Vous êtes sur la page 1sur 48

The Effect of Artificial Sweeteners on the Rate of Carbon Dioxide Produced during the

Fermentation of Yeast

Brianne Cochill, Krystal Quinn, and Aubrey Rupert

Macomb Mathematics Science Technology Center

10B

Mrs. Dewey / Mrs. Hilliard / Mr. Supal

24 May, 2017
Cochill - Quinn - Rupert 2

Table of Contents

Introduction………………………………………………………………………………....3

Review of Literature………………………………………………………………………...5

Problem Statement…………………………………………………………………………..13

Experimental Design………………………………………………………………………...15

Data and Observations………………………………………………………………………17

Data Analysis and Interpretation…………………………………………………………….27

Conclusion…………………………………………………………………………………...37

Appendix A Logger Pro and Gas Pressure Sensor Setup…………………………………....41

Appendix B Sample Calculations…………………………………………………………....42

Works Cited………………………………………………………………………………….46
Cochill - Quinn - Rupert 3

Introduction

Artificial sweeteners are a significant part of people's diet all across the world.

The sweeteners are included in everything from pop to baked goods. There has been an increase

in the popularity of artificial sweeteners because in the United States 9.3% of the population has

been diagnosed with diabetes. Many of these people have trouble finding suitable foods to eat,

because they cannot consume large amounts of glucose. One common food that diabetics cannot

eat is bread. This is because of the amount of sugar it requires to produce it. Alternative

sweeteners, such as Splenda and Stevia could be used in the bread making process to make it

more suitable for diabetics.

The experiment was designed to test if artificial sweeteners could carry out the

fermentation process of yeast as efficiently as glucose. If they are able to carry out this process,

they can be used in bread as an effective alternative to glucose. This was done by recording the

amount of byproduct, carbon dioxide, produced. Three test tubes were placed in a water bath

with a temperature between thirty-five and forty degrees Celsius. A yeast solution was then

placed into the test tubes followed by the sweetener solutions. All three of the test tubes were

then corked with a rubber stopper to contain and measure the carbon dioxide that was being

produced. To measure the carbon dioxide, the pressure within the test tubes during the reaction

was recorded using a gas pressure sensor and a logger pro.

It was hypothesized that glucose would yield the greatest changes in pressure followed by

Stevia and Splenda. It was also hypothesized that if Splenda and Stevia were compared to

glucose individually by running two-sample t-tests, that Splenda and Stevia would both have as

much as, if not more of an effect on the carbon dioxide production than glucose. These outcomes
Cochill - Quinn - Rupert 4

were hypothesized due to the molecular structure of each sweetener. Yeast uses glucose as its

main food source, and the glucose enables the yeast molecule to reproduce and then ferment. The

data collected from the experiment proved both of the hypotheses to be correct. The hypotheses

are also backed by other experiments done by researchers within the scientific field of

biochemistry. Although it is backed by previous research, glucose still performed the best and is

suggested to be used while baking to produce the best quality. Other factors of Splenda and

Stevia such as stability in heat also have effects on whether or not it can be used in baking.

Overall, the experiment was a success in proving that alternative sugars are capable of

fermenting yeast.
Cochill - Quinn - Rupert 5

Review of Literature

Sugar increases the rate at which yeast grows. The reproduction rate is affected by the

different types of sugar that are used. Granulated sugar, Splenda, and Splenda are suspected to

each cause a different reproduction rate of yeast. Approximately 1.25 million Americans have

been diagnosed with type one diabetes in the United States. ​Type one diabetes occurs when the

body lacks the ability to produce insulin​. The pancreas doesn't secrete insulin which causes a

buildup of glucose in the bloodstream. Glucose is the molecule that granulated sugar and natural

sugars are composed of. Artificial sweeteners are a lower dose of glucose making them a better

alternative. The purpose of this experiment is to determine if artificial sweeteners can be used in

bread as a substitute for other sugars. This will be done by measuring the amount of CO​2​ that is

produced, in kPa, or kilopascals, during the fermentation process with each sugar. ​To better

understand the topic, it is important to know how these sugars and yeast work.

This experiment will be using active dry yeast, so that when it is exposed to water it will

start to activate. Yeast is a single celled fungi that uses sugar to create energy. This process is

more efficient when oxygen is present, however if oxygen is not present, fermentation will occur

in place of respiration. When bread is created, the yeast cells undergo the fermentation process

because there is little to no oxygen present.

As glucose is added to the yeast, the fermentation process begins. Yeast derives

energy from simple sugars and complex starches. Glucose is the yeast’s main food source. All of

the artificial sweeteners chosen to be tested are complex sugars, but ​yeast can use its own

enzymes to break down more complex molecules into simple glucose ones. When the glucose

molecules are being broken down, the CO​2​ is released. ​The yeast will undergo fermentation
Cochill - Quinn - Rupert 6

which creates ATP, and produces carbon dioxide and ethanol as a byproduct. ATP is the energy

that the cell uses, and the carbon dioxide is what allows the bread to rise when it is baked with

yeast. This is because the carbon dioxide rises in the bread, creating air bubbles which is what

gives bread the spongy texture, in turn making the bread rise.

The first process that occurs during fermentation is called glycolysis. Glycolysis is an

anaerobic process, which means it will occur inside of a yeast cell regardless of if there is oxygen

present or not. During the process of glycolysis, start with a glucose molecule, and add two ATP

molecules. The addition and use of ATP also creates two molecules of ADP, which is used later

in this process. This ATP rearranges the glucose molecule and adds two phosphate atoms to the

molecule, creating a type of fructose. This fructose molecule is very unstable, so it breaks up into

a carbon three sugar molecule, and a DHAP molecule, which is also converted into the carbon

three sugar molecule. Next, this molecule of sugar goes through different reactions to create the

final product in the glycolysis cycle. A NAD+ is added, and this molecule turns into NADH.

This reaction will release energy, which allows for another phosphate to bond with the sugar

molecule. This new molecule then gives one of its phosphates to an ADP from the first half of

the cycle, which is above the dotted line in the diagram shown in Figure 1. This creates another

ATP molecule, and again changes the sugar molecule. This molecule is then converted again,

into an unstable molecule. This unstable molecule then adds a phosphate to a second group of

ADP that was created prior, which turns into another ATP molecule. The loss of this phosphate

creates pyruvate, which is the product of glycolysis, and will then be used in the alcoholic

fermentation process. It is important to note that two pyruvate molecules are created. This
Cochill - Quinn - Rupert 7

process occurs twice because two of the three carbon sugar molecules are formed, which is very

important, because it sets up the process for the carbon dioxide being created.

Figure 1. The Glycolysis cycle

The figure above is a drawing of the glycolysis cycle, which helps to better understand

where all of the different molecules are being added, as well as what is being produced.

After the glycolysis cycle, ether the respiration process would start, or the fermentation

process. Because of the lack of oxygen, for yeast the fermentation process will continue. There

are two different forms of fermentation, ether the alcohol or lactic acid processes. Yeast is an

alcohol fermenter, which means it produces NAD+, as well as ethanol and carbon dioxide as

byproducts (“Fermentation”). After the pyruvate molecules are created during glycolysis, they

release two carbon dioxide molecules that are byproducts, and are left with two molecules called

acetaldehyde, which are two carbon molecules. Then to transfer the NADH into NAD+, the

NADH that was created during glycolysis passes some electrons to the acetaldehyde, creating
Cochill - Quinn - Rupert 8

two NAD+ molecules which are used as energy, and ethanol which is also a byproduct (Khan

Academy).

Figure 2. The Fermentation Cycle

The figure above shows a model of the alcoholic fermentation process, and shows where

glycolysis converts into the regeneration process. This diagram is used to make the process

easier to follow, and shows exactly where the carbon dioxide will be released. This carbon

dioxide is extremely important because it is what will be measured during this experiment.

The experiment's purpose is to test to see if the yeast will be able to consume the artificial

sugars. In the experiment, the different types of sugars will be measured out in grams. Yeast

feeds off of sugar as explained above. Artificial sugars have a similar flavor to sugar but not the

exact same chemical makeup as shown below.


Cochill - Quinn - Rupert 9

Figure ​3.​ Stevia

Stevia is a sugar that is derived from a plant. Stevia is made up of three glucose

molecules and a diterpene. A diterpene is a molecule commonly found in plants and holds the

three glucose molecules together.

Figure 4. Splenda

Figure 4 above shows the artificial sweetener commonly known as Splenda. Its chemical

name is sucralose and its chemical formula is C​12​H​19​CO​8​. It is made up of chlorinated sucrose,

sucrose is a glucose molecule linked to a fructose molecule. The only difference between sucrose

and sucralose is that two hydroxides in the fructose molecule are swapped for chlorine

molecules.
Cochill - Quinn - Rupert 10

yes

Figure 5. Glucose

The figure above shows the molecule glucose which is located in natural sugars. Glucose

has a chemical formula of ​C​6​H​12​O​6​.This means that it is composed of six carbon atoms, twelve

hydrogen atoms, and six oxygen atoms. These were all produced through a natural process called

photosynthesis. Photosynthesis is a process in which sunlight is used to synthesize foods such as

glucose from CO​2​ and water.

The two artificial sugars and glucose all have very similar properties. All three contain

oxygen, which makes them somewhat similar in their chemical makeup. They also have similar

atomic structures, making them harder for the yeast to distinguish between the different types of

sugar.

Scientific studies have been done on the effect of artificial sugars on the reproduction rate

of yeast. A study was done by a group of high school students that measured the effect of

artificial sugars on the reproduction rate of yeast by measuring the amount of CO​2 ​that was

produced. With the help of their teacher, they came up with the conclusion that although

artificial sweeteners can be used as a substitute for natural sugars, it does not work as well. Less

CO​2​ was produced, meaning that the bread would not have risen as much as it would have if

natural sugars were used. Because this experiment was performed by high school students it may
Cochill - Quinn - Rupert 11

not be completely credible, but it supports the results of other prior research. A study done by

Tessa M​ ​Polakowski, a researcher at the Ohio Journal of Science, performed a similar

experiment. She compared the CO​2 ​production from the ​fermentation​ process between eleven

different types of sugars. The sugar with the highest CO​2​ production was sucrose, a natural

sugar. The natural sugars tended to have the highest amount of CO​2​. Although this is true, it was

concluded that the slight difference of CO​2​ production would not greatly affect the quality of the

bread.

There are a few lab procedures that have been chosen to follow during the course of the

experiment. A lab by “Science Buddies” used different types of sugars and compared the CO​2

production by using a homemade apparatus, which will not be used in this experiment. The idea

of using the different sugars and measuring the production of CO​2​ came from this source. It also

gave the temperature of ​110°F–115°F​, which the water will be heated to, that will be used

throughout this experiment. Another experiment that was used to create a new and unique lab for

the purpose of this experiment is from Vernier Software and Technology. They tested the Effect

of Temperature on Fermentation. This provided the setup that is used in the procedure of the new

experiment. The setup that was used includes a water bath, an Erlenmeyer, a logger pro with a

pressure sensor, a cork, and a clamp to hold the Erlenmeyer flask down. These two experiments

were a great help to making the following experiment that was used to test the effect of different

types of sugars on the reproduction rate of CO​2​.


Cochill - Quinn - Rupert 12

Figure 6. Set Up to Measure CO​2​ Production

The figure above shows the setup that Vernier Software and Technology used to measure

the amount of CO​2​ that was produced. The setup they used is very similar to the one that was

used in this experiment.

Although previous experiments have been done before, none are exactly like what this

one is trying to achieve. The experiment studies the reproduction of yeast using artificial sugar,

the amount of CO​2 produced


​ shows how well the yeast can break down the artificial sugars and

use them as energy. The yeast and the sugars will be measured in grams, and the CO​2 will
​ be

measured using a logger pro, in ppm. This experiment is fermentation, similar to the one done by

the ​Ohio Journal of Science. Science Buddies also did fermentation. This experiment tests to see

if artificial sugars can be used as a substitute for natural sugars such as glucose in items such as

bread. This was done by comparing the amount of ​CO​2 ​that was produced when the yeast went

through the fermentation process.


Cochill - Quinn - Rupert 13

Problem Statement

Problem Statement:

During the process of this experiment it will be determined if yeast is able to use artificial

sugars to carry out the fermentation process, and if so which sweetener, Glucose, Splenda, or

Stevia, will produce the most carbon dioxide byproduct measured in kilopascals per minute

during this process.

Hypothesis:

It was hypothesized that glucose would yield the greatest changes in pressure in kPa

followed by Stevia and Splenda. It was also hypothesized that if Splenda and Stevia were

compared to glucose individually by running two-sample t-tests, that Splenda and Stevia would

both have as much as, if not more of an effect on the carbon dioxide production than glucose.

Data:

Data in this experiment will be dissected through a two sample t-test. The independent

variables are the different types of sweeteners that will be used (Glucose, Splenda, and Stevia),

each will be measured in grams. The dependent variable is the carbon dioxide that is produced

during the fermentation process. This will be measured by using a logger pro with a gas pressure

sensor measuring in kilopascals (kPa). A few variables will be kept as constants to keep the trials

uniform, time in minutes and water which will be added in milliliters (mL). The control of this

experiment is glucose which will be measured in grams. Glucose is the control so the artificial

sweeteners have something to be compared to when determining the results of the experiment.
Cochill - Quinn - Rupert 14

Experimental Design

Materials:
40 g Stevia (6) 40 mL Test Tubes
40 g Splenda 1500 mL Water
40 g Granulated Sugar 7 g Active Dry Yeast
(1) Logger Pro (1) Scale (0.00 g precision)
(3) Gas Pressure Sensor (1) Weight Boat
(2) Thermometers (0.0 precision) (1) Ring Stand
(4) 3 mL Pipet (3) Test Tube Clamps
(3) Rubber Stoppers (Size 2) (2) Hot Plates
(3)C (1)1000 mL Beaker
(1) Test Tube Rack (1) 500 mL Beaker
(2) Hot Mitts (1) 250 mL Beaker
(1) Roll of Tape (1) Sharpie
Procedure:

*Note: Everything in this experiment the may come into contact with yeast at any time must be

thoroughly cleaned, this includes all glassware, pipettes, thermometers, weight boats, and

rubber stoppers

*Note: Label everything with tape and a sharpie, DO NOT mix up any of the sugars or yeast, or

the glassware/test tubes/pipettes/weight boats as this may lead to a skewed experiment, it is also

suggested to label the gas pressure sensors and plug them into the Logger Pro the same way

every time

1. First fill up the 1000 mL beaker to 800 mL with water and place it on a hot plate, turn the
hot plate on high and stick the thermometer in the beaker

2. Next fill the 500 mL beaker to 400 mL with water and place it on the second hot plate,
turn the hot plate on a medium setting and stick a thermometer in the beaker

3. Wait for the water to heat up to a temperature between 35-40 degrees Celsius, check the
thermometer regularly because this will go quickly, and if it gets too hot add a small
amount of cold water but DO NOT fill above the 1000 mL mark

4. While waiting for the water to heat up mass out the sugars, mass out all 3 sugars to 2
grams. Place the sugars in their respective 100 mL beakers, which should be labeled with
the tape and sharpie.
Cochill - Quinn - Rupert 15

5. Also mass out the yeast to 7 grams and place it in the 250 mL beaker

6. Next set up the ring stand with three test tube clamps like shown in Figure 1 below

7. Once the water in the 500 mL beaker is between 35 and 40 degrees Celsius, using two hot
mitts, pour 40 mL into each 100 mL sugar beaker, also pour 100 mL into the 250 mL
yeast beaker, gently stir the solutions with a pipet, use a DIFFERENT pipet for each
solution

8. Refill the 500 mL beaker with water and place it back on the hot plate at a low setting, it
may be needed later in the experiment

9. Now take the 1000 mL beaker, at its correct temperature, and place it on the ring stand,
directly under the test tube clamps, leave the thermometer in the beaker

10. Ready the Logger Pro and Gas Pressure sensor, follow the instructions in Appendix A,
make sure everything is ready before moving on

11. Carefully slide a test tube into each clamp, a little under half of the test tube should be in
the water, carefully tighten the clamp so that the test tube does not slide around

12. Take the pipet used to stir the yeast and carefully add 2.5 mL of yeast solution to each
test tube, try to avoid getting the solution on the sides of the test tube

13. Next add 2.5 mL of each sugar solution to its corresponding test tube, randomize this
order for every trial, right after adding the sugar to the test tube firmly put in the rubber
stopper, once all three are firmly stoppered and connected to the logger pro, start the trial

14. While waiting for the trial to complete, record any observations in the observations table

15. Once the trial is complete, take the stoppers out of the test tubes and carefully remove
them from the clamps, for now place them right side up in the test tube rack

16. Check the temperature of the water bath, if it is below 35 degrees Celsius add some hot
water from the 500 mL beaker, be careful not to overfill the 1000 mL beaker, remember
that test tubes must go in the water, if there is not enough room for extra water pour some
out of the 1000 mL beaker and add the hot water, be careful not to make it too hot

17. Take the second set of test tubes and repeat steps 9-13

18. While waiting for the second trial to complete wash the dirty test tubes and place them
upside down in the test tube rack to dry
Cochill - Quinn - Rupert 16

19. Once trials are completed save the data on a flash drive, the instructions for this can also
be found in Appendix A

20. Dispose of the sugar solutions, the yeast solution, and empty out all beakers down the
drain. Thoroughly rinse all of the glassware, to ensure that nothing gets contaminated.

21. Look at the data on a computer with the Logger Pro software, identify at what time the
reaction started and the pressure in kPa, as well as the peak pressure for each sugar and
the time that it peaked. Record them in the data table

Diagram:

Figure 7. Logger Pro Setup

Figure 7 above shows the setup that was used to measure the amount of carbon dioxide

produced by the yeast. The beaker rested in a water bath, and a tube lead from it to the Logger

Pro to measure the pressure of the gas being produced in kilopascals (kPa). The test tubes were

held up by the test tube clamps, which were attached to the ring stand. The ring stand and clamps

were not labeled in the diagram above because they are not really visible. Note that all test tubes

should be labeled to make sure sugars are not mixed. The beaker was also labeled, which is

recommending when running more than one trial at a time.


Cochill - Quinn - Rupert 17

Data and Observations

This experiment is measuring the amount of carbon dioxide byproduct being produced by

the fermentation of yeast, using three different types of sugar (Glucose, Splenda, and Stevia).

This carbon dioxide is being measured in kilopascals per minute.

Table 1
Rate of Carbon Dioxide Production Using Splenda
Splenda

Trial Initial Final Initial Final Time Rate of


Pressure Pressure Time (minutes) Reproduction of
(kPa) (kPa) (minutes) Yeast
(kPa/minutes)

1 104.621 121.345 3.500 15.000 1.454

2 105.260 130.083 1.000 15.000 1.773

3 102.936 109.715 2.500 12.500 0.678

4 98.126 108.344 2.000 15.000 0.786

5 101.954 109.467 2.500 15.000 0.601

6 103.478 117.090 2.500 13.500 1.237

7 102.180 111.105 3.000 13.500 0.850

8 100.429 110.538 4.500 15.000 0.963

9 4.967 26.568 3.500 15.000 1.878

10 100.865 102.787 2.500 14.000 0.167

11 105.666 120.981 4.500 15.000 1.459

12 105.666 129.910 2.500 15.000 1.940

13 105.329 125.954 2.500 15.000 1.650

14 99.758 107.216 2.500 12.500 0.746


Cochill - Quinn - Rupert 18

Splenda

Trial Initial Final Initial Final Time Rate of


Pressure Pressure Time (minutes) Reproduction of
(kPa) (kPa) (minutes) Yeast
(kPa/minutes)

16 102.769 113.229 3.000 13.000 1.046

17 101.308 108.112 4.500 15.000 0.648

18 104.424 127.760 4.000 15.000 2.121

19 102.617 110.190 3.500 14.000 0.721

20 104.538 118.838 4.000 15.000 1.300

Average 1.138

The table above shows the results from the twenty trials that were conducted using

Splenda. The rate of reproduction of yeast was calculated in kilopascals per minute using the

data in the table. Twenty trials were conducted using Splenda.

Table 2
Rate of Carbon Dioxide Production Using Stevia
Stevia

Trial Initial Final Initial Final Time Rate of


Pressure Pressure Time (minutes) Reproduction of
(kPa) (kPa) (minutes) Yeast
(kPa/minutes)

1 103.781 116.419 3.500 15.000 1.099

2 105.721 124.306 2.500 15.000 1.487

3 103.799 132.175 2.500 15.000 2.270

4 100.061 102.823 3.000 11.500 0.325

5 101.806 124.802 0.500 15.000 1.586


Cochill - Quinn - Rupert 19

Stevia

Trial Initial Final Initial Final Time Rate of


Pressure Pressure Time (minutes) Reproduction of
(kPa) (kPa) (minutes) Yeast
(kPa/minutes)

7 102.543 117.738 3.500 15.000 1.321

8 98.813 117.288 3.000 15.000 1.540

9 100.970 109.951 4.000 15.000 0.816

10 102.213 114.354 2.500 15.000 0.971

11 104.121 122.934 3.000 15.000 1.568

12 105.476 127.395 3.000 15.000 1.827

13 104.969 124.458 2.500 12.500 1.949

14 101.849 130.628 2.500 15.000 2.302

15 102.249 125.101 2.000 15.000 1.758

16 103.033 124.469 2.000 15.000 1.649

17 101.795 113.724 3.000 15.000 0.994

18 100.277 102.143 3.000 13.000 0.187

19 104.891 120.053 3.500 15.000 1.318

20 103.579 120.567 3.500 15.000 1.477

Average 1.391

The rate of reproduction of yeast in kilopascals per minute was calculated using the data

from the table above. The pressure of the carbon dioxide in each trial was recorded along with

the time at which they occurred at. Twenty trials were completed in total using Stevia.
Cochill - Quinn - Rupert 20

Table 3
Rate of Carbon Dioxide Production Using Glucose
Glucose

Trial Initial Final Initial Final Time Rate of


Pressure Pressure Time (minutes) Reproduction of
(kPa) (kPa) (minutes) Yeast
(kPa/minutes)

1 103.476 122.352 2.000 15.000 1.452

2 104.575 127.505 2.500 15.000 1.834

3 101.784 115.912 2.000 12.000 1.413

4 102.330 121.051 0.000 15.000 1.248

5 103.948 125.246 2.000 15.000 1.638

6 103.838 130.271 2.000 15.000 2.033

7 102.933 127.561 1.500 15.000 1.824

8 99.486 120.554 3.000 15.000 1.756

9 101.437 120.941 2.500 15.000 1.560

10 106.582 132.359 2.000 15.000 1.983

11 107.172 128.536 2.500 15.000 1.709

12 106.551 131.080 2.500 15.000 1.962

13 103.252 122.469 3.500 15.000 1.671

14 103.295 125.741 2.000 15.000 1.727

15 103.749 122.012 1.500 14.000 1.461

16 102.873 126.168 1.500 15.000 1.726

17 102.423 118.795 2.500 15.000 1.310

18 103.795 123.455 3.000 15.000 1.638


Cochill - Quinn - Rupert 21

Glucose

Trial Initial Final Initial Final Time Rate of


Pressure Pressure Time (minutes) Reproduction of
(kPa) (kPa) (minutes) Yeast
(kPa/minutes)

20 103.359 120.320 3.500 15.000 1.475

Average 1.648

The goal of trials was to find the rate of reproduction of yeast in kilopascals per minute of

carbon dioxide being produced. This is shown Table 3 above, in the case of Glucose. Each

number that is needed for the final equation is shown in the table, and is used to find the rate of

reproduction. There were twenty trials completed using the glucose sugar.

Table 4
Observation Table for Splenda
Trials
Trial Splenda
1 Very few bubbles at the beginning. Water and bubbles start separating. Separates
the slowest. Made a popping sound when uncorked
2 Didn’t make a popping sound when uncorked
3 Warm water was added to keep temperatures at a constant level. Made a popping
sound when uncorked.
4 Super low pressure, almost unchanging, super low water bath temp, didn’t make a
popping sound when uncorked
5 Didn’t pop when uncorked
6 Made a popping sound when uncorked.
7 At 11 minutes, it has the lowest pressure, however there are more bubbles than the
rest. At 15 minutes the pressure didn’t change. Made a popping sound when
uncorked
8 Made a popping sound when uncorked
Cochill - Quinn - Rupert 22

Trials

Trial
Splenda
10 Hot water was added, increased temp a few degrees. Made a popping sound when
uncorked
11 Added some hot water. It had most bubbles. Made a popping sound when uncorked.
12 Water slightly cooler. It spiked about 3 min in. The cork popped when taken off
13 The graph spiked. Made a popping sound when uncorked.
14 Did not make a popping sound when uncorked
15 Pressure extremely low, didn’t make a popping noise
16 Did not make a popping sound when uncorked
17 Splenda flat line (lowest), might be something with sensor, didn’t make a popping
sound when uncorked
18 Hot water was added, increased temp a few degrees, didn’t make a popping sound
when uncorked
19 Added hot water to water bath. Spiked at 5 minutes, it had the most bubbles, didn’t
make a popping sound when uncorked
20 Made a popping sound when uncorked

Table 4 above shows the observations made on each trial that used Splenda. The data for

all twenty trials was recorded. A few trials did not go as planned, and some went exceptionally

well. All of this can be found in the above table.


Cochill - Quinn - Rupert 23

Table 5
Observation Table for Stevia
Trials
Trial Stevia
1 Larger bubbles at the beginning. Water and bubbles start separating
2 When the cork was taken off of the test tube at the end of the trial, a popping noise
was heard
3 Bubbles have more air, when the cork was taken off of the test tube at the end of the
trial, a popping noise was heard
4 Super low water bath temp, when the cork was taken off of the test tube at the end
of the trial, a popping noise was not heard
5 Creates a huge air bubble, when the cork was taken off of the test tube at the end of
the trial, a popping noise was heard
6 At 11 minutes stevia has the most amount of bubbles, when the cork was taken off
of the test tube at the end of the trial, a popping noise was heard
7 When the cork was taken off of the test tube at the end of the trial, a popping noise
was heard
8 There is a huge clear bubble sitting on top of the solution, when the cork was taken
off of the test tube at the end of the trial, a popping noise was heard
9 Graph spiked at 5 minutes. When the cork was taken off of the test tube at the end
of the trial, a popping noise was heard
10 Stopper popped off by itself at 13.5 min, was replaced to complete trial, when the
cork was taken off of the test tube at the end of the trial, a popping noise was heard
11 When the cork was taken off of the test tube at the end of the trial, a popping noise
was heard
12 Spiked about 3 min in, when the cork was taken off of the test tube at the end of the
trial, a popping noise was heard
13 The graph spiked, when the cork was taken off of the test tube at the end of the trial,
a popping noise was heard
14 When the cork was taken off of the test tube at the end of the trial, a popping noise
was not heard
15 When the cork was taken off of the test tube at the end of the trial, a popping noise
was heard
Cochill - Quinn - Rupert 24

Trials

Trial Stevia

17 A test tube broke before start, had to be replaced, should not have affectd data,
when the cork was taken off of the test tube at the end of the trial, a popping noise
was not heard
18 very curvy graph, when the cork was taken off of the test tube at the end of the trial,
a popping noise was not heard
19 When the cork was taken off of the test tube at the end of the trial, a popping noise
was not heard
20 Highest pressure out of the three with 3 minutes left, when the cork was taken off of
the test tube at the end of the trial, a popping noise was heard

Table 5 above and on the previous page shows the observations that were made during

each trial that included Stevia. There was a total of twenty trials. All of the flaws in the

procedure, random outliers, etc. were mentioned.

Table 6
Observation Table for Glucose
Trials
Trial Glucose
1 Bubbles forming rapidly inside of test tubes. The water at bubbles separated, at a
quick rate and are cream colored. The glucose rate spiked
2 At the eleven-minute mark glucose had the highest pressure, and when the cork was
taken off of the test tube at the end of the trial, a popping noise was heard
3 At ten minutes, there was no separation between bubbles and water. At the end there
is still no separation
4 Super low water bath temp, no pop There was a very low water bath temperature
when the trial started. when the cork was taken off of the test tube at the end of the
trial, a popping noise was not heard
5 The pressure is super high, it is suspected to be because of the high temperature of
the water bath. when the cork was taken off of the test tube at the end of the trial, a
popping noise was heard
Cochill - Quinn - Rupert 25

Trials
Trial Glucose
7 When the cork was taken off of the test tube at the end of the trial, a popping noise
was heard
8 When the cork was taken off of the test tube at the end of the trial, a popping noise
was heard
9 The graph spiked at 5 minutes when the cork was taken off of the test tube at the
end of the trial, a popping noise was heard
10 Glucose had the lowest graph, when the cork was taken off of the test tube at the
end of the trial, a popping noise was not heard
11
All very close together ie little diff in pressure, also consistent upward trend, popped
12 The graph spiked about 3 min in, when the cork was taken off of the test tube at the
end of the trial, a popping noise was heard
13 The graph stopped increasing. When the cork was taken off of the test tube at the
end of the trial, a popping noise was heard
14 Has the most bubbles out of the three test tubes at 15 minutes, when the cork was
taken off of the test tube at the end of the trial, a popping noise was not heard
15 Bubbling a lot. It looks like the bottom is filled with water, or something clear with
yeast bubbles on the top, when the cork was taken off of the test tube at the end of
the trial, a popping noise was heard
16 When the cork was taken off of the test tube at the end of the trial, a popping noise
was not heard
17 Glucose has the most bubbles, when the cork was taken off of the test tube at the
end of the trial, a popping noise was heard
18 Very curvy graph, when the cork was taken off of the test tube at the end of the trial,
a popping noise was heard
Cochill - Quinn - Rupert 26

Trials
Trial Glucose
19 Glucose dropped at 5 minutes, when the cork was taken off of the test tube at the
end of the trial, a popping noise was heard
20 When the cork was taken off of the test tube at the end of the trial, a popping noise
was heard

The table above and on the previous page show the observations that were made during

the trials that involved glucose. Table 6 shows all twenty trials and all of the occurrences within

them.
Cochill - Quinn - Rupert 27

Data Analysis and Interpretation

Yeast is a living organism that needs sugar and water to reproduce. During its

reproduction, carbon dioxide is produced. This research experiment tested the effect of different

sweeteners on the rate of fermentation of yeast. The rate was calculated in kilopascals per minute

by measuring the pressure of the carbon dioxide in kilopascals within the test tubes over a period

of fifteen minutes. Using a Logger Pro and a gas pressure sensor, this data was able to be

collected. The control of the experiment was the glucose because it is the sweetener, and natural

sugar that is used in almost all products that need yeast and is proven to work. Other similar

experiments have been conducted using different types of sweeteners. These results were similar

to the ones that were received during this research experiment. The order in which trials were

conducted was randomized by using a calculator. Splenda was number one, Stevia was number

two, and glucose was number three. The randint(); function was used and the order in which

these numbers appeared in was the order the trials were conducted in. Two-sample T-tests were

used to analyze the data. T-tests were used so the artificial sweetener could be compared with

the glucose, which would be acting as a standard. These were conducted to compare the Splenda

and Stevia to the results of the glucose in order to see which performed the best.

Although the experiment has been found to be reliable through repetition and

randomization, it also must be considered “normal” in order for a two sample t-test to be

conducted. Two two-sample t-tests were performed in order to compare the results of Splenda to

Glucose and the results of Stevia to Glucose. The box plots below show each of the data sets

separately and then all on one plane for comparison. They also show if any outliers, or skewness
Cochill - Quinn - Rupert 28

is present. Box plots are a good visual representation of the data, because the key points are

identified to clarify where the data occurs.

Figure 8. Splenda Box Plot

The figure above shows the data collected for Splenda plotted in a box plot. The median

is 1.0045, meaning that fifty percent of the data occurs before the point 1.0045. This point is

close to the middle, but slightly closer to the Q1 value, so it can be concluded that the data was

only slightly right skewed. The slight skewness to the right is not enough to be considered

abnormal. The data also has no outliers, further enforcing that it is a normally distributed data

set. The data set also has a mean, or average of 1.138. When comparing this to the median of the

data, it can be inferred that the data had a large amount of variability which is why these two

numbers were so far apart. This box plot also shows the range of the data, which is 1.945. This is

a fairly large range that shows that there is a lot of variability in the data, this may point to

possible flaws in the experiment.


Cochill - Quinn - Rupert 29

Figure 9. Stevia Box Plot

The figure above shows the data collected for Stevia plotted in a box plot. The median is

1.482. This means that fifty percent of the data is before the point 1.482 and the other fifty

percent is after, which is slightly higher than Splenda. There is a slight skewness to the left, but

nothing major enough to be considered abnormal. The skewness of the graph could also be

because of the difference between the median and mean values. The mean, or average of the

Stevia data set was 1.391. Since these two are closer together, the graph has less variability,

however it does look like there was almost an outlier on the left side, causing the left skewness.

Because the data above has no outliers it is considered a normal distribution, allowing it to be

used in a t-test. The range is 2.115, this is an even larger range than Splenda, showing more

variability. This variability may later point to faults in the experiment or in human error.
Cochill - Quinn - Rupert 30

Figure 10. Glucose Box Plot

The figure above shows the data collected for glucose plotted in a box plot. The median

is 1.6735. Fifty percent of the data comes before this point and fifty percent of it comes after.

Glucose has the largest median out of all three sweeteners. There is a slight skewness to the left,

but not enough to be considered abnormal. This skewness is small compared to the other

sweeteners tested, and this is because of the median and mean. The mean of this data set, or

1.648 is very close to the median. This implies that there wasn't a lot of variability, which makes

sense considering these trails were treated as standards.

The absence of outliers enforces that box plot above is a normal distribution, which allows the

data to be tested in t-test. The range is 0.785, this is the smallest range out of all three

sweeteners, showing the least amount of variability. This is normal, considering that glucose was

used as the control during the experiment.


Cochill - Quinn - Rupert 31

Figure 11. Comparing Box Plots

The figure above shows all three sugars graphed next to each other. Figure 11 above

shows that the three sweeteners are within the same vicinity, but each have their differences. The

median of glucose was the highest, the Stevia was in the middle, and Splenda had the lowest.

However, glucose had the least amount of variability, Splenda had a larger range, and Stevia had

the largest variability. This means that the data could be slightly skewed, whether it was because

of the stoppers in the test tubes, faulty lab quests, or the sensitivity of the yeast organisms. The

box plots also show that all of the data overlaps at some point. For example, the third and fourth

quartile of Splenda overlap second, third, and fourth quartile of Stevia. Where they overlap is

where they shared similar data. The box plots provide a lot of information on how the trials

within the experiment ran.

This experiment was analyzed by using two sample t-tests. One two-sample t-test was

conducted to compare Splenda to glucose, a second one was conducted to compare Stevia to

glucose. Glucose was used as the control due to its chemical properties and its proven
Cochill - Quinn - Rupert 32

effectiveness. Two-sample t-tests were used to compare the two sweeteners to glucose. There

were no outliers in any of the data meaning that it was appropriate to use this test.

H​o :​ μ g = μ s H​o :​ μ g = μ t

H​a ​: μ g ≤ μ t H​a ​: μ g ≤ μ t

Figure 12. Statistical Hypothesis

The hypothesis used for these two-sample t-tests are stated in the figure above. Each μ

symbol represents the mean, and is differentiated from each different variable marked as a

subscript next to it, t standing for Stevia, s for Splenda, and g for glucose. The first two on the

top are the null hypothesis, or ​H​o​ . There is a null hypothesis for each t-test being performed, one

for Splenda and one for Stevia. They are saying that the mean pressure production rate of

glucose will be equal to the mean pressure production rate of Splenda or

Stevia. The bottom equations are the alternative hypothesis, which are represented by ​H​a​. ​The

alternative ​hypothesis states that the mean pressure production of glucose will be less than or

equal to the mean pressure production rate of Splenda or Stevia.

During any statistical analysis a few assumptions have to be made. The first assumption

is that the test was a simple random sample meaning that there was little to no bias in gathering

the data. This can be proven by the box plots in Figure 11. The second assumption is that the

total population is greater than or equal to ten times the sample population, which is true in this

experiment because twenty trials were conducted and twenty times ten is two hundred and places

such as bakeries use these sweeteners every day, making the population much larger. The last

assumption is that the sample is large, meaning it is greater than thirty. In this experiment, the
Cochill - Quinn - Rupert 33

sample is not large, so the box plots in Figure 8, 9, 10, and 11, were used to determine if the

results could be considered normal, and they all were.

The t-values for comparing Stevia to Glucose and comparing Splenda to Glucose were

calculated using the equation listed in Figure B2 in Appendix B. The p-graph is a visual

representation of the significance, also known as the efficiency of producing carbon dioxide

during the fermentation process of yeast, of Splenda and Stevia compared to Glucose.

Table 7
Standard Deviation Values
Standard Deviation

Glucose Splenda Stevia

0.219 0.537 0.551

The table above shows the standard deviations of Glucose, Splenda, and Stevia. Glucose

had the lowest standard deviation, Splenda had the second highest, and Stevia had the highest

standard deviation.
Cochill - Quinn - Rupert 34

t = 3.9348

Figure 13. T-Test comparing Splenda and Glucose

The figure above shows all of the previously found variables plugged into the two

sample t-test equation, found in appendix b, to compare glucose and Splenda. Using the degree

of freedom 19 (n-1), and the C Table, the p-value is less than 0.0005. This means that it is

statistically significant at the alpha level 0.05, and even at the alpha level 0.01. This alpha level

would be shown in the last 99% of a p-graph, in this case in the red box shown above, where the

area beneath the bell curve is not shaded. Note that the p value is not shaded because this graph

is not looking for the tail probability, which is what needs to be found in order to synthesis the

data.

While comparing Splenda and glucose, the null hypothesis, ​H​o ​: μ g = μ s would be

rejected. The hypothesis is rejected because the p-value is less than the alpha level of 0.05. There

is evidence that supports yeast can produce the same, or higher amounts of carbon dioxide when

reacting with Splenda instead of glucose. There is less than a 0.05% chance that the data could

have been obtained by chance alone if the null hypothesis was true.
Cochill - Quinn - Rupert 35

t = 1.9426

Figure 14. T-Test comparing Stevia and Glucose

The figure above shows all of the previously found variables plugged into the two

sample t-test equation, found in appendix b, to compare glucose and stevia. Using the degree of

freedom 19 (n-1), and the C Table, the p-value is between 0.025-0.05, this means that it is

statistically significant at the alpha level of 0.05. This value is shown in the p-graph above, and

would be found somewhere in the red box where the graph is not shaded. In the table shown, the

p value is different. This is because the p value given has to be subtracted from 1 because the p

value is looking for the tail probability. When done on a calculator, this is not done which is why

the part where it is not shaded is the correct p value.

When comparing stevia and glucose, the null hypothesis, ​H​o :​ μ g = μ t in this case would

be rejected. This is because the p-value is less than the alpha level of 0.05. There is evidence to

support that yeast produces more or the same amount of carbon dioxide while reacting with

Stevia, than with regular glucose. There is between a 2.5% and 5% chance that this data could

have been obtained by chance alone if the null hypothesis was true.
Cochill - Quinn - Rupert 36

Because of the two two-sample t-tests conducted, it can be concluded that Splenda and

Stevia will work just as well, or even better when reacting with yeast to create carbon dioxide.

This is shown in both two-sample t-tests that were run in Figures 13 and 14 above. Both of the

null hypothesis were rejected, meaning that the p-value was lower than the alpha level. There is

evidence to support the alternative hypothesis that stated that Splenda and Stevia would produce

just as much carbon dioxide if not more during the reproduction process of yeast. This data is

also significant because between both of the tests, there was a maximum of a 5% chance that the

data could be created by chance alone, as long as the null hypothesis is true. This means that it

would be extremely hard to get the data by chance alone, which makes the data collected more

valid. The two-sample t-tests show strong evidence that Splenda would be able to work as a

substitute for glucose in the reproduction process of yeast. They also show that Stevia would

most likely work as a glucose substitute in yeast, however Splenda would work better.
Cochill - Quinn - Rupert 37

Conclusion

The objective of this experiment was to determine if yeast was able to use artificial

sweeteners to carry out the fermentation process. Since carbon dioxide is a byproduct of

fermentation, the pressure changes that occurred due to its production, were used to quantify

fermentation for this experiment.

The hypothesis that glucose would yield the greatest changes in pressure followed by

Stevia and Splenda was accepted. It was also hypothesized that if Splenda and Stevia were

compared to glucose individually by running two-sample t-tests, that Splenda and Stevia would

both have as much or more of an effect on the carbon dioxide production than glucose. This

hypothesis was also accepted. It was determined that glucose produced the highest median

change in pressure. Glucose produced the median rate of 1.6735, means that every minute, the

pressure of the carbon dioxide within the test tube went up by approximately 1.6735 kPa.

Splenda produced a median rate of 1.0045 and Stevia produced a median rate of 1.482,

which means that both alternative sweeteners are capable of carrying out the fermentation

process of yeast. This data was used in the two-sample t-tests. Splenda produced the t-value of

3.9348 and the p-value of less than 0.0005, and Stevia produced the t-value of 1.9426 and a

p-value between 0.025-0.05. This means that both alternative sweeteners worked just as well as

if not better than glucose at producing carbon dioxide from fermentation. All three sweeteners

can be used as effective ways to carry out the fermentation process of yeast. The results agree

with the previous research that concluded that the respective alternative sweeteners were found

to be viable substitutes for glucose,​ ​however glucose still produced the most carbon dioxide

pressure during the fermentation process of yeast, and is recommended to yield the best results.
Cochill - Quinn - Rupert 38

Stevia and Splenda both function in the fermentation process of yeast because their

molecular structures contain glucose molecules. Glucose is the main source of food for yeast,

which is why the fermentation process can occur. However, Splenda and Stevia do not perform

as well because they have other molecules within them, which the yeast is not able to break

down. In Splenda there is a chlorinated fructose molecule bonded to the glucose molecule, and in

Stevia a diterpene molecule bonded to the three glucose molecules. Stevia has a greater change

in pressure than Splenda because it has three glucose molecules compared to the single glucose

molecule in Splenda (Jaksich). Also, Stevia is all natural, making it slightly easier to break down

than Splenda. Since glucose is the sugar that yeast naturally consumes and is the easiest of the

three to break down, explaining why it has the greatest change in pressure.

While the experimental design was efficient, the lack of time led to errors that could have

been improved. For example, when doing multiple trials in a row, the water bath temperatures

would decrease, and were hard to keep at a consistent temperature which could have led to faulty

data. This could have caused the yeast to start to die since the yeast needs to be kept between

thirty five and forty degrees Celsius. In future research, this could be avoided by using a larger

water bath in which more trials can be run at a time with the same conditions. The temperature

could be kept constant by using a fish tank heater or heating pads. Other small details were

glossed over such as how well the stoppers were fit into each individual test tube which could

have affected the pressure inside the tube, causing a fault in any data. Also, the height that the

test tubes were at varied and determined how much of the yeast was kept in the water bath, if not

all of the yeast was fully submerged, it could have died, leading to faulty data.
Cochill - Quinn - Rupert 39

Another error was that, while buying the packets of Splenda and Stevia, the containers

read that each packet contained two grams. This was interpreted as two grams of sugar in each

package, but the two grams referred to the volume of the packets instead of the mass of the

sugar. This was not realized until halfway through the experiment, so the same packets were

used in order to keep the results consistent. If this could be redone, it is suggested to mass each

sugar in every trial instead of using the packages. Also, the gas pressure sensors used did not

always work properly. After the trials were started, it was determined that the gas pressure sensor

did not work if the pressure was not moving at all but bubbles were forming, indicating that the

pressure should have gone up because of the increase of carbon dioxide. To fix this, check the

gas pressure sensors before trials are conducted, or consider using a method such as an inverted

test tube submerged in a water bath.

To expand the research, the structure of the sugars could be compared to glucose more in

depth to see if similar structures produce higher rates of carbon dioxide production. Also, it

could be determined if these results would still occur in a more practical sense such as in a

bakery by baking something such as bread, one loaf with glucose, one with Splenda, and one

with Stevia, and comparing the quality. Possible further research is changing the temperature of

the solutions and water during the trials to see the ideal temperature for this process. Also, using

a combination of sugars to see if they would interact could be tested, or even testing more types

of sweeteners such as honey to see how the yeast would react to them. Glucose, Splenda, and

Stevia are all able to carry out the fermentation process of yeast and can not only be used in the

science lab, but in the real world. Industries such as bakeries and bread shops could benefit from

the information in this experiment by using Glucose for the cheapest and best quality product, or
Cochill - Quinn - Rupert 40

Splenda and Stevia as a backup or an alternative for diabetics. Diabetics could also benefit from

knowing that Splenda and Stevia are able to be used as a substitute for glucose due to the fact

that diabetics cannot have glucose. Diabetics cannot have an abundance of glucose because their

bodies do not know how to produce enough insulin to break it down quickly enough, so the use

of Splenda and Stevia would lessen their total intake of Glucose.


Cochill - Quinn - Rupert 41

Appendix A: How to Setup the Logger Pro and Gas Pressure Sensor

1. After turning on the Logger Pro, plug in all three gas pressure sensors at the top of the
device.

2. The gas pressure sensors should have thin drills connected to the tubing and they should
then be securely screwed into rubber stoppers.

3. On the home screen three boxes should show the current pressure that is being measured
by the three sensors, if the measurements are not measuring gas pressure in kilopascals
either unplug the sensor and then re-plug it in, or manually set it to gas pressure sensor in
kilopascals in the settings.

4. Click Experiment then data collection. Change the collection time to 15 minutes, also
ensure that data is being collected every thirty seconds.

5. Once the experiment is ready press start and wait for the data collection to complete.

6. After data collection is complete, stop the trial and save the data.
Cochill - Quinn - Rupert 42

Appendix B: Sample Calculations

The following​ ​figures are sample calculations used to analyze and interpret the data that

was collected. The rate of carbon dioxide production, t value, and standard deviation calculations

are shown.

To analyze the data, the following equation where the final pressure minus the initial pressure

was divided by the final time minus the initial time.


y2−y1 f inal pressure(kP a) − initial pressure(kP a)
x2−x1
= f inal time(min)− initial time(min)

Figure B1 below shows a sample calculation to find the rate of carbon dioxide production.
y2−y1
Rate = x2−x1

= 121.345−104.621
15.000−3.500

= 1.454 kPa/minute

Figure B1. Rate of Carbon Dioxide Production Equation

Sample Calculation of the rate of carbon dioxide production using the first trial of the data. The

initial pressure and initial time were decided based on when the reaction occurred. It took

between one and four minutes for this to occur. It was determined by looking at the graph and

taking the first point in which showed signs of linear relations to the other points. The final

pressure and time were decided by taking the point at which the reaction stopped and recording

the time and pressure it was at. This was determined by taking the point when the pressure

stopped increasing.
Cochill - Quinn - Rupert 43

The following equation was used to analyze the data that was collected. The t value equals the

mean of the rate of carbon dioxide production in kPa per minute minus the mean rate of carbon

dioxide production in kPa per minute of the second set of data. This is then divided by the

square root of the standard deviation of the first set of data squared divided by the number of

data points in the first set of data plus the standard deviation of the second set of data squared

divided by the number of data points in the second set of data.

x 1 −x 2
t =

(s1 ) 2 (s 2 ) 2
n1 + n2

Figure B2 below shows a sample calculation to determine the p-value.

x 1 −x 2
t =

(s1 ) 2 (s 2 ) 2
n1 + n2

1.648−1.138
= (0.2187) 2 (0.5368) 2
√ 20 + 20

= 3.9348

Figure B2. t-Value of a Two-Sample t-Test

Sample Calculation of a two sample t-test using the first trial, which was comparing Splenda to

Glucose.
Cochill - Quinn - Rupert 44

Figure B3. Calculating Standard Deviation

To calculate the standard deviation, the data set must be first input into column A, which

in Figure B3 above is labeled as dataset. Once all of the data points are entered into column A,

column B needs to be set up so that it subtracts the average value of the data points, by each

individual point, and then squares the difference. For this to have been done correctly, the length

in rows of column A and B should be the same. Column C is then going to be the sum of column

B, which can be found by using the formula sum(heading). However, whatever the column is

called would replace the word heading. Finally, column D will consist of the formula to calculate

the standard deviation of the data set. To do this, take the square root of the sum, or column C,

and divide it by the number of data points in column A. The table on the right side was an

example from the first t-test to calculate the standard deviation of Splenda. This process was

repeated using the Stevia and Glucose data sets.


Cochill - Quinn - Rupert 45

Figure B4. Table C

The figure above shows the table that was used to calculate the p-values after completing

the two-sample t-tests. To acquire the p-value, go to the column labeled “Degrees of Freedom”

and take the number of trials and subtract one. Take the t-value that was calculated using the

equation in Figure B2 and find the closest values within the row of the “Degrees of Freedom”

and go down to the row labeled “One-sided P”. This will give the P value for the t-test.
Cochill - Quinn - Rupert 46

Works Cited

American Diabetes Association. "Statistics About Diabetes." ​American Diabetes

Association​. N.p., n.d. Web. 17 Mar. 2017.

<​http://www.diabetes.org/diabetes-basics/statistics/?referrer=https://www.google.com/​>

Aryal, Sagar. "Glycolysis- 10 Steps Explained Steps by Steps with Diagram." ​Online

Microbiology Notes​. N.p., 08 May 2017. Web. 23 May 2017.

<​http://www.microbiologyinfo.com/glycolysis-10-steps-explained-steps-by-steps-with-di

agram/​>

Buehler, Emily. "Enzymes: The Little Molecules That Bake Bread." ​Scientific American

Blog Network​. N.p., 06 Aug. 2013. Web. 17 Mar. 2017.

<​https://blogs.scientificamerican.com/guest-blog/enzymes-the-little-molecules-that-bake-

bread/​>

Doherty , Jennifer, Dr., and Ingrid Waldron, Dr. "Cellular Respiration In Yeast." ​Serendip

Studio​. University of Pennsylvania Biology Department, 2009. Web. 19 Mar.

2017.

<​http://employee.heartland.edu/hfei/Labs/CellularRespirationProtocol.pdf​>

Editors of Encyclopædia Britannica. "Saccharin." ​Encyclopædia Britannica​.

Encyclopædia Britannica, inc., 20 July 1998. Web. 19 Mar. 2017.

<​https://www.britannica.com/science/saccharin​>

"Fermentation." ​Fermentation | BIOL 011​. The McGraw-Hill Companies, Inc., 2013. Web. 23

May 2017.

<​https://online.science.psu.edu/biol011_sandbox_7239/node/7370​>
Cochill - Quinn - Rupert 47

Jaksich, John A. "Is Commercial Stevia Dangerous? Chemistry, Stevioside and Rebaudioside."

Decoded Science​. N.p., 28 Jan. 2014. Web. 23 May 2017.

<​https://www.decodedscience.org/commercial-stevia-dangerous-chemistry-stevioside-reb

audioside/42111​>

Khan Academy Staff. "Khan Academy." ​Khan Academy​. N.p., n.d. Web. 19 Mar. 2017.

<​https://www.khanacademy.org/science/biology/cellular-respiration-and-fermentation/gl

ycolysis/a/glycolysis​>

<​https://www.khanacademy.org/science/biology/cellular-respiration-and-fermentation/va

riations-on-cellular-respiration/a/fermentation-and-anaerobic-respiration​>

Mayo Foundation for Medical Education and Research. "Video: How diabetes affects

your blood sugar." ​Mayo Clinic​. N.p., n.d. Web. 19 Mar. 2017.

<​http://www.mayoclinic.org/diseases-conditions/diabetes/multimedia/blood-sugar/vid-20

084642​>

Mota, Roxy. "Yeast Fermentation on Natural and Artificial Sugars." ​Prezi.com​. N.p., 03

June 2014. Web. 19 Mar. 2017.

<​https://prezi.com/dheolryvybt8/yeast-fermentation-on-natural-and-artificial-sugars/​>

Science Buddies Staff. "Yeast Reproduction in Sugar Substitutes." ​Science Buddies​.

N.p., n.d. Web. 17 Mar. 2017.

<​http://www.sciencebuddies.org/science-fair-projects/project_ideas/MicroBio_p005.shtm

l#background​>
Cochill - Quinn - Rupert 48

Vernier Software and Technology Staff. "Effect of Temperature on Fermentation." ​Effect

of Temperature on Fermentation > Experiment #16B from Biology with Vernier >

Vernier Software & Technology​. N.p., n.d. Web. 19 Mar. 2017.

<​http://www.vernier.com/experiments/bwv/16b/effect_of_temperature_on_fermentation/

>