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Introduction…………………………………………………………………....…………1

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

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

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

Conclusion……………………………………………………………………………...26

Acknowledgements……………………………………………………………………30

Appendix C: Randomization…………………………………………....…………….34

Works Cited………………………………………....………………………………….37

Introduction

Fournier - Lay - Weidemann

According to Marc Silver, of npr.org, 153.6 Liters of soda per capita are

consumed in the United States. The acids these beverages have within can have

a long term detrimental effect on tooth enamel. This experiment was designed to

test how much these acids actually affect teeth by examining how eggshells, a

formidable substitute for teeth, would hold up against phosphoric acid and citric

acid. Mass loss helps show the harmful effects that acids have on the calcium in

teeth, and can be used to determine the effects when a reaction occurs between

Using the experimental design and Two Sample t-Tests, the results of the

people can learn how to take better care of their teeth and doctors may even be

able to find a way to counteract the effects of acids on tooth enamel. The acids

molarities were set to be as close to the molarity of the acids found within the

liquid. The mass is measured before and after soaking the eggs in these acids,

subtracting them to find the mass loss, then this data is put through a Two

Review of Literature

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Acids are well known for their ability to break down substances and their

atomic structure. Many of the foods and beverages that society consumes

One of the acids being tested is citric acid, the most prominent acid

present in orange juice. Citric acid has a pH around 2.2, proving it is strong

Livestrong.com, juices contain a high amount of acid and sugar, which creates a

Phosphoric acid (H3PO4) is one of the most common acids in soda pop.

Soda pop is one of the most popular drinks worldwide. Phosphoric acid has a

non-constant pH level, but the most common pH level found for it is around 1.1.

This means that this substance is very acidic and can easily wear down teeth

soda pop have 10 times more of an effect on teeth than the acids found in fruit

juice do.

Eggshells and teeth both consist of a large quantity of calcium. According to Bill

and eggshells are composed similarly, they both contain a calcium base.

Eggshells are made out of calcium carbonate (CaCO3), and a tooth is composed

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composites, have similar rigid structures made out of tiny mineral like crystals. In

order to use the eggshells, the yolk and the egg whites must be siphoned out.

The erosion of the teeth will be measured in grams. The change in mass will

show how much the eggshells eroded. This idea was found in an experiment by

the Journal of Zhejiang University. In their experiment, they used the units of

It’s very easy to tell when teeth are eroding. According to Delta Dental

from Smileway, some ways to notice teeth erosion are: the teeth are more

sensitive, the teeth become discoloured, the teeth get a bit more rounded, the

edges of the teeth begin to look more transparent, the teeth will begin to crack,

and small dents may appear on the chewing surface of the teeth. These methods

will not work for the experiment, therefore the before and after weight of the

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This image portrays how acids can decay teeth and where it makes the

biggest impact. The acid trapped underneath the plaque begins dissolving

enamel. The plaque continuously builds up when it is not cleaned off properly,

capturing more acid and helping the enamel dissolve. The gum level recedes and

exposes the crown more, and gives the acid more space to degrade the enamel.

Acid breaks down enamel by leaching the calcium from the teeth.

bacteria in the mouth. This bacteria uses sugar from foods and drinks to produce

acids that dissolve and damage the teeth. It was stated in a journal by Nutrition

Research, that dental erosion is the leaching of minerals from the tooth structure.

Acid is able to remove minerals from a tooth structure using a process called

mineral ions that can only exist in liquid solutions. In the presence of acids

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millions, and even billions, of calcium and other mineral ions are removed from

hydroxyapatite latticework of teeth. Due to the loss of these ions the teeth will

This image shows how the process of demineralization occurs when teeth

are involved. As seen in the image the minerals in the teeth are leached away by

A study by the Journal of Dentistry was done to test the effects of acidic

food and beverages on teeth by leaving the teeth in the food. Diluted acids will be

used on the eggshells to show how the concentration of acids within the

Another study by PLoS ONE tested the effects of these foods on the teeth

for a term of 7 days. Due to the time constraints of this experiment, the eggshells

will not be in the acids for this period of time. In their experiment the acids were

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Studies by other groups have all addressed similar problems to the one

that will be discussed in this paper. The design of this experiment will come from

support the results by documenting the changes in appearance after the teeth

are placed in the acids. The recent studies provide background information on

Problem Statement

Problem Statement:

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The purpose of this experiment was to test the effects of Phosphoric Acid

(H3PO4) and Citric Acid (C6H8O7) on the decay rate of eggshells (CaCO3). The

results of this experiment helped determine the effects that acids can have on

enamel and teeth, when they have been exposed to acid for a long period of

time.

Hypothesis:

Acid, the Citric Acid will cause the eggshells to decay at a greater value.

Data Measured:

Phosphoric Acid and Citric Acid. The control group of this experiment was to

leave the eggshells sitting in water, this is because there is no way to tell if there

is truly a change in the eggshells mass without comparing it to just having the

eggshells sit out in air or water (H2O). The constants of the experiment was 200

mL of Phosphoric Acid, Citric Acid, and water. The dependent variable of the

experiment was the change in mass, in milligrams, of the eggshells over the

duration of the experiment. To analyze and test the significance of this data, a

Two-Sample T-Test was used to compare the difference of mass between the

acids and water. Descriptive analysis was then used to talk about the results

Experimental Design

Materials:

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4L of 0.055 M Phosphoric Acid 500mL Graduated Cylinder

(H3PO4) 4L of 0.047 M Citric Acid (C6H8O7)

Scale (0.0001g Precision) 35 of 532mL Red Solo Cups

1 Weigh Boat 3 Tongs

1000 mL Beaker 1 Scupula

Figure 3. Materials

In the figure above, the materials needed to conduct the experiment are

Procedure:

1. Measure the initial mass of the eggshell in grams using a scale.

2. Fill a red solo cup with 200 mL of 0.055 M Phosphoric Acid solution.

3. Fill another red solo cup with 200 mL of 0.047 M Citric Acid solution.

5. Set an eggshell in each of the red solo cups. Be sure to completely fill egg

so it will not float.

6. Remove eggshell from solution using tongs and allow shell to sit 2 days to

dry.

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Data:

Table 1.

Change of Mass in Eggs Soaked in Phosphoric Acid

0.055 M Phosphoric Acid

Initial Mass Final Mass Change

Trial # (g) (g) (g)

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2 6.1221 5.1542 -0.9679

3 6.7002 5.6988 -1.0014

4 6.7564 6.1646 -0.5918

5 6.1960 5.7849 -0.4111

6 5.9466 5.5646 -0.3820

7 6.1726 5.5006 -0.6720

8 5.6350 5.4696 -0.1654

9 6.2393 5.3641 -0.8752

10 6.4739 5.3914 -1.0825

11 5.3911 5.0732 -0.3179

12 5.4011 5.7951 0.3940

13 6.4989 5.7199 -0.7790

14 6.6070 5.1417 -1.4653

15 6.4261 5.6770 -0.7491

Average Mass Loss: -0.7055

figures. This was determined by how majority of the data can go up to 4 decimal

place accuracy, due to the scales. As shown above, Trial 12’s data is not similar

that the cause of this error could have been due to the lack of crystallization

Table 2.

Change of Mass in Eggs Soaked in Citric Acid

0.047 M Citric Acid

Initial Mass Final Mass Change

Trial # (g) (g) (g)

16 6.8617 5.0487 -1.8130

17 6.8283 5.0909 -1.7374

18 7.8853 4.4112 -3.4741

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20 6.0306 4.9658 -1.0648

21 5.1908 4.0915 -1.0993

22 5.7519 4.1610 -1.5909

23 6.5050 4.7382 -1.7668

24 5.9310 4.5295 -1.4015

25 6.2012 4.6217 -1.5795

26 6.3346 4.4676 -1.8670

27 6.1152 4.9486 -1.1666

28 6.5591 4.3330 -2.2261

29 6.5324 4.8253 -1.7071

30 6.1099 4.4405 -1.6694

Average Mass Loss: -1.6932

This data above shows no outliers within the data set. It is also at a

significant figure count of 5, due to the scales precision. This data shows one

outlier, Trial 18. This egg most likely lost majority of its mass because of it’s lack

of membrane and the top of the egg almost being completely gone.

Table 3.

Change of Mass in Eggs Soaked in Water Control

0 M Distilled Water

Initial Mass Final Mass Change

Trial # (g) (g) (g)

31 6.5791 6.2593 -0.3198

32 6.7160 6.3590 -0.3570

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34 6.9408 6.5630 -0.3778

35 6.6264 6.1432 -0.4832

Average Mass Loss: -0.4029

The control group is meant to determine if there was a true change when

another factor acted upon it. This average will be used to determine the mean of

the mass lost from the acids, by subtracting the water average from it.

Observations:

Table 4.

Observations of the 15 Eggs Soaked in Phosphoric Acid

0.055 M Phosphoric Acid

Trial # Observations

1 Lead Markings, Tiny Bottom hole, Medium Top Hole

2 Lead Markings, Tiny Bottom hole, Medium Top Hole

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4 Even Medium Holes on Top and Bottom

5 Large Top Hole, Small Bottom Hole

6 Small Top Hole, Tiny Bottom Hole

7 Large Top Hole, Small Bottom Hole

8 Large Top Hole, Small Bottom Hole

9 Small Top Hole, Tiny Bottom Hole

10 Medium Top Hole, Small Bottom Hole

11 Large Top Hole, Small Bottom Hole

12 Small Top Hole, Tiny Bottom Hole, Less Crystallization

13 Medium Top Hole, Tiny Bottom Hole

14 Small Top Hole, Tiny Bottom Hole

15 Small Top Hole, Tiny Bottom Hole

Throughout the experiment, all of the eggs that were in the Phosphoric

Acid appeared to have crystallization forming on the outer shell. Shown in the

data, Trial 12 appeared to have less crystallization, showing a lack of work from

Table 5.

Observations of the 15 Eggs Soaked in Citric Acid

0.047 M Citric Acid

Trial # Observations

16 No Cracks, Most Residue Wiped, Small Hole on Bottom.

17 No Cracks, Most Residue Wiped, Small Hole on Bottom.

18 No Membrane, Big Crack, Residue Left Over, Tiny Hole on Bottom

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Some Cracks near Top Base, Almost All Residue Wiped, Medium

20 Bottom Hole

Cracks on Top and Bottom Base. Medium Hole on Bottom, Slight

21 Residue Left Over

No Cracks, Medium Top Hole and Small Bottom Hole, Residue

22 Mostly Wiped

23 No Cracks, Lots of Residue, Small Bottom Hole, Medium Top Hole

24 Inner Membrane Smashed, Top is Concave, Small Bottom Hole

25 Crack on Top, Small Top, Medium Bottom Hole, Some Residue

26 Crushed in Half, Few Residue, Holes unseeable

27 Crack Through Bottom Hole, Small Bottom Hole, Slight Residue

28 Medium Holes, Slight Residue, No Cracks

29 Small Holes, No Cracks, Slight Residue

30 Giant Top Hole, Small Bottom Hole, No Cracks, Slight Residue

All of the eggs that had gone into the citric acid had foam surrounding it,

hence the residue in the observations. Before weighing these eggs, the residue

Table 6.

Observations of the 5 Eggs Soaked in Water Control

0 M Distilled Water

Trial # Observations

31 Large Top Hole, Tiny Bottom Hole

32 Medium Top Hole, Tiny Bottom Hole

33 Large Crack on Top, Lead Markings on Top

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35 Small Top Hole, Tiny Bottom Hole

All of the eggs in the control group were relatively normal eggs, the only

issue being the water the eggs were in became murky. Although the murkiness

seems to have no effect on the egg, it is still worth noting. The other issue with

the water eggs is that all of the inner membranes were missing, showing a mass

loss.

The test being used to evaluate data in this experiment is a Two Sample

are essential to this test. These three assumptions are: there are two simple

random samples from two distinct independent populations, both samples are

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from populations that are normally distributed, and both samples are no more

than one tenth their populations. Due to there being fifteen trials per treatment, a

normal probability plot must be used to determine if the populations are normal, if

Figure 4 shows the normal probability plot for the data collected on the

eggs soaked in Phosphoric Acid. The data collected on Phosphoric Acid appears

follows the line closely. Due to the fact that Trial 12 is the only egg to gain weight

it is considered an outlier.

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Figure 5 contains the normal probability plot for eggs soaked in Citric Acid.

Due to the fact that the points form a loose line they can be considered relatively

normal. However, there is one outlier at 3.4741 grams lost, trial 18. This outlier

does not follow the plot, and may make the data appear to be less normal. After

testing with, and without the outlier. It was determined that it did not affect the

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Figure 6 contains the normal probability plot for the water control group.

The points on the plot follow the line loosely. This gives the data an appearance

of being relatively normal, as most of the plots follow the line snugly.

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As shown, the red box plot, Citric Acid, has an outlier. The blue box plot,

Phosphoric Acid, does not contain an outlier, but in the data there is still one

number that is unlike the rest. This number was the trial in which the egg gained

weight. This plot provides evidence that Citric Acid was the most effective

treatment on the eggs, because it had a higher median than the other solutions.

The Phosphoric Acid appears to be normal in the box plot, but the other two

appear to be skewed. The Citric Acid box appears to be very slightly skewed to

the left. This may be due to more eggs losing more weight. The water plot

appears to have a slight right skew which may be due to a larger number in the

data. This may have happened due to the fact that only 5 eggs were used in this

group, although this group only exists to test if the acids had an actual effect.

𝐻𝑜: 𝜇1 = 𝜇2

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𝐻𝑎: 𝜇1 < 𝜇2

Figure 8. Null and Alternative Hypothesis between Phosphoric and Citric Acid

Figure 8 shows the null and alternative hypothesis that will be used to

carry out the two sample t-Tests that will be carried out. The symbol μ is used to

mean of citric acid. The null hypothesis states that the sample means are equal

to each other. The alternative hypothesis states that the citric acid had a greater

Figure 9 shows the data that is included in the Two Sample t-Test. These

symbols include the “PVal” which is the p value of the test. This is found by using

the “t” value and finding the corresponding p value on Table C. The standard

these are 15 because there were 15 trials each acid. After calculating this the t

value was found to be -4.97536. Using Table C, the p value was determined. The

p value is the probability of obtaining results that are equal to or more extreme

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than what was actually observed when the null hypothesis is true. The p value of

this test was determined to be 0.000016. The fact that the p-value is so much

lower than the alpha level provides strong evidence against the null hypothesis.

Figure 10 shows the chart to show the p-value of the data. As seen in the

image it does not indicate where the p-value would fall. This is the case because

the p-value is so small that it will not appear on the chart. The smallest p-value

that this chart can hold is 0.0001, but the p-value was 0.000016. Proving that

The null hypothesis is rejected due to the fact that the p value is less than the

alpha level of 0.05. There is significant evidence that the citric acid dissolved

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more of the egg. This means that there is less than a 0.05 chance of getting

𝐻𝑜: 𝜇1 = 𝜇2

𝐻𝑎: 𝜇1 < 𝜇2

Figure 11. Null and Alternative Hypothesis for Water and Citric Acid Groups

Figure 11 shows the null and alternative hypothesis for the relation

between the citric acid and the distilled water. The null hypothesis is used to

check if the data is equal to each other. The alternative hypothesis is used to test

if the citric acid has more of an effect than the water. Once again μ is used to

represent the populations of both the citric acid and the distilled water. The water

is represented by μ1, and the citric acid is represented by μ2. The null hypothesis

states that the mean mass lost for the two populations are even. The alternative

hypothesis states that the mean mass lost for the citric acid is greater than the

Figure 12. Data from T-Test Involving Water and Citric Acid

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Figure 12 shows the data that was used to conduct the Two Sample t-

Test. The means are represented by the x-bar one and x-bar two. X-bar one

represents the water and x-bar two represents the citric acid. The standard

deviations of the data are represented by sx1 and sx2. N1 and N2 are used to

represent the populations of the samples. These were used in the formula to find

the t-value. After performing the calculations it was found to be that the t value

was -8.29747. After finding the t value then p-value was found using Table C.

After using Table C the p-value appeared to be around 2.46878 x 10-7. Due to

the

fact that the p-value is so low this is strong evidence against the null hypothesis.

Figure 13. P Chart for the Effect of Water and Citric Acid

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Figure 13 shows the chart of the water and citric acid effects. Just as in

Figure 8 the p-value is so small that it is not visible on the chart. The p-value is

The researchers reject the null hypothesis due to the fact that the p-value is

much lower than the alpha level of 0.05.There is significant evidence that the

citric acid dissolved more of the egg shell than the water did. This means that

there is a very low chance that results this extreme will happen by chance alone.

𝐻𝑜: 𝜇1 = 𝜇2

𝐻𝑎: 𝜇1 < 𝜇2

Figure 14. Null and Alternative Hypothesis for Phosphoric Acid and Water

Figure 14 shows the null and alternative hypothesis for the effect of the

phosphoric acid and water. In this hypothesis, μ1 represents the mean of the

weight lost to water, and μ2 represents the mean of the weight lost to phosphoric

acid. The null hypothesis states that the two means would be even, and the

alternate hypothesis says that the mean loss of phosphoric acid is greater than

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Conclusion

phosphoric acid had the ability to erode the tooth substitute, eggshells, when

exposed for a period of forty eight hours. A Two Sample t-Test was used to test

the significance of the experiment, and compare the sample means of the three

different experiment groups - citric acid, phosphoric acid, and a water control.

Descriptive statistics were then used to visually represent the results found. The

hypothesis of the experiment predicted that the citric acid would have the most

significant effect on the eggshells. Due to the results of the Two Sample t-Test, it

was determined that the citric acid had the most significant impact on the

eggshells therefore, the hypothesis was accepted. The solution that had the least

significant effect was the water control, this proves the effectiveness of it working

as a control, because it helps show the lack of effect that water has on teeth.

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The mean mass lost from each of the treatments was analyzed. The eggs

soaked in citric acid had an average mass loss of approximately 1.6932 grams.

The phosphoric acid had an average loss of 0.7055 grams, and the water had an

average loss of 0.4029 grams. The mean of each acid needs the mean of the

water constants to be removed to determine the true loss from the acid, rather

than the water that diluted it. The citric acid lost significantly more mass than the

rest of the egg groups, as it lost 0.9877 grams more on average than the

phosphoric acid group, and lost 1.2903 grams more on average than the water

groups. The p-value that compared the sample means between the citric and

significant at the alpha level of 5% and 1%. The p-value comparing the sample

mean of the citric acid to the water was approximately 2.46878x10 -7, roughly 0%,

which was also significant at the alpha level of 5% and 1%.These p-values show

The sample size did not meet the requirements of a Two Sample t-Test,

where 30 data points are required, but was deemed normal by using the normal

distribution test. All three normal distributions plots suggest that all of the data

was relatively normal, allowing the use of a Two Sample t-Test. The box plots

also displayed a degree of normality with only the water appearing to have a

slight skew. However, there was an outlier in the citric acid group. The position of

the box plots also supported the hypothesis. The citric acid box was farthest to

the right, proving it had the most effect on the eggshells. The phosphoric acid

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was in between the water and the citric acid, and the water had the least effect

This evidence does not support the research that states that the

phosphoric acid should have more of an effect on the eggshells than the citric

acid. This is due to the fact that the acid breaks down enamel by removing the

calcium, weakening the structure. Some reasons this could have occurred is that

the molarities may have been slightly inaccurate. Also, the short time period

could have caused the eggs to not have enough time to fully achieve the most

accurate results. If the eggs would have sat longer, it is possible that the

outcome of the experiment could have been different. The longer the exposure

the eggs have to the acids the more opportunity the acids have to leach calcium

from the structure of the tooth. In this experiment, the data was found that citric

acid, at a 0.047M, did more damage than phosphoric acid, at a 0.055M. This

goes against the research this experiment was based off of. Due to the data

using a substitute for teeth, the results may be slightly inaccurate to how the

acids work on actual teeth. The phosphoric may have more of an effect on actual

tooth enamel, but the citric acid was shown to have a higher effect on the

eggshells.

The experimental design was important for this experiment. It allowed the

would not be mixed up when they were drying. The experimental design also

allowed the experimenters to work productively, and work through the lack of

time provided. One problem with the experimental design is that sometimes a

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certain solution would run out and an experimenter would have to make more of

the solution while the others were working. This could have affected the

Mistakes were made throughout the experiment such as the hole size in

the egg made it float above the water. Another mistake was that some of the

eggs broke when they were being dried. To avoid this mistake, the experimenter

was careful when drying the surface of the eggs. A major problem that occurred

was lack of time. The eggs were not able to sit for the whole 72 hours that was

originally planned. There were also some calculation errors. Due to lack of time,

when the eggs were getting their masses taken time was running short.

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Acknowledgements

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Materials:

35 Eggs Needle

Procedures:

1. Grip 1 egg firmly in one hand and the needle in the other.

2. Apply pressure to the thin side of the egg until a hole appears.

3. Turn the egg around and apply pressure until a hole appears again.

5. Put your mouth on the small hole while blowing until all the egg falls out.

7. Rinse each eggs thoroughly, removing as much yolk from the shell as

possible.

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The equation to calculate the molarity of the acid solutions the grams to moles

grams per liter of citric acid found in orange juice meaning that is the

The figure below shows the conversion of 0.6 molarity citric acid to 0.047 molarity

Figure 16. Calculation of 0.6 molarity citric acid to 0.47 molarity citric acid.

The figure above shows the math to calculate the solution of citric acid from a 0.6

molarity to 0.47. According to the math, for every 66.6 mL of 0.6 molarity of citric

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The math above shows the calculation to find the amount of 14.8 molarity

phosphoric acid to add to form 1 liter of 0.057 molarity phosphoric acid which

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Appendix C: Randomization

Materials:

Ti-nspire calculator

Procedure:

2. Hit the menu button and scroll to “5: Probability” and press enter.

one comma three.

9. If the calculator shows a 1, it goes into the citric group. If its a 2, its an egg

in the phosphoric group. If it shows a 3, it will be in the water.

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To calculate the t value, following equations were used. The letter “t” equals the

mean of the first list, x̄1, subtracted by the mean of the second list, x̄2, all over

the square root of the sample deviation of the first list, s1 squared over the

number of trials for the list, n1, added together with the sample deviation of the

second list, s2, squared over the number of trials for the second list, n2.

Figure 18 shows the basic equation for a Two Sample t-Test with no

values entered.

Figure 19 shows the values plugged into the formula for a Two Sample t-

Test. After calculating this the t value was found to be -4.97536. Using Table C,

the p value was determined. The p value is the probability of obtaining results

that are equal to or more extreme than what was actually observed when the null

hypothesis is true. The p value of this test was determined to be 0.000016. The

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fact that the p-value is so much lower than the alpha level provides strong

Figure 20 includes the data that is substituted into the formula for a Two

Sample t-Test. After the formula was run the t value was determined to be

approximately -2.29514. This t value was then used to on Table C in order to find

0.017947. Due to this value being below the alpha level of 0.05 it is evidence.

Figure 20. Substitution for Two Sample t-Test of Water and Citric Acid

Figure 20 shows the formula that was used to get the t value of the Two

Sample t-Test. After performing the calculations it was found to be that the t

value was -8.29747. After finding the t value then p-value was found using Table

C. After using Table C the p-value appeared to be around 2.46878 x 10-7. Due to

the fact that the p-value is so low this is strong evidence against the null

hypothesis.

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<http://www.livestrong.com/article/147503-the-effect-of-citric-acid-on-

tooth-enamel/>

Cheng, Ran, Hui Yang, Mei-ying Shao, Tao Hu, and Xue-dong Zhou. "Dental

Erosion and Severe Tooth Decay Related to Soft Drinks: A Case Report

and Literature Review."

Journal of Zhejiang University. Science. B. Zhejiang University Press, May

2009. Web. 14 Mar. 2017.

<https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2676420/>.

<https://shwebook.com/search?q=tooth>.

Ehlen, Leslie A., Teresa A. Marshall, Fang Qian, James S. Wefel, and John J.

Warren. "Acidic Beverages Increase the Risk of in Vitro Tooth Erosion."

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<https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2516950/>.

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enamel.html>.

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Overview]."

Welcome to Karen's Science Communication Portfolio. Live Journal, 16

Feb. 2016. Web. 18 Mar. 2017.

<http://kquinto.livejournal.com/1794.html>

Re: How Are Teeth Similar to Eggshells? MadSci Netword, 20 May 1997.

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<http://www.madsci.org/posts/archives/1997-05/864330165.Ot.r.html>

Extracted Premolar Teeth."

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