeth. Accessed 15 Mar. 2017

Madrid, Ariel J. "Determination of The Acidity of Fruit Juices." Academia,

https://www.academia.edu/9259242/Determination_of_the_Acidity_of_Fruit_Juices.

Accessed 15 Mar. 2017

"Ouch, My Sensitive Teeth!." Family Health Online,

www.familyhealthonline.ca/fho/dental/DH_sensitiveTeeth_FHa08.asp. Accessed 15 Mar.

2017

Robinson, Jennifer. "What Is Acid Reflux Disease." Web MD,

www.webmd.com/heartburn-gerd/guide/what-is-acid-reflux-disease#3. Accessed 15 Mar.

2017

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