Journal of Applied Microbiology ISSN 1364-5072

ORIGINAL ARTICLE

Microbiological study of used cosmetic products:

highlighting possible impact on consumer health
A. Bashir and P. Lambert
School of Life and Health Sciences, Aston University, Birmingham, UK

Keywords Abstract
Escherichia coli, fungi, microbial
contamination, staphylococci. Aims: To investigate the nature and extent of microbial contamination in five
categories of used cosmetic products (lipstick, lip gloss, eyeliners, mascaras and
Correspondence beauty blenders) and highlight the potential risk posed to consumers in the
Amreen Bashir, School of Life and Health UK.
Sciences, Aston University, Birmingham, UK.
Methods and Results: Used products were donated and microbial contents
E-mail: bashira6@aston.ac.uk
were determined by microbial culture and identification. About 79–90% of all
2019/1410: received 9 August 2019, revised used products were contaminated with bacteria, with bacterial loads ranging
27 September 2019 and accepted 2 October between 102 and 103 CFU per ml, beauty blenders contained an average load
2019 of >106 CFU per ml. Presence of Staphylococcus aureus, Escherichia coli and
Citrobacter freundii was detected. Enterobacteriaceae and fungi were detected in
doi:10.1111/jam.14479 all product types, and were prevalent in beauty blenders (26
58 and 56
96%
respectively). Ninety-three per cent of beauty blenders had not been cleaned
and 64% had been dropped on the floor and continued to be used.
Conclusions: Significant levels of microbial contamination occur during use of
cosmetic products and presence of pathogenic organisms pose a potential risk
to health.
Significance and Impact of the Study: The nature and high level of
contamination in used cosmetic products indicate that greater user awareness
and education are required. Manufacturers should ensure that product expiry
dates are prominently displayed and consumers can identify the symbols used
on product packaging.

cosmetics (Cosmetics Europe – The Personal Care Associ-

Introduction
ation 2019). The demand for imported products in the
Cosmetics are defined by the Federal Food, Drug, and UK highlights the concern that products imported from
Cosmetic Act as ‘articles intended to be rubbed, poured, non-EU markets only have to comply with the legislation
sprinkled, or sprayed on or introduced into, or otherwise of their own jurisdiction.
applied to the human body or any part thereof for The European Cosmetics Regulation ensures cosmetics
cleansing, beautifying, promoting attractiveness or alter- are made to a high standard to ensure consumer safety.
ing the appearance, and articles intended for use as a Testing must be carried out to assess the physical and
component of any such articles; except that such a term chemical characteristics and stability of the product, as
shall not include soap’ (Butler 2000). well as to ascertain the microbiological and toxicological
A recent report identified the EU as having the largest profile (Ec.europa.eu. 2019). US legislation states that
cosmetic market, worth €78
6 billion in 2018. The largest ‘cosmetics are not expected to be totally free of microor-
national markets for cosmetics and personal care prod- ganisms when first used or to remain free during con-
ucts within Europe are Germany (€13
8 billion), France sumer use’ and that ‘cosmetics are not required to be
(€11
4 billion) and the UK (€10
9 billion) stress the sterile, but microbial contamination can pose a health
importance of consumer safety and awareness regarding hazard’ (Food and Drug Administration 2018). EU

598 Journal of Applied Microbiology 128, 598--605 © 2019 The Society for Applied Microbiology
A. Bashir and P. Lambert Microbial contamination of used cosmetic products

guidance states that cosmetics applied around the eye The aim of this study was to investigate the nature and
area should have a total viable count for aerobic micro- extent of microbial contamination in four in-use cos-
organisms no higher than 102 CFU per ml. All other cos- metic products (lipstick, lip gloss, eyeliners, mascaras)
metics should have a total viable count for aerobic and beauty blenders and highlight the potential risk
micro-organisms no higher than 103 CFU per ml. Poten- posed to consumers in the UK.
tially pathogenic species such as Pseudomonas aeruginosa,
Staphylococcus aureus and Candida albicans must not be
Materials and methods
detectable in 1 ml of a cosmetic product applied around
the eye area or in 0
1 ml of other products. EU guidance A total of 467 products from the five makeup categories
also states that the occurrence of Escherichia coli and were donated by product users from the UK, in response
other Enterobacteriaceae is not acceptable in cosmetic to advertisement on social media. This comprised of lip-
products (Scientific Committee on Consumer Safety stick (96), eyeliner (92), mascara (93), lip-gloss (107) and
(SCCP) 2016). beauty blenders (79). Information regarding duration of
Expiry dates of cosmetic products are dictated by the use of each product was collected where possible from an
length of time the preservatives formulated in the pro- associated questionnaire. No personal identifiable data
duct are capable of controlling contamination. If the were collected on product users.
expiry date is printed on the packaging, it is displayed For lipstick and lip gloss products, 1 g of material was
in the form of a symbol resembling an opened pot placed in a sterile universal with 9 ml of phosphate-buf-
with either 3M, 6M, 12M, 18M, 24M or 36M printed fered saline (PBS) containing 10 2.5–3.5 mm glass beads.
in the middle, corresponding to the number of months The used applicator tips from the beauty blender and
the product can be used. A recent study showed that eyeliner product were cut and placed in 10 ml of PBS.
97
9% of participants reported that they use make-up The mascara wands were removed from the product
after the expiration date, with mascara the most fre- tubes and placed in 10 ml of PBS. Each universal was
quently mentioned product. The most common con- mixed by vortex for 1 min and then left to stand for
taminants were S. aureus, found in 79% of samples, 20 min. A 10-fold serial dilution series was performed to
and P. aeruginosa, found in 13% of products (Giacomel reach a final dilution of 10 4. One hundred microlitres
et al. 2013). of each solution was inoculated onto the surface of a
Cosmetics with high water content are at a risk of sup- nutrient agar (NA), mannitol salt agar (MSA), sorbitol
porting microbial growth following in-use contamination MacConkey agar (SMAC), sabouraud dextrose agar
(Lundov et al. 2009 and Birteks€ oz-Tan, T€uys€ € uk
uz, and Ot€ (SAB) and violet red bile glucose agar (VRBGA). SMAC
2013). Use of makeup exposes the products to the com- and VRBGA were incubated for 24 h at 37°C. MSA,
mensal microflora found on the skin, and it is possible whereas NA and SAB were incubated for 48 h at 37°C.
that a low level of contamination may occur at product The number of colonies was counted to calculate CFU
manufacturing level. per ml and colony morphology was observed and
Studies have isolated Salmonella, S. aureus, Staphylococ- recorded. Colonies were characterized using biochemical
cus epidermis, Escherichia coli and P. aeruginosa from tests including catalase, oxidase and Gram staining. The
mascaras, eyeliner and face powder, whilst also identify- identities of selected colonies representative of the popu-
ing the link between S. aureus and conditions such as lations obtained for each product type were confirmed
conjunctivitis and impetigo (Abdelaziz et al. 1989 and using MALDI-TOF MS analysis.
Eldesoukey et al. 2016). Lip glosses and lipsticks have
been identified as vectors for prohibited and pathogenic
Statistical analysis
species such as Escherichia hermannii, S. aureus, Bacillus
cereus and Enterobacter species (Babalola and Eze 2015). Differences in microbial loads were analysed using a
Beauty blenders are recent additions to the cosmetic Kruskal–Wallis one-way analysis of variance and Dunn’s
market and are applicators rather than a cosmetic prod- multiple comparisons test in GRAPHPAD PRISM ver. 7.0.
uct. They are small, synthetic sponges used to blend liq-
uid products such as foundation and concealer into the
Results
skin, therefore have regular contact with the hands and
face. A recent article revealed that in 2016 more than 6
5 Table 1 shows the in-use contamination rates of the five
million beauty blenders had been sold (Shah 2016). product types based upon the counts determined on NA,
Improper use of cosmetic products leading to their con- VRBGA, MSA and SDA. The NA plates showed that
tamination could allow bacteria and fungi to grow, thus approximately 70–90% of all used products were contam-
posing a risk to the consumer. inated with bacteria. Using selective bacterial agar, the

Journal of Applied Microbiology 128, 598--605 © 2019 The Society for Applied Microbiology 599
Microbial contamination of used cosmetic products A. Bashir and P. Lambert

majority of contaminants was found to be staphylococci/ the product types as determined by nonparametric Spear-
micrococci. Enterobacteriaceae were also detected in all man correlation.
product types, with particularly high prevalence in the Only 6
4% of all collected samples had been cleaned,
beauty blenders (26
58%). Fungal contamination was with beauty blenders being the most commonly cleaned
observed at various rates across the product types with product and none of the mascara products. Results
the highest rate in the beauty blenders (56
96%). revealed that 27
3% of products (largely eyeliner) had
Figure 1 shows the microbial loads within each pro- been applied in a bathroom. Consumers were asked
duct group determined on growth medium. Figure 1a whether the product had been dropped on the floor and
shows total bacterial load determined on NA, most used 28
7% of products had being exposed to the floor sur-
products contained bacterial loads between 102 to face. Both these unsanitary practices were observed largely
103 CFU per ml, whereas beauty blenders contained an in beauty blenders, with 35
6% of beauty blender samples
average load of >106 CFU per ml. A one-way ANOVA con- being used or stored in a bathroom, and 64
4% having
firmed that beauty blenders were significantly more con- been dropped on the floor.
taminated than any of the other products tested
(P ≤ 0
003). Figure 1b shows Enterobacteriaceae load
Discussion
determined on VRBGA; lip gloss displayed the highest
load whereas lipstick revealed the lowest load. A one-way Although the effects of consumer use, consumer beha-
ANOVA confirmed that lip gloss was significantly more viour and ineffective preservation have been identified,
contaminated than any of the other products tested there have been few publications from the UK (Dadashi
(P ≤ 0
025). Lipstick was significantly less contaminated and Dehghanzadeh 2016; Eldesoukey et al. 2016). There-
than beauty blenders (P = 0
0229). Figure 1 demonstrates fore, the aim of this study was to determine microbial
bacterial load determined on MSA, the only significant contamination of used products donated by consumers
differences in rates of contamination were detected in the UK.
between beauty blenders and lipstick (P = 0
0184) and The results revealed varying levels of all prohibited
beauty blenders compared to mascara (0
0375). Figure 1 micro-organisms in the used products indicating contam-
demonstrates that the levels of fungal contamination ination caused by the consumer whilst using the prod-
across all product types were approximately 102 CFU per ucts. In many cases, organisms prohibited according to
ml and no significant differences were observed in levels EU guidance in packaged products were present in used
of fungal contamination across the products tested products (Scientific Committee on Consumer Safety
(P ≥ 0
05). (SCCP) 2016). A particular concern is the presence of
Table 2 shows the identity of selected organisms taken potentially pathogenic organisms including; E.coli, C. fre-
from 25 products and confirmed by MALDI-TOF analy- undii, P. aeruginosa and S. aureus. Introduction of these
sis. A total of 48 organisms were identified. Results micro-organisms during application around the mouth
revealed the presence of expected skin organisms, how- or eyes could pose a significant threat of infection. Other
ever, opportunistic pathogens and Enterobacteriaceae studies have revealed the presence of potentially patho-
were also detected. genic micro-organisms in cosmetic products including
Figure 2 shows microbial load vs duration of product Candida species, S. aureus and E. coli (Pascher 1982;
use (months) for individual products where accurate €
Onurdag, Ozgen and Abbasoglu 2010; Dadashi and
dates were available. Logarithmic scales have been used Dehghanzadeh 2016; Eldesoukey et al. 2016).
for the microbial loads on lip gloss and beauty blenders We found P. aeruginosa, P. fulva, P. monteilii and P.
to accommodate the wide numerical ranges obtained for putida in beauty blenders, lipstick and lip gloss. These are
these products. There was no correlation between dura- opportunistic pathogens capable of causing significant
tion of product use and level of contamination for any of infections particularly in immunocompromised

Table 1 In-use contamination rate of products (% of used products found to be contaminated). Four selective agars were used to determine the
presence of organisms in the cosmetic product categories tested

Eyeliner Lipstick Mascara Beauty blenders Lip gloss

n = 92 n = 96 n = 93 n = 79 n = 107

Nutrient agar 80
43 76
04 79
57 92
41 71
96

Violet red bile glucose agar 9
78 5
21 7
53 26
58 9
35
Mannitol salt agar 77
17 56
25 59
14 72
15 55
14
Sabouraud dextrose agar 28
26 36
46 17
20 56
96 10
28

600 Journal of Applied Microbiology 128, 598--605 © 2019 The Society for Applied Microbiology
A. Bashir and P. Lambert Microbial contamination of used cosmetic products

(a) Bacterial load (b) Enterobacteriaceae load

108 104
107

106 103
105
cfu ml–1

cfu ml–1
104 102
103

102 101
101

100 100
k

k
s

s
ra

ra
s

s
rs

rs
tic

tic
os

os
er

er
ca

ca
de

de
in

in
ps

ps
gl

gl
as

as
en

en
el

el
Li

Li
p

p
Ey

Ey
M

M
bl

bl
Li

Li
ty

ty
au

au
Be

Be
(c) Gram-positive load (d) Fungal load
103 103

102 102
cfu ml–1

cfu ml–1

101 101

100 100
k

k
s

s
ra

ra
s

s
s

s
tic

tic
os

os
er

er
er

er
ca

ca
in

in
nd

nd
ps

ps
gl

gl
as

as
el

el
Li

Li
e

e
p

p
Ey

Ey
M

M
bl

bl
Li

Li
ty

ty
au

au
Be

Be

Figure 1 (a–d) The microbial loads within each product group determined on selective growth medium. Enterobacteriaceae levels were detected
using SMAC and VRBGA. Error bars show standard error.

individuals. Although infection can be prevented, ineffec- 1999 and Cone et al. 2005). Staphylococcus haemolyticus is
tive sanitary practices whilst handling products could lead found in the axillae, perineum and inguinal areas (Fis-
to infection via cuts or abrasions in the skin during chetti et al. 2006). Known as an emerging nosocomial
application. Staphylococcus species are commensal organ- pathogen, it has the ability to cause UTIs but is not
isms found on the skin, including S. aureus and Staphylo- known to cause skin infections (Gunn and Davis 1989
coccus epidermidis. In the current study, S. saprophyticus, and Fischetti et al. 2006). Staphylococcus cohnii is a noso-
S. haemolyticus, S. cohnii, and Staphylococcus capitis were comial pathogen, presenting high levels of methicillin
identified through MALDI-TOF analysis from mascara, resistance. It has been known to cause bacteraemia origi-
eyeliner and lip gloss. nating from multiple sources, including pressure ulcers,
Staphylococcus capitis is commonly found on the face UTIs and pneumonia, and has been unusually identified
and therefore expected as a contaminant of cosmetic as the cause of fatal meningitis (Okudera et al. 1991).
products. Staphylococcus capitis infection has been Staphylococcus saprophyticus is commensal to the geni-
reported in endocarditis following pacemaker implanta- tourinary tract but can cause urinary tract infections and
tion and bacteraemia in premature infants (Wang et al. acute cystitis. Although many of these organisms are

Journal of Applied Microbiology 128, 598--605 © 2019 The Society for Applied Microbiology 601
Microbial contamination of used cosmetic products A. Bashir and P. Lambert

Table 2 Identification by MALDI-TOF MS. The identities of 48 isolates were determined. The number of times the isolates were detected in each
product type is listed in parenthesis

Cosmetic product type MALDI-TOF identification of micro-organisms

Beauty blenders Escherichia coli (2) Pseudomonas monteilii (2)

Citrobacter freundii (2) Pseudomonas aeruginosa (2)
Acinetobacter ursingii (2)
Mascara Pluralibacter gergoviae(2)
Staphylococcus saprophyticus (1)
Lipstick Pseudomonas fulva (2)
Pseudomonas monteilii (1)
Citrobacter freundii (1)
Lip gloss Staphylococcus haemolyticus (1) Pseudomonas putida (1)
Staphylococcus cohnii (1) Pseudomonas monteilii (3)
Staphylococcus saprophyticus (2) Pseudomonas fulva (2)
Staphylococcus capitis (2) Lactobacillus (1)
Staphylococcus pasteurii (1) Citrobacter freundii (1)
Micrococcus luteus (1) Candida glabrata (1)
Bacillus litoralis (1)
Eye liner Cryptococcus diffluens (1) Staphylococcus hominis (2)
Micrococcus luteus (1) Staphylococcus haemolyticus (2)
Burkholderia vietnamiensis (1) Escherichia coli (2)
Bacillus muralis (1) Arthrobacter roseus
Staphylococcus saprophyticus (1)
Staphylococcus capitis (2)

commensals to the body, they should not be present in use of symbols to indicate expiry dates (Scientific Com-
products applied to the skin, their presence indicates mittee on Consumer Safety (SCCP) 2016), there are no
poor sanitary practices by the consumer. regulations or requirements under current U.S. laws that
Citrobacter freundii is a Gram-negative species belong- require cosmetic manufacturers to print expiration dates
ing to Enterobacteriaceae family, found in water, soil and on the labels of cosmetic products. However, manufac-
the gastrointestinal tract. This was detected in lipstick turers have the responsibility to determine shelf life for
and lip gloss products. Escherichia coli, a known gut products as part of their responsibility to substantiate
pathogen was also observed across all product types and product safety (Food and Drug Administration 2019).
confirmed in beauty blenders and eyeliner products by Sharing makeup products and makeup testers found
MALDI-TOF. The cross-contamination of cosmetic prod- on beauty counters, may also provide a route for con-
ucts with both of the organisms indicates poor consumer tamination and infection. Commonly testers are not
hygiene whilst handling and applying products. cleaned regularly, and are left exposed to the environ-
Beauty blenders have only been recently introduced as ment and to passing customers who are allowed to touch
an application product and limited information is avail- and try the product. Sharing makeup greatly increases the
able on how best to use or clean them. Our results have level of contamination seen in testers due to the increased
shown that these products carried the highest bacterial use and pressure on cosmetic preservatives, and this can
load during use and more than a quarter were contami- include contamination by bacteria, fungi and yeast
nated with Enterobacteriaceae. Acinetobacter ursingii was (Dadashi and Dehghanzadeh 2016).
also detected in this product type, a Gram-negative coc- All cosmetic products are manufactured under strictly
cobacillus known as an important opportunistic pathogen controlled conditions to control microbial content and
known for causing nosocomial infections. Beauty blenders proliferation during use. The lifetime of the product is
could pose a significant risk to consumers, their product limited by the protection afforded by the preservation
design allows micro-organisms to accumulate. Beauty system and this information is provided on the label of
blenders can be cleaned with warm, soapy water therefore the product. Most products have an expiry date of 3–
encouraging microbial proliferation if not dried. 12 months, providing the user has not had an infection
Many products are imported from other countries and such as conjunctivitis. However, our study shows in prac-
are not obliged to provide expiry information on product tical terms products are used beyond the expiration date
packaging. Although EU guidance has incorporated the of the products and considerable levels of microbial

602 Journal of Applied Microbiology 128, 598--605 © 2019 The Society for Applied Microbiology
A. Bashir and P. Lambert Microbial contamination of used cosmetic products

(a) Eyeliner (b) Mascara

600 4000

3000
on NA (cfu ml–1)

on NA (cfu ml–1)
Microbial load

Microbial load
400

2000

200
1000

0 0
0 50 100 150 200 0 20 40 60 80
Age of product (months) Age of product (months)

(c) Lipstick (d) Lip gloss

2500 105

2000 104
on NA (cfu ml–1)

on NA (cfu ml–1)
Microbial load

1500 Microbial load 103

1000 102

500 101

0 100
0 50 100 150 0 50 100 150
Age of product (months) Age of product (months)

(e) Beauty blenders

109
108
107
on NA (cfu ml–1)
Microbial load

106
105
104
103
102
101
100
0 20 40 60 80
Age of product (months)

Figure 2 (a–e) Microbial load plotted against the age of products.

contamination have been identified across a range of cos- sharing of cosmetic products or factors such as proce-
metic products. dures used to clean beauty blenders between use.
The microbial levels and frequency of contamination Cosmetic regulations clearly state that products should
in used products revealed in this study suggest that fur- not contain pathogenic organisms. About 70–90% of all
ther detailed investigations should be carried out to used products were contaminated with bacteria. The
ensure better user compliance. Future studies should presence of organisms including S. aureus, E. coli and C.
incorporate effective preservative utilization steps and freundii in used products may be introduced through
anaerobic culture should be included. This study has not ineffective hand hygiene. Information derived from our
considered the effect on microbial contamination of questionnaires showed that 93% of beauty blenders had

Journal of Applied Microbiology 128, 598--605 © 2019 The Society for Applied Microbiology 603
Microbial contamination of used cosmetic products A. Bashir and P. Lambert

not been cleaned and 64% had been dropped on the endocardial pacemaker infection: case report and review of
floor and continued to be used. Enterobacteriaceae were Staphylococcus capitis endocarditis. Int J Infect Dis 9,
also detected in all product types, with particularly high 335–339.
numbers observed in the beauty blenders (26
58%). Fun- Cosmetics Europe – The Personal Care Association. (2019).
gal contamination was observed across the product types Cosmetics Industry. [online] Available at: https://cosme
with the highest rate detected in the beauty blenders ticseurope.eu/cosmetics-industry (accessed 8 Aug 2019).
(56
96%). Clearly further advice and education are Dadashi, L. and Dehghanzadeh, R. (2016) Investigating
needed on the use and maintenance of these products to incidence of bacterial and fungal contamination in shared
cosmetic kits available in the women beauty salons. Health
avoid self-contamination with potentially harmful micro-
Promotion Perspect 6, 159–163.
organisms. Such potential hazards would be increased
Ec.europa.eu. (2019). [online] Available at: https://ec.europa.e
where products are used by multiple individuals such as
u/health/sites/health/files/endocrine_disruptors/docs/cosme
in beauty salons. We are not aware of any regulations or
tic_1223_2009_regulation_en.pdf (accessed 8 August
discussion groups addressing contamination issues of
2019).
beauty blenders, which are applicators used to apply cos- Eldesoukey, R.M.M., Alqhtani, B.S., Alqhtani, A.S., Alqhtani,
metics rather than a cosmetic product incorporating a A.H. and Alqhtani, A.M. (2016) Comparative
preservation system. Manufacturers also need to ensure microbiological study between traditional and modern
that product expiry dates are prominently displayed and cosmetics in Saudi Arabia. Enzyme Eng 5 https://doi.org/
consumers can identify the symbols used on product 10.4172/2329-6674.1000146.
packaging. Fischetti, V., Novick, R., Ferretti, J., Portnoy, D. and Rood, J.
(2006) Gram-Positive Pathogens (2nd edn) pp. 572–586.
Washington D.C: ASM Press.
Acknowledgements
Food and Drug Administration. (2018). Small Businesses &
We would like to thank the following individuals who Homemade Cosmetics: Fact Sheet. [online] Available at:
contributed to the acquisition of the data; Isabelle https://www.fda.gov/cosmetics/resources-industry-cosmetic
Adams, Sarah Adesina, Selina Ashfaq, Rebecca Ellender, s/small-businesses-homemade-cosmetics-fact-sheet
Rachel Hepton, Nathalie Lenders, Ethel Ojo, Ugne Pau- (accessed 8 Aug. 2019).
lauskaite, Eleanor Sadler and Jessica Willett. We would Food and Drug Administration (2019). Do I need to label my
products with expiration dates? [online] U.S. Food and
also like to thank the microbiology department at Birm-
Drug Administration. Available at: https://www.fda.gov/
ingham children hospital for identification of isolates on
industry/fda-basics-industry/do-i-need-label-my-products-
the MALDI-TOF MS.
expiration-dates (accessed 8 Aug. 2019).
Giacomel, C., Dartora, G., Dienfethaeler, H. and Haas, S.
Conflict of Interest (2013) Investigation on the use of expired make-up and
microbiological contamination of mascaras. Int J Cosmet
The authors have no conflict of interest to declare. Sci 35, 375–380.
Gunn, B. and Davis, C. (1989) Staphylococcus haemolyticus
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Journal of Applied Microbiology 128, 598--605 © 2019 The Society for Applied Microbiology 605