Summary of Methodology:

Creatine monohydrate is a protein for muscle mass as it increases it and slows down
muscle breakdown. In this experiment, 5 rats were fed with regular cereal and 5 rats were fed
with cereal dusted with creatine monohydrate powder. Results showed that the rats fed with
creatine monohydrate did increase in weight, indicating increase in muscle mass.
The Effects of Creatine Monohydrate on White Mice (Mus musculus)
Sean Parsa, Heeva Ghane
Department of Biological Science
Saddleback College
Mission Viejo, CA 92692
Creatine is a protein in the form of glycine and arginine. Glycine promotes muscle
building and strength gain by slowing the process of muscle tissue breakdown. Arginine
increases the bodys ability to produce lean muscle mass. The purpose of this study was to
see what effects creatine monohydrate would have on the mass of Mus musculus.
Researchers hypothesized that creatine monohydrate will increase the mass of Mus
musculus. Ten white mice were bought from Wild Animals Supply and were separated into
two groups of five. In the control group, the mice were fed a regular diet of Kelegos
cereal and the experimental group was fed dusted creatine monohydrate Kelegos cereal.
After two weeks, the control increased in weight to 0.010.01g (SEM, n=5) and the
experimental group increased in weight to 0.210.04g (SEM, n=5). These results indicated
that the data obtained did support the researchers hypothesis. If the research was
performed for another consecutive two weeks, the change in weight for the experimental
group would be much greater.
Introduction
Creatine monohydrate has been thoroughly investigated in mammals and was proved to
be a valid performance, body weight, and water volume enhancer. A research done by Ziegenfuss
(1998), tested the acute fluid volume in ten men during three days of creatine supplementation.
Ziegenfusss (1998) subjects were to ingest 0.07 g kg FFM-1 creatine monohydrate dissolved in
500 mL of grape drink every three hours with breakfast, lunch, dinner, and two snacks. This
amount of creatine is approximately 10-20 times that found in a normal diet. Strict dietary
control was observed because changes in nutrition and hydration status could confound estimated
fluid volumes. Specifically, during sessions one, two, and three, subjects completed detailed
dietary records of all ingested foods. The subjects increased in water volume, but each subject
had a different effect on the creatine based on age, weight, and how often they exercised.
Another research done by Vangenberghe (1997) was testing whether creatine supplementation
may add to the effects of resistance training on muscle strength and on the capacity to perform
high intensity exercise and also to evaluate the effects of long-term creatine supplementation on
body composition. Vangenberge (1997) tested this experiment on nineteen women for ten weeks.
The experimental group was given 5g of creatine (2.5g tablets) four times a day. The control
group received placebo supplements (5g of maltodextrine also in tablet form) four times a day.
During the ten weeks, the subjects were to perform variable resistance training for one hour three

times per week. The training involved seven different exercises, including leg press, bench press,
leg curl, leg extension, squat, shoulder press, and sit-ups. In result, creatine increased maximal
strength, maximal intermittent exercise capacity, and fat free mass by 20% to 25%, 10% to 25%,
and 60%. Since the intake of creatine increases the amount of energy produced, the tolerance for
a longer exercise time will increase. Creatine exerts its effect on metabolism by serving as a
precursor to the formation of ATP (Pearlman and Fielding, 2006). When there is an increase in
the amount of creatine present, more ATP will be produced to perform more work (Brink 2005).
Since creatine restores ATP to a state where it can act as a fuel for the muscle, it will enhance
muscle growth. Based upon studies done on humans, the results may be the same on Mus
musculus, since both species are mammals.
Materials and Methods
Ten Mus musculus were bought on October 23, 2009 at Wild Animals Supply, Laguna
Niguel, California. Each mouse was specifically marked using a Sharpie and placed into a
separate container to indicate the experimental group and the controlled group. For 14 days each
mouse was fed five grams of Kellogs Corn flakes cereal every other day. The experimental group
was fed cereal that had been coated with creatine monohydrate. The cereal was dusted with
creatine by spray misting the cereal with water and dusting creatine monohydrate like powdered
sugar over it. To determine the amount of creatine fed, the cereal was weighed before being
dusted with creatine and then weighed after. On the day the mice were fed, they were place into a
0.1778-meter hamster ball to run for five minutes, enabling the effect of the creatine
monohydrate to be enhanced. For the first three days the mice were individually place into a
plastic shoebox (29.5cm x 18cm x 9.5cm) container and then each group of mice was transferred
into a large aquarium. To keep track of how much creatine monohydrate was consumed, each
mouse was placed into separate plastic shoebox containers for about 12 hours every other day to
eat the cereal. Afterward, each mouse was returned to the correct aquarium. The left over cereal
was weighed to see how much creatine had been consumed.
Results
The average change in weight for the control group was 0.010.01g (SEM, n=5). The
average change in weight for the experimental group in this study was 0.210.04g (SEM, n=5).
A one tailed paired t-test revealed that creatine monohydrate does effect the weight of white mice
(p=2.4 x 10-3). These data are shown in Figure 1.

Figure 1. The average weight increase of the controlled and experimental mice. Average weight
increase of the controlled group 0.010.01g (SEM, n=5), experimental weight increase
0.210.04g (SEM, n=5). The average weight increase of the experimental group was
significantly greater than the controlled (p=2.4 x 10-3, one-tailed paired t-test).

12.6

W eigh t (gram s)

12.4
12.2
Control
Experimental
Linear (Experimental)
Linear (Control)

12
11.8
11.6
11.4
11.2
1

Days

Figure 2. The average weight of mice recorded in each group. The experimental group had a
decrease due to excess amount of creatine.

Amount of Food not eaten (grams)

1
0.9
0.8
0.7
0.6
Contol

0.5

Experimental

0.4
0.3
0.2
0.1
0
1

Day

Figure 3. The amount of food the mice did not eat. The experimental group greater amount of
food not ingested due to excess amount of creatine. When the amount of creatine was decreased
the experimental and controlled group had around the same amount of food not eaten.
Discussion
The initial hypothesis was that creatine monohydrate would increase the total body mass
of Mus musculus. The results showed that the creatine monohydrate did increase the total mass
of the mice. The experimental mice lost weight during the beginning of the research period due
to the excess amount of creatine monohydrate that was given, which caused diarrhea. After this
issue was resolved by giving a smaller dose of creatine monohydrate, the mice began to have
increase in weight.
There are many reasons why creatine monohydrate had a positive effect on mice. Mice
have a fast metabolism rate; which may have caused the creatine monohydrate to have a shorter
period of time to pass through the blood stream and distribute throughout the body. The
substance may have a quicker effect in the short period of time because throughout the
experiment the mice were physically active which causes the creatine monohydrate to work
better and faster. In the study done by Ziegenfuss (1998), the subjects were given a large amount
of liquid (500ml of grape drink), because it is said that creatine has a higher effect when the body
is fully hydrate. Throughout the research the mice were given an adequate amount of water in a
cup; which was refilled everyday.
Tythcott (2000) hypothesized that rapid increase in force production might be putting
unwanted stress upon the joints of the Harlan Sprague- Dawley rats. Fourteen 3-4 weeks old
male rats were randomly split-up into separate groups. The experimental group was given
creatine, which was dissolved into a carbohydrate solution (Hawaiian punch fruit drink). Each rat
was given 0.5 cc of a 7.0x10-2 M creatine solution. Each rat received 0.046 g daily for 14 days.
Additional creatine was added to water bottle to ensure administration of minimum dose. As for
the control group, the same amount of Hawaiian fruit punch was given. As for exercising, rats
swam once a day. The study found that supplementation appeared to have an average weight

change in the experimental group. The experimental group experienced a significant average
increase in weight by almost 33%. Tythcott might have had a greater increase in weight because
the creatine monohydrate was also mixed into the fruit punch. Another study done by
Vangenberge (1997) showed that creatine increased maximal strength, maximal intermittent
exercise capacity, and fat free mass by 20% to 25%, 10% to 25%, and 60%. Both of these studies
done by Tythcott and Vangenberge, have the similar procedures and results as our experiment.
This shows that creatine monohydrate does, in fact, increase the weight of mammals and that our
hypothesis is correct.
Acknowledgments
We will like to thank Amir Zand for helping us with choosing the mice for us. We will
also like to thank Mr. and Mrs. Parsa for letting us store the mice at their house throughout the
research. Last, but not least, we will like to thank Professor Teh for supplying us with the
equipment
needed
and
his
guidance.
References
Brink, W. (2005). Creatine Supplementation: Potential Applications in Medicine Townsend.
Letter For Doctors and Patients, 92-95.
Pearlman P., J and Fielding A., R. (Feb 2006). Creatine Monohydrate as a
Therapeutic Aid in Muscular Dystrophy. Nutrition Reviews. 64(2), 80-88.
Powers E. M., Arnold L. B., Weltman L. A., Perrin H. D. Creatine Supplementation increases
total body water without altering fluid distribution. Journal of Athletic Training.38(1), 44.
Tythcott,B.(2000). Effect of Creatine Monohydrate on Tensile Strength of Tendons in Rodents.
Bios, 71(2),35-41.
Vandenberghe K., Goris M. , Van Hecke P., Van Leemputte M. ,Vangerven L., and Hespel P.
(1997). Long-term creatine intake is beneficial to muscle performance
resistance training. Journal of Applied Physiology. 83(6), 2055-2063.
Ziegenfuss N. T., Lowery M. L., and Lemon W.R. P.(October 1998). Acute fluid volume changes
in men during three days of creatine supplementation. Journal of Exercise Physiology.
1(3).