BACKGROUND:

Many women tend to keep makeup past the recommended time they are supposed to definitely
when the makeup costs over $50. Old makeup is the prime breeding ground for bacteria. It is rare
for makeup to cause infection and irritation, but it is still very possible. Eye makeup is the most
known to do this. It can harbor bacteria that can cause conjunctivitis, or more commonly known
as pink eye. The FDA doesnt require companies to put expiration dates on beauty products;
although, many dermatologists suggest times when to throw away certain makeup products.
Products that will most likely cause infection are products that are dark and damp. For example,
makeup that is cream or liquid such as mascara. Makeup contains preservatives that should kill
common-use bacteria. But, as studies have shown, a little bacteria is usually present in makeup
before it is bought. Then, by opening and touching the product with unclean hands or an unclean
brush, it exposes the product to bacteria.The method of application is a major factor of how
much bacteria will end up on a product. The applicator, be it your hands or a brush, if unclean
can bring pathogens onto your face.
Problem Statement:
Will makeup grow bacteria so that you have to put a time restriction on how long it should be
used, or is the the method of the application what causes bacteria growth?
Hypothesis:
The oldest and most moist product, such as the mascara, will have the most bacterial growth on
it.
Materials:

New, unused makeup

dry foundation
mascara
lipstick
Makeup samples from my house
dry foundation
mascara
lipstick
Inoculating loops
Agar solution
Petri dishes
Microfuge tubes
Nutrient broth
Pipets
Micropipet

 Box

Procedure:
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.

Gather makeup samples from the house

Gather makeup samples from control (unused, new makeup)
Prepare the agar solution. In a heatproof dish, stir in agar powder into 60 mL of water
Place the dish into the microwave for a minute, make sure it doesnt boil over.
Pour the agar solution into the sterile petri dishes and let it cool.
Using a sterile scoop, scoop up a small bit of the makeup sample and place it into a microfuge
tube and make sure to label the tube with the makeup thats in it.
Repeat step six with the different types of makeup.
Get a box and poke 30 holes in it for all the microfuge tubes. There should be 6 rows and 5
columns. The makeup samples should be in the first row.
Use a pipet to measure 0.1 ml of nutrient broth into all the microfuge tubes. Leave the tubes
slightly pre-opened so itll be easier to fill.
To mix the sample and broth together, tap the end of each microfuge tube to combine the makeup
and the broth. DONT SHAKE THE TUBE.
Now, take the pipet that measure 100 microliters. Attach a pipet tip. Pipet microliters of liquid
from the first microfuge tube to the second.
Using the same pipet tip, move the liquid from the second tube to the third and continue until the
last one in the column is reached.
Repeat steps 10-12 for the other 5 samples.
Now obtain the petri dishes and inoculating loops.
Using the 100 microliters pipet, and a sterile pipet tip, measure 200 microliters of liquid from the
second microfuge tube from the first row onto a petri plate.
Use an inoculating loop to spread it.
Close the lid and label it.
Discard the cotton swab and pipet tip
Using a fresh petri dish, inoculating loop, pipet tip, repeat steps 14-18 with the other microfuge
tubes with the samples.
Tape all the lids shut
Label the dishes with the appropriate number and stack them.
Place them in a safe area at room temperature and colonies will form in 24-48 hours.
After 24-48 hours check on the petri dishes and look if there is any growth visible.
Place over a light and count the colonies. Use a sharpie to keep track of the colonies that have
been counted.
If there is growth visible, then count the number of bacterial colonies on each agar plate and
record your data.
Data:
Day 1:

Name of Sample

Number of Bacterial Colonies

Dry foundation

none

Lipstick

none

Eyeshadow

none

Mascara

none

Control

none

.Day 2:

Name of Sample

Number of Colonies

Dry foundation

none

Lipstick

none

Eyeshadow

none

Mascara

none

Control

none

Day 3:
Name of Sample

Number of Colonies

Dry foundation

none

Lipstick

none

Eyeshadow

Mascara

none

Control

none

Day 4:

Name of Sample

Number of Colonies

Dry foundation

none

Lipstick

none

Eyeshadow

Mascara

none

Control

none

Day 5:

Name of Sample

Number of Colonies

Dry foundation

none

Lipstick

none

Eyeshadow

Mascara

none

Control

none

During day 1 there was no bacterial growth in any of the agar plates. The same occurred during
day 2. Then, finally on day 3-5 there was bacterial growth on the agar plate that contained the
eye shadow. Although, there was no growth in the others.
Analysis:
The only agar plate that contained any formation of bacterial growth was the sample that
contained the eyeshadow. That means that there was a bacteria present in that sample. The
control in all five days had no bacterial growth which was expected since that there was no
sample placed on that plate. The others also did not exhibit any growth. This must mean that
there was no bacteria present in the samples.
Conclusion:
The only sample that contained bacteria was the eyeshadow. This was surprising because a dry
surface is not prime living conditions for bacterial growth. I was quite surprised that the mascara

also did not exhibit any growth in the sample. I expected the mascara to have the most. This is
because the mascara tube is damp and dark, both conditions are perfect for bacteria to grow in.
The samples I gathered may not have been used as much and if I were to run this experiment
again, I would gather samples from products that look like they have been used the most.