Running head: THE EFFECTS OF BACTERIA AND OIL 1

THE EFFECTS OF BACTERIA AND OIL’S RELATIONSHIP IN COSMETIC

FOUNDATIONS ON THE PRODUCTION OF FACIAL ACNE AS SEEN IN CONSUMER

REVIEWS

By

Natalie Pachucki

A Senior Thesis Submitted in Partial Fulfillment of Graduation Requirements

Central Magnet School

May 2018

Thesis Committee:

Dr. Jeff Leblond

Adam Brooks

Eve Harrison

Laura Lynn Roland

THE EFFECTS OF BACTERIA AND OIL 2

Acknowledgements

First, I would like to thank Dr. Jeff Leblond for serving as my mentor for my senior

thesis project. Without his flexibility and extensive scientific knowledge, my thesis would not

have been possible. I would also like to thank Mr. Adam Brooks, who has served as my advisor

for the past four years. Without his knowledge and willingness to help, I wouldn’t have been able

to make it this far.

Next, I would like to thank my Biomedical Sciences teacher, Mrs. Eve Harrison. Without

her guidance and help, my senior thesis would not have come together as smoothly as it did.

Finally, I would like to thank Mrs. Laura Lynn Roland. Her enthusiasm and excitement everyday

gave me the motivation to push through all of my assignments. Her knowledge greatly helped to

improve my thesis, and nothing this year would have been possible without her help.
THE EFFECTS OF BACTERIA AND OIL 3

Table of Contents

Title Page 1

Acknowledgements 2

Table of Contents 3

Abstract 5

Introduction 6

Research Question 6

Research Purpose 6

Background Information 6

Bacteria 6

Staphylococcus Aureus 7

Staphylococcus Epidermidis 7

E.Coli 8

Oil 8

Bacteria and Oil’s Relationship 9

Acne and Human Skin 9

Hypothesis 10

Methodology 11

Materials 11

Procedure 11

Measures 13

Images 1-3 14
THE EFFECTS OF BACTERIA AND OIL 4

Results 15

Limitations 21

Conclusion 21

References 23
THE EFFECTS OF BACTERIA AND OIL 5

Abstract

The purpose of this experiment is to determine if the presence of oil and bacteria in

specific cosmetic foundations are dependent on one another, and if these levels are related to the

production of facial acne as seen in consumer reviews. Based on prior research, it was

hypothesized that oil and bacteria would be dependent on one another, oil levels rising in the

absence of bacteria. It was also hypothesized that acne levels would be reported more frequently

in the foundations containing higher oil contents. Upon results, it was clear to see that no

bacterial colonies were formed during the experimental phase. While analyzing bad reviews and

determining the percentage of oil levels in each foundation, a correlation was found. Therefore, it

was uncertain whether or not bacteria and oil levels were dependent on one another. However, it

was suggested that oil levels being higher in cosmetics leads to more negative reviews. Further,

more accurate, research may need to be done in the future to get a better conclusion.
THE EFFECTS OF BACTERIA AND OIL 6

Introduction

Research Question

Is the amount of bacteria present in cosmetic foundations dependent on the oil levels

measured in the same cosmetics? How does this relate to the production of facial acne as

observed in consumer reviews?

Research Purpose

The purpose of my experiment is to determine the levels of oils and bacteria present in

specific used cosmetic foundations in an attempt to see if they are dependent on one another. The

intent of this study is to find a correlation between oil and bacteria in two specific cosmetic

foundations in order to determine whether or not these levels contribute to skin acne. Naturally,

there are other variables that contribute to facial acne, but the main study focus is centered on

bacterial populations. I will be testing Maybelline FIT Me Stay Matte Foundation and

Maybelline FIT Me Dewy Foundation.

Background Information

Bacteria

Bacteria is defined as a disease carrying microorganism that can be found virtually

everywhere. A common place in which many forget bacteria readily grows is on or in cosmetic

makeup products. Cosmetics such as lipstick, foundation, applicator sponges, and eyeshadow

palettes all harbor multiple types of bacteria. The most common growths on cosmetics, as

observed by Aakriti Guleria, a researcher from Himachal Pradesh, India, include ​Staphylococcus

and ​E.coli.
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Staphylococcus Aureus

Staphylococcus Aureus ​is a Gram-positive microorganism. It​ ​is said to be found in the

noses of about 30% of the population. While this type of bacteria does not cause harm, it can

sometimes lead to infections. Especially in healthcare settings, such as hospitals, these infections

can prove fatal. In a study done in Valencia, a team of microbiologists ran a test to determine the

presence or absence of ​Staphylococcus aureus ​in 1 gram of a cosmetic sample. The cosmetic

sample was diluted in a 1:10 ratio and incubated for 24-72 hours at 32.5°C. After this process

was completed, the samples were observed and identified as ​Staph. aureus.

In another study done by Edlira Neza and Marisanna Centini at the University of Siena in

Siena, Italy, the Rapid Alert System was used to find information about dangerous cosmetic

products sold in EU markets. They defined cosmetics as products that are recognized to be

beneficial for the survival and development of many microorganisms. Studies have found that ​S.

aureus ​is one of the most common microorganisms found in cosmetics. In order for ​S. aureus ​to

be found unharmful, it cannot be detected in 0.5g or 0.1 mL of the product. In Neza’s and

Centinis study, mostly face products such as creams and moisturizers were found to be

contaminated with ​S. aureus. ​It was found most commonly in shea butter and massage cream as

well as other cosmetics.

Staphylococcus Epidermidis

Just like ​S. aureus, S. Epidermidis ​is a gram-positive bacteria that is commonly found in

many cosmetics. ​ ​However, unlike ​S. aureus, ​this bacteria is proven to be beneficial to the skin

due to its apparent moisturizing properties (AMI, 2015). In this study, ​S. Epidermidis ​was

removed from samples of patients’ forehead skin. The bacteria was cultured and then freeze
THE EFFECTS OF BACTERIA AND OIL 8

dried in order to preserve it. The cultured bacteria was then added to face gel and applied to the

patients’ faces. Patients who received the gel were shown to have 1.4 times the amount of water

in their skin after the trial ended.

E.coli

E.coli ​is defined as a type of bacteria that is commonly found in the intestines of humans

and other animals. In the cosmetic world, ​E.coli ​normally gets brought up in relation to makeup

testers (Brooks, 2010). Dr. Elizabeth Brooks at Jefferson Medical College in Pennsylvania

describes the word ​E.coli ​as being synonymous to faeces, meaning someone used the restroom

and didn’t wash their hands, transferring those germs to whatever they next touched. In her

study, she tested different makeup testers from various department stores and found various

types of bacteria harboring in them, mainly ​E.coli.

Oil

In the majority of cosmetic products on the market, different types of oils can be seen as a

major ingredient. The most common types of oil found in these products are mineral oil and

vegetable oil. Mineral oil is widely used based off of its clear and odorless consistency. It has

been proven to rarely cause allergic reactions and cannot become solid and clog pores. Vegetable

oil is used due to its ability to create a barrier on the skin’s surface, slowing down the loss of

water. This allows the skin to stay moist and “plump.”

The problem with using mineral oils stems from the fact that it is a nonrenewable

resource, making the process of extraction difficult. The other issue comes from the impurities

that can be found in mineral oil upon extraction. In an effort to fight hazardous levels of bacteria
THE EFFECTS OF BACTERIA AND OIL 9

from growing, mineral oil and vegetable oil have both been refined to remove mold, fungus, and

bacteria.

Bacteria and Oil’s Relationship

Mineral oil is said to be derived from petroleum oil. Mineral oil can be identified in many

facial products, including foundations, moisturizers, and other makeup products. While some

mineral oils have been heavily refined in order to remove any types of bacteria, as stated above,

there is a great chance that some have not. In a 2011 study described by Dr. Frank Lipman,

“There is strong evidence that mineral oil hydrocarbons are the greatest contaminant of the

human body, amounting to approximately 1 gram per person.” These hydrocarbons come about

from the impurities not being taken out of the sample oils. When not refined, these mineral oils

can grow bacteria, causing clogged pores and acne breakouts among cosmetic users (Lipman,

2014).

Acne and Human Skin

Acne is defined as the occurrence of inflamed or infected sebaceous glands in the skin.

Studies from around the world have all concluded various reasons for the occurrence of facial

acne. Among these include diet, hygiene, genetics and environmental factors. However, it can

take anything as small as an imbalance in simple microbiota to cause acne-causing bacteria to

flourish. Cosmetics have been proven to stimulate the growth of acne due to their pore-clogging

properties.
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Hypothesis

Based on the research above, it is believed that oil and bacteria levels found in cosmetics

will be dependent on one another. I hypothesize that the higher the oil levels are in each product,

the lower the bacteria levels will be. It is also believed that these factors will have a negative

effect on the development of facial acne. According to previous research, cosmetics contain

naturally occurring oils and bacterial colonies, supporting this hypothesis.

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Methodology

Materials

Before beginning the experiment, four cosmetic foundations were purchased from the

store. The foundations purchased included Maybelline FIT Me Stay Matte Foundation, L’Oreal

Paris Infallible Pro-Glow Foundation, Revlon ColorStay Makeup Foundation, and Neutrogena

Healthy Skin Liquid Makeup Foundation. Before being bought, each foundation was checked to

insure the seal had not been broken.

These foundations were chosen based on the number of their sales to ensure that the

variety of consumer reviews would be plentiful. Each sample was diluted by 1% peptone water.

The selected dilutions were spread-plated on Plate Count Agar (PCA). Selected dilutions will

also be spread-plated on Baird-Parker Agar, Mannitol Salt Agar, Eosin Methylene Blue Agar,

and Pseudomonas Isolation Agar. The media's will then be incubated for 1-2 days.

Procedure

After each foundation obtained, Maybelline FIT Me Stay Matte Foundation, L’Oreal

Paris Infallible Pro-Glow Foundation, Revlon ColorStay Makeup Foundation, and Neutrogena

Healthy Skin Liquid Makeup Foundation samples were decimally diluted in 1% peptone water.

This was chosen because peptone water is an all-purpose diluent in bacterial plate counting

methods. After being diluted, selected samples were spread-plated on Plate Count Agar, known

as PCA. The PCA was chosen because it is a medium typically used in total plate count

procedures in order to estimate heterotrophic bacteria in terms of colony forming units per unit

of substance (g or ml).
THE EFFECTS OF BACTERIA AND OIL 12

Additionally, selected dilutions were also spread-plated on four selective media in order

to determine targeted bacterial populations. The first media used was Baird-Parker Agar, which

is a media designed for the detection of ​Staphylococcus ​spp. S. ​aureus ​forms black and shiny

convex colonies surrounded by clear zones. Other species produce gray or slightly less shiny

black colonies. Mannitol Salt Agar (MSA), a selective-differential medium used to isolate

Staphylococcus ​spp., was also used. MSA contains 7.5% NaCl, which chooses against other

bacteria, specifically Gram-negative bacteria. It also contains a pH-sensitive dye, phenol red, and

the carbohydrate, mannitol. When mannitol is fermented to lower the pH of the medium, it turns

from red to yellow. Next, Eosin Methylene Blue Agar, a solid selective and differential medium

used at completed level, was used. EMB, when at acidic pH levels, reacts to form a dark purple

precipitate. Some strains of ​Escherichia coli ​may even ferment so much that the dyes turn a

metallic green. Non-lactose colonies would appear colorless and pale colored. Sucrose

fermenters would appear pink. The last medium used was ​Pseudomonas ​Isolation Agar, which is

a selective medium for the isolation of pseudomonads. It uses glycerol as a source of carbon and

energy. It contains Irgasan, a broad-spectrum antimicrobial agent; magnesium chloride and

potassium sulfate to enhance the production of pyocyanin, a fluorescent pigment produced by

Pseudomonas aeruginosa. ​The media was then incubated for 1-2 days at 37°C. After incubation,

each sample was evaluated for the levels of bacteria produced. After the levels of bacteria

produced were evaluated, an analysis of key ingredients was done. Each list was then compared

to determine any common active ingredients. Oil levels were then determined and analyzed

through calling the company who produced the cosmetic foundation. These levels were recorded.

Consumer reviews were then read and analyzed. The reviews were separated based on reports of
THE EFFECTS OF BACTERIA AND OIL 13

acne breakouts. A correlation was then determined between bacteria growth and oil contents in

the foundations. Reports of acne breakouts were then reexamined and correlated to the

experimental results.

Measures

In order to measure the levels of bacteria produced by each foundation, bacterial colonies

were counted and recorded from each agar plate. Oil levels were measured using the ingredients

list located on the back of each foundation package. The amounts of facial acne related to each

foundation were measured and determined by online consumer reviews. Like the oil levels, the

ingredients were determined by the ingredients list on the back of each passage.
THE EFFECTS OF BACTERIA AND OIL 14

Images 1-3

(image 1) (image 2) (image 3)

Image 1: ​Labeled agar plates that were placed in the incubator after samples were spread on
them
Image 2: ​Close-up of labeled agar plates
Image 3: ​Diluted foundation test tubes
THE EFFECTS OF BACTERIA AND OIL 15

Results

Revlon Colorstay Foundation (unopened)

Results for bacterial colony growth in each dilution:

(Same results for all Agar plates)

Dilution 1 2 3 4 5 6

10​-1 0 0 0 0 0 0

10​-2 0 0 0 0 0 0

10​-3 0 0 0 0 0 0

10​-4 0 0 0 0 0 0

10​-5 0 0 0 0 0 0

10​-6 0 0 0 0 0 0
(Figure 1)

L’Oreal Paris Infallible Pro-Glow Foundation (unopened)

Results for bacterial colony growth in each dilution:

(Same results for all Agar plates)

Dilution 1 2 3 4 5 6

10​-1 0 0 0 0 0 0

10​-2 0 0 0 0 0 0

10​-3 0 0 0 0 0 0

10​-4 0 0 0 0 0 0

10​-5 0 0 0 0 0 0

10​-6 0 0 0 0 0 0
(Figure 2)
THE EFFECTS OF BACTERIA AND OIL 16

Covergirl Clean Makeup (unopened)

Results for bacterial colony growth in each dilution:

(Same results for all Agar plates)

Dilution 1 2 3 4 5 6

10​-1 0 0 0 0 0 0

10​-2 0 0 0 0 0 0

10​-3 0 0 0 0 0 0

10​-4 0 0 0 0 0 0

10​-5 0 0 0 0 0 0

10​-6 0 0 0 0 0 0
(Figure 3)

Maybelline FIT Me Matte Foundation (used)

Results for bacterial colony growth in each dilution:

(Same results for all Agar plates)

Dilution 1 2 3 4 5 6

10​-1 0 0 0 0 0 0

10​-2 0 0 0 0 0 0

10​-3 0 0 0 0 0 0

10​-4 0 0 0 0 0 0

10​-5 0 0 0 0 0 0

10​-6 0 0 0 0 0 0
(Figure 4)
THE EFFECTS OF BACTERIA AND OIL 17

Maybelline FIT Me Dewy Foundation (used)

Results for bacterial colony growth in each dilution:

(Same results for all Agar plates)

Dilution 1 2 3 4 5 6

10​-1 0 0 0 0 0 0

10​-2 0 0 0 0 0 0

10​-3 0 0 0 0 0 0

10​-4 0 0 0 0 0 0

10​-5 0 0 0 0 0 0

10​-6 0 0 0 0 0 0
(Figure 5)

After diluting each unopened foundation sample and vortexing the contents inside to

ensure equal distribution, the samples were each spread across various types of plate agar. They

were then incubated for a full 24 hours. The next day, upon examination, there were no bacterial

colonies detected. The first experimental process was done using four unopened cosmetic

foundations. Upon the results, I decided to redo my experiment, changing the foundation types

and making sure they were used. I started with an opened bottle of Maybelline FIT Me Matte

Foundation and an open bottle of Maybelline FIT Me Dewy Foundation. I repeated the above

process: diluting the samples, vortexing them, and then spreading them on various agar plates.

After 24 hours of incubation, it was then determined that no bacterial colonies had grown.
THE EFFECTS OF BACTERIA AND OIL 18

After measuring the amounts of bacterial colony growth in each foundation, the oil levels

of each opened cosmetic were determined. By calling the Maybelline company, I was able to

determine what percentage of oil was present in each foundation. I found that the Dewt

foundation contained approximately 3% of mixed oils and the Matte foundation contained

approximately 1% of mixed oils. I was unable to determine the kinds of oils in each product,

despite my research.

After measuring the oil levels, I went to Maybelline.com and Ulta.com to analyze the

reviews on Maybelline FIT Me Matte Foundation and Maybelline FIT Me Dewy Foundation. I

recorded any reports of breakouts for each foundation, as well as any reports of large amounts of
THE EFFECTS OF BACTERIA AND OIL 19

oil being present after application. After analyzing the results, I copied my information into a

chart, as seen below.

THE EFFECTS OF BACTERIA AND OIL 20
THE EFFECTS OF BACTERIA AND OIL 21

Limitations

In my experiment, there proved to be an abundance of limitations that arise throughout

the experimental process. To begin with, the samples of the foundations being diluted and spread

across the agar plates that I tested were not equal. Each pipette was filled with 1mL of the

cosmetic, but on occasion certain agar plates got more product than others. Another limitation

may have come from the fact that the foundations I used were previously owned and used by me

and I had no recollection on the age of the products. Therefore, my negative results may have

been directly caused by the age of the products. Lastly, the agar that was used to grow the

bacterial colonies may not have been suitable for the growth of the foundation’s properties. This

may have also caused my results to be negative.

As for the oil measurements, I was unable to complete the results by my own experiment.

Therefore, the results may have been slightly off or invalid due to different websites and

information sources.

Conclusion

The results of this study have shown that the levels of bacteria and oil in these two

specific foundations are not directly related. The presence of oil in the Dewy foundation was 3%,

while the presence of oil in the Matte foundation was 2%. In correlation to the consumer reviews

I analyzed, it was easy to see that the Dewy foundation contained more reviews that expressed

high oil levels and varying breakouts. The Matte foundation experienced reviews that expressed

the same results, but to a slightly smaller degree.

Based on this, it can be concluded that higher oil levels may cause higher bacterial

growth due to the comments analyzed in the consumer reviews. Although no bacterial colonies
THE EFFECTS OF BACTERIA AND OIL 22

were found on the agar plates after incubation, the oil levels examined correspond with the

consumer reviews, supporting the hypothesis that more oil leads to more bacteria, creating a

dependent relationship among the two.

THE EFFECTS OF BACTERIA AND OIL 23

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El Romeral, M. (2016). Cosmetics testing - Microbiology of cosmetics. Detection of

Staphylococcus aureus (UNE-EN ISO 22718: 2016) (Accredited by ENAC). Retrieved

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http://www.ivami.com/en/microbiology-of-cosmetics-laboratory-of-control-authorized-b

y-aemps-8-accredited-tests/3357-cosmetics-testing-microbiology-cosmetics-detection-i-st

aphylococcus-aureus-i-une-en-iso-22718-2016-b-enac-test

Fit Me Dewy & Smooth Foundation - Foundation for Dry Skin. (2017, April 23). Retrieved

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sku=2225225&cmpid=PS_Non%21google%21Product_Listing_Ads&cagpspn=pla&CA

TCI=aud-334903466095%3Apla-195006210030&CAAGID=38553845491&CAWELAI

D=772017114&catargetid=330000200000805942&cadevice=c&gclid=CjwKCAiA8bnU

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oCsL4QAvD_BwE

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ndation/natural-tan
THE EFFECTS OF BACTERIA AND OIL 24

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u1v7EbvUiRYhoCvxIQAvD_BwE

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https://www.cdc.gov/hai/organisms/staph.html

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Oakes, K. (2017, October 15). The microbiota matters: In acne, it's not us versus them. ​Family

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d20709b

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THE EFFECTS OF BACTERIA AND OIL 25

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