Journal Pre-proof

Controversy around parabens: alternative strategies for preservative use in cosmetics

and personal care products

Karolina Nowak, Ewa Jabłońska, Wioletta Ratajczak-Wrona

PII: S0013-9351(20)31385-2
DOI: https://doi.org/10.1016/j.envres.2020.110488
Reference: YENRS 110488

To appear in: Environmental Research

Received Date: 26 May 2020

Revised Date: 9 November 2020
Accepted Date: 12 November 2020

Please cite this article as: Nowak, K., Jabłońska, E., Ratajczak-Wrona, W., Controversy around
parabens: alternative strategies for preservative use in cosmetics and personal care products,
Environmental Research, https://doi.org/10.1016/j.envres.2020.110488.

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 Controversy around parabens: alternative strategies for preservative use in cosmetics
and personal care products
Karolina Nowak1*, Ewa Jabłońska1, Wioletta Ratajczak-Wrona1

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1 – Department of Immunology, Medical University of Bialystok, Poland.

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* Corresponding author: Karolina Nowak
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Medical University of Bialystok

Department of Immunology
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Waszyngtona 15A,
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15-269 Bialystok, Poland

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Phone: +48 85 745 05 47

E-mail: karolina.nowak@umb.edu.pl

Karolina Nowak ORCID: 0000-0002-9971-0023

Ewa Jabłońska ORCID: 0000-0001-7899-7830

Wioletta Ratajczak-Wrona ORCID: 0000-0002-7634-0875

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Abstract

Parabens usage as preservatives in cosmetics and personal care products have been debated

among scientists and consumers. Parabens are easy to production, effective and cheap, but its

safety status remains controversial. Other popular cosmetics preservatives are formaldehyde,

triclosan, methylisothiazolinone, methylchloroisothiazolinone, phenoxyethanol, benzyl

alcohol and sodium benzoate. Although their high antimicrobial effectiveness, they also

exhibit some adverse health effects. Lately, scientists have shown that natural substances such

as essential oils and plant extracts present antimicrobial potential. However, their use in

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cosmetic is a challenge. The present review article is a comprehensive summary of the

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available methods to prevent microbial contamination of cosmetics and personal care
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products, which can allow reducing the use of parabens in these products.
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Keywords
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parabens; cosmetics; preservatives; self-preserving cosmetics; personal care products

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This research did not receive any specific grant from funding agencies in the public,

commercial, or not-for-profit sectors.

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Highlights

• Regarded to adverse health effects parabens are gradually replaced by other compounds

• Natural compounds have a potential for being used as preservatives

• Good Manufacturing Practices in cosmetics ensure the quality of product

• Novel strategies of preservation enable safe and efficient prevention against

microbiological contamination

Abbreviations

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ADHD – attention deficit hypersensitivity disorder; AgNPs – silver nanoparticles; AuNPs –

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gold nanoparticles; BuP – butylparaben; BzP – benzylparaben; CFU – colony forming units;
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CIR – Cosmetic Ingredient Review; CuNPs – copper oxide nanoparticles; EtP –
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ethylparaben; EU – European Union; FDA – Food and Drug Administration; GMP – Good
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Manufacturing Practices; iso-BuP – iso-butylparaben; iso-PrP – iso-propylparaben; MCI –

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methylchloroisothiazolinone; MeP – methylparaben; MIT – methylisothiazolinone; PHBA –

4-parahydroxybenzoic acid; PrP – propylparaben; SCCS – Scientific Committee on Consumer

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Safety; TCS – triclosan; TiO2NPs – titanium dioxide nanoparticles; ZnONPs – zinc oxide
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nanoparticles

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 1. Introduction

Caring for personal hygiene is a basic way to prevent the occurrence and spread of

infectious diseases. Daily routines such as showering, brushing teeth, and washing face and

hair require the use of appropriate cosmetics: shampoos, toothpaste, and mouthwash. In

addition to personal care, cosmetics are used to enhance physical appearance and beauty, i.e.,

for making makeup and esthetic improvements.

The value of the global cosmetics market was estimated to be above 500 billion euros in

2018; moreover, this value is anticipated to increase further in coming years (Shahbandeh

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2019). Because of consumer demands for products with a long shelf life, the cosmetics

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industry uses several methods to prolong the period of use of cosmetics. One of the most
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common approaches used is the addition of preservatives, i.e., substances that inhibit the
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growth of microorganisms and counteract the generation of reactive oxygen species and
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oxidation processes in cosmetics. One of the most popular preservatives of cosmetics (and
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even pharmaceuticals and foodstuff) are parabens, which are esters of 4-parahydroxybenzoic

acid (PHBA). For the last 20 years, these compounds have been the topic of controversies
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among scientists and regular consumers. The debate is further stoked up by growing
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popularity of pro-ecologic and healthy lifestyle trends. It is thus no surprise that without a

clear indication of the negative effects of parabens on human health, cosmetic producers have

introduced alternative preservatives in the cosmetic market, which are advertised to be safer

than parabens (Błędzka et al. 2014; Nowak et al. 2018).

In the present review article, we have summarized literature data on the use of parabens

as a first-choice preservative in cosmetics and personal care products. We have summarized

the pros and cons of preservation by chemical and natural preservatives. We have reviewed

strategies for minimizing the use of parabens in cosmetic production and for preventing

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adverse health effects caused by parabens. Finally, we have considered the possibility of

absolute elimination of preservatives from personal care products and cosmetic ingredients.

2. Parabens

Parabens (esters of PHBA, nipagins, or aseptins) vary in their composition depending on the

type of substituents attached to PHBA. The simplest construction has methylparaben (MeP),

which is compound build on the PHBA framework with the methyl group as the substituent

[Figure 1]. By consecutively attaching methyl groups, a homologous series of 35 parabens

can be successively formed: ethylparaben (EtP), propylparaben (PrP), and butylparaben

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(BuP). Parabens with branched alkyl chains such as iso-propylparaben (iso-PrP) and iso-

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butylparaben (iso-BuP) as well as parabens with an aromatic ring such as benzylparaben
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(BzP) can also be produced from PHBA (Błędzka et al. 2014; Fransway et al. 2019;
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Matwiejczuk et al. 2020a; Nowak et al. 2018).
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The physicochemical properties of parabens [Table 1] favor their use as preservatives

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for various products. They are white, odorless crystals with a delicate taste and can dissolve in
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both water and organic solvents (the solubility in water decreases with the increasing length

of the chain of the substituent, while simultaneously increasing solubility in organic solvents).
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Their properties are stable and unaffected by changing temperature and physical state of the

preserved product. Moreover, the important advantage of parabens is that its mass production

is relatively simple and cheap (Fransway et al. 2019; Nowak et al. 2018).

The global popularity of parabens as preservatives is mainly because of their

effectiveness and activity against a broad spectrum of bacteria and fungi. They can inhibit the

growth of Gram positive (+) and Gram negative (–) bacteria, molds, and yeasts in preserved

products [Table 2] (Charnock and Finsrud 2007; Miceli et al. 2012; Murata et al. 2019;

Nemes et al. 2018). The antimicrobial properties of parabens increase with the increase in the

length of the alkyl chain in the substituent (MeP<EtP<PrP<BuP). These preservatives can

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inhibit DNA and RNA synthesis, inactivate bacterial enzymes, and affect lipid layers that lead

to disintegration of bacterial cell membranes (Flasiński et al. 2018; Nagar et al. 2019; Nes and

Eklund 1983; Shiralkar et al. 1978). A further characteristic that favors parabens as compared

to other preservatives is the activity of parabens in a broad range of pH. Several antimicrobial

substances such as sodium benzoate require acidic pH for their activity. On the other hand,

parabens may diffuse to oleic phase of emulsion, which requires the use of higher

concentration to attain effectiveness (Han and Washington 2005).

The maximal paraben content in cosmetics is regulated by the European Union,

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Scientific Committee on Consumer Safety (SCCS), and Food and Drug Administration (FDA)

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[Table 3]. It is mandatory for cosmetic producers to indicate the presence of parabens on the
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product package; moreover, in the case of paraben addition to foodstuff, it is necessary to use
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E symbols [Table 4]. Paraben usage at a concentration of lower than 0.4% for a single
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compound and 0.8% for mixture is regarded as safe (EC 1223/2009. 2009). However,
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according to Commission Regulation No. 1004/2014, the maximal concentration of PrP and

BuP cannot exceed 0.14% for a single compound and 0.8% for a mixture with other parabens.
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In contrast, SCCS recommends PrP and BuP as safe, if the concentration of either of them
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does not exceed 0.19%. Countries in the European Union have banned the use of long-chain

and branched parabens: iso-PrP, iso-BuP, BzP, phenylparaben, and pentylparaben (EC

358/2014. 2014). On the basis of the most recent scientific discoveries, the Cosmetic

Ingredient Review (CIR) Expert Panel in 2019 noted that the use of parabens as preservatives

should be repeatedly assessed. They highlighted that in addition to simple parabens, their

potassium and sodium salts are also used as cosmetic preservatives (CIR. 2019).

According to literature review, parabens are used in different types of personal care

products such as sanitary pads, panty liners, and tampons. Parabens are also added to

antibacterial creams, deodorants, and face powders. Among all parabens, MeP is the most

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commonly used one and in the highest concentration. Gao and Kannan (2020) showed that

MeP was present in 100% of the analyzed personal care products. MeP concentration is often

correlated with that of PrP. The content of other parabens in cosmetics decreases with an

increase in the chain length of the substituent. Branched parabens and those with aromatic

rings are rarely used and in very limited amounts (Błędzka et al. 2014).

Because parabens are ubiquitously used as cosmetic preservatives, the most common

way of exposition is its skin application. In research conducted on rabbits, scientists have

shown that MeP, EtP, and PrP penetrate skin independent of cream composition in line with

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decreasing solubility (MeP>EtP>PrP) (Pedersen et al. 2007). The process of skin diffusion of

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parabens may be enhanced by mechanical skin injury (Pažoureková et al. 2013). Scientists
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have observed that after dermal application, some part of parabens is accumulated (Pedersen
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et al. 2007), some part is hydrolyzed into PHBA (Jewell et al. 2007), and the remaining part in
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unchanged form may be systemically available (Pažoureková et al. 2013).

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It should be noted that paraben exposure is not only limited to dermal application, but it

may also be related to their inhalation or consumption; these exposure routes can cause
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extremely rapid absorption of parabens into the blood, lead to their ubiquitous distribution in
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the organism. The biotransformation processes mainly occur in microsomes of the small

intestine and liver, where human carboxylesterases (isoforms CES1 and CES2) exert their

enzymatic activity (Ozaki et al. 2013). Parabens are excreted in an unmetabolized form, as

conjugates with glucuronides and sulfates, PHBA, and p-hydroxyhippuric acid (Moos et al.

2016). These compounds have been identified in many biological materials, including serum,

urine, amniotic fluid, cord blood, milk, semen, adipose tissue, and hairs (Artacho-Cordón et

al. 2019; Buck Louis et al. 2018; Karzi et al. 2019; Li et al. 2020; Martins et al. 2020; Park et

al. 2019; Smarr et al. 2019; Song et al. 2020). Regardless of the analyzed material, the

percentage distribution of parabens reflects their content in the preserved products: MeP is the

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most frequently detected and in the highest concentration (often its concentration is correlated

with that of PrP). Other parabens commonly detected are EtP and BuP (Kolatorova et al.

2018a; Manouchehri et al. 2020; Wu et al. 2017).

Many myths, overinterpretations, and understatements have been made regarding the

adverse health effects of parabens. Before Darbre et al. (2004) published their study, in which

they suggested a relationship between the use of personal care products containing parabens

and its concentration in breast cancer tissue, these preservatives were recognised as safe.

Although the observations of Darbre et al. were never confirmed by other scientists, following

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this key publication, parabens started to be known as "cancer causing preservatives” (Flower

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2004; Harvey and Everett 2004; Jeffrey and Williams 2004).
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The most commonly occurring reaction to cosmetic compounds is contact sensitization;
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since the 90s this reaction is rarely noted for parabens and remains at stable level of 0.5-1%
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(Deza and Giménez-Arnau 2017; Giménez-Arnau et al. 2017). The biggest concern of
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paraben exposure is their ability to interfere with the functioning of the endocrine system; this

characteristic leads to their classification as endocrine disrupting chemicals. Parabens exhibit

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affinity (but lower than that of endogenic hormones) to receptors for estrogens, androgens,
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progesterone, thyroid hormones, peroxisome proliferator-activated receptor, and others from

the family of nuclear hormone receptors. Preservatives bind to receptors as their ligands and

affect the transcription of downregulated genes, which eventually may lead to disturbance of

body homeostasis (Fujino et al. 2020; Nowak et al. 2018; Nowak et al. 2019). It has been

experimentally confirmed that parabens may cause mitochondrial dysfunction and initiate

oxidative stress in cells (Kizhedath et al. 2019; Samarasinghe et al. 2018). Some scientists

have linked high level of paraben exposure with decreased quality of male semen, obesity

development, and immunological disorders (Jurewicz et al. 2017; Kolatorova et al. 2018b;

Nowak et al. 2020; Smarr et al. 2018). In vitro studies have revealed genotoxic and

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carcinogenic potential of parabens (Amin et al. 2019; Güzel Bayülken et al. 2018; Güzel

Bayülken et al. 2019; Güzel Bayülken and Ayaz Tüylü 2019; Lillo et al. 2017). The most

recent cohort research shed light on the relationship between fetus exposure to MeP during

pregnancy and the development of attention deficit hypersensitivity disorder (ADHD) in

childhood (Baker et al. 2020). Moreover, studies conducted on mice show that exposure to

parabens may cause effects on next generations of laboratory animals. On the other hand,

scientific literature also contains reports that paraben exposure does not cause negative

effects. In a study involved 215 young university male students Adoamnei et al. (2017) do

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not observe relationship between urinary concentration of parabens and decrease of semen

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quality parameters and derange reproductive hormone levels. Also, in a cross-sectional study
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with the participation of 4023 children form the U.S.A. researchers do not observe the
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associations between parabens exposure and asthma prevalence or asthma morbidity (Quirós-
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Alcalá et al. 2019).

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3. Alternative preservatives
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Because of a lack of clear statement regarding the toxicological profile of parabens, cosmetics

labelled as “PARABEN FREE” have gained increasing popularity. Moreover, the label
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“PARABEN FREE” is not officially registered in the European Union. Thus, cosmetics

producers are not legally obligated to fully eliminate parabens from the product ingredient

list. Consumers should also consider gains and losses of using products that are preserved by

“chemical” or “natural” paraben replacements. Some preservatives exhibit limited

antimicrobial activity, while others may act only in strictly defined conditions, while contact

with some of them may lead to similar or more harmful effects.

3.1.Chemical preservatives

One of the popular preservatives is formaldehyde, which is added to shampoos, soaps, nail

polishes, hair dyes, and conditioners [Figure 2]. To minimize the side effects of

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formaldehyde exposure, compounds that slowly release formaldehyde from their composition

are used in cosmetics, such as imidazolidinyl urea, diazolidinyl urea, DMDM hydantoin,

quaternium-15, and bronopol [Figure 1] (Fasth et al. 2018). The speed of formaldehyde

release depends on temperature, pH, time of storage, and product composition (Lv et al.

2015). Cosmetics containing more than 500 ppm of formaldehyde must be labelled as

“CONTAINS FORMALDEHYDE” (EC 1223/2009. 2009). Formaldehyde is considered as a

strong allergen, and contact with this preservative may induce chronic allergic eczema. It has

been observed that using skincare cosmetics containing a low amount of formaldehyde (2.5–

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40 ppm) in people with allergy led to dermatitis (Hauksson et al. 2016). In a 2010 published

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paper, the authors suggested a relationship between formaldehyde exposure and the
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development of acute myeloid leukemia (Zhang et al. 2010). However, their suspicions were
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never confirmed by other scientific groups (Allegra et al. 2019; Checkoway et al. 2015; Jones
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et al. 2015; Mundt et al. 2017; Speit et al. 2010).

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Triclosan (TCS) is a partially banned preservative in the European Union [Table 3].

TCS is used in personal care products such as toothpastes and mouthwashes, from where it
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absorbed in the organism. Within the first 4 days after exposure, approximately 24–83% of
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the absorbed TCS is excreted, while after 8 days of exposure, the level of excretion is close to

100% (Li et al. 2020; Lu et al. 2018; Sandborgh-Englund et al. 2006; Weatherly and Gosse

2017). Similar to parabens, TCS has been classified as an endocrine disrupting chemical.

According to scientific results, TCS affects the level of thyroid hormones, enhances oxidative

stress, increases the risk of asthma and allergy development, decreases the birth weight of

newborns, and negatively affects fertility (Clayton et al. 2011; Etzel et al. 2017; Koeppe et al.

2013; Lv et al. 2016; Spanier et al. 2014; Vélez et al. 2015). TCS shows low activity against

Gram-negative bacteria. Moreover, the use of TCS may indirectly result in the development

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of antibiotic resistance mechanisms in bacteria (Chen et al. 2009; Drury et al. 2013; Périamé

et al 2015).

Preservatives with effective antibacterial activity but also often cause allergies are

isothiazolinone releasers such as methylisothiazolinone (MIT), methylchloroisothiazolinone

(MCI), benzisothiazolinone, octylisothiazolinone, and dichlorooctylisothiazolinone. Because

of high risk of sensitization, MIT has been banned in leave-on products, while its maximal

concentration in rinse-off products (if present in a mixture with MCI) cannot exceed 0.0015%

[Table 3] (Aerts et al. 2014; Herman et al. 2019; Qin et al. 2019; Vanneste et al. 2013). A

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research study conducted in Spain showed the presence of isothiazolinones in 18% of

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cosmetics from local supermarkets, 7.9% of cosmetics from herbal shop, and 2.56% of

dermocosmetics (Pastor-Nieto et al. 2017).

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A common strategy of preservation of cosmetics and personal care products is the use of
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alcohols such as phenoxyethanol, benzyl alcohol, and their salts: sodium benzoate and
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potassium benzoate. The antibacterial activity of alcohols results from the inhibition of

enzyme synthesis and denaturation of cell membrane proteins (Lucchini et al. 1990).
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Phenoxyethanol, benzyl alcohol, and their salts are absorbed into the body through the
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gastrointestinal system and transdermally. Phenoxyethanol is oxidized and excreted as 2‐

phenoxyacetic acid through urine. Benzyl alcohol is metabolized to benzoic acid, which (like

sodium benzoate) is linked to glycine and excreted as hippuric acid through urine (Nair 2001).

The CIR Expert Panel recognized that benzyl alcohol and its derivatives exhibit low general

toxicity and rarely irritate skin (Johanson et al. 2017). Similarly, Dréno et al. (2019) based on

the review of available literature conclude that phenoxyethanol should be recognized as safe if

used as a cosmetic preservative in concentration not exceeding 1%. Benzyl alcohol and

sodium benzoate are used in toothpastes, mouthwashes, facial cleansers, eye makeup

cleansers, and face and hand creams (Baranowska et al. 2014). According to Panico et al.

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(2019), cosmetics available in Italy are most frequently preserved by phenoxyethanol

(47.8%), sodium benzoate (35.6%), and potassium sorbate (22%) [Figure 2].

Some of the preservatives such as formic acid, sorbic acid, propionic acid, citric acid,

and benzyl alcohol are present in nature. However, when used in cosmetics, these compounds

are not purified products of natural origin but are chemically synthesized in factories (Ricke

2003). This strategy is used by dishonest cosmetic producers who falsely advertise their

products as “completely natural.”

3.2.Natural preservatives

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Recently, the cosmetics industry started a new trend of replacement of chemical preservatives

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with substances of only natural origin. Scientists have identified plants that exhibit
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antibacterial and antifungal properties and have assessed the effectiveness, efficiency, and
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workable possibilities of using essential oils and extracts derived from these plants in
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cosmetics (Herman 2019; Pandey et al. 2017; Xiao et al. 2020). Since ages, people are aware
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of the antimicrobial properties of plants such as garlic (Allium sativum), thyme (Thymus
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vulgaris), cinnamon (Cinnamomun), tea (Camellia sinensis), and turmeric (Curcuma longa)

(Marchese et al. 2014; Nabavi et al. 2015). Many plant metabolites such as lectins, alkaloids,
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terpenoids, polypeptides, polyphenols, and polyacetylenes show antibacterial and antifungal

activity. Furthermore, plant derivatives are used in food industry and many of them are

classified as “GRAS” – Generally Recognized as Safe (Simões et al. 2009). Thus, these

substances are now being commonly used in cosmetics.

A recent study demonstrated that extracts of Lonicera japonica and Magnolia obovata

effectively inhibit the growth of Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus

subtilis, and Candida albicans (Lee et al. 2018). Tea (C. sinensis) extract has a beneficial

effect on skin microcirculation, improves hair condition, and exhibits anticellulite,

antioxidant, photoprotective, and slimming properties. Moreover, it does not cause skin

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irritation and allergies and thus has found broad application in cosmetics industry. Perhaps,

because of its antibacterial potential, tea extracts will also be used as preservatives in the

future (Farooqui et al. 2015; Koch et al. 2019). Cinnamon (Cinnamomum) extracts and oils

are well known for their antibacterial and antifungal properties; these compounds inhibit

Bacillus licheniformis, Bacillus megaterium, Corynebacterium xerosis, Escherichia coli,

Enterobacter cloacae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Pseudomonas

fluorescens, Salmonella typhi, Salmonella paratyphi A, Staphylococcus aureus, Streptococcus

faecalis, Yersinia enterocolitica, and other bacterial species (Keskin and Toroglu 2011;

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Nabaviet al. 2015; Naveed et al. 2013). A new hope for fans of natural conservation are the

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results of studies conducted by Herman (2014) and Herman et al. (2013). This research team
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compared the antibacterial and antifungal activity of MeP, plant extracts (Matricaria
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chamomilla, Aloe vera, and Calendula officinalis), and essential oils (Lavandulla officinalis,
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Melaleuca alternifolia, and Cinnamomum zeylanicum). The most effective compound in

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terms of growth inhibition of bacteria, yeasts, and molds was cinnamon oil (the observed

effect was stronger than that observed for MeP). Based on the observed results, the authors
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recommended the use of cinnamon oil instead of traditional cosmetic preservatives.

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Particularly promising results of inhibition of mycelial growth and germination of spores of

Aspergillus niger were obtained using nanoemulsion from essential oil of cinnamon leaves.

Ribes et al. (2017) believed that their results will help to discover efficient and stable natural

antifungal preservatives based on the nanoemulsion technique.

Some plant-derived antimicrobial substances are already commercially used in

cosmetics. CIR reported that extracts from fruits and leafs of Carica papaya are used in

lipsticks, face powders, pump spray suntan products and deodorants (CIR. 2020). Moreover,

Saliasi et al. (2018) show C. papaya leafs extracts reduces interdental bleeding thus it could

be a precious compound of toothpaste or mouthwash. So far, C. papaya-derived ingredients

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are not restricted in any way from use in cosmetic products in the European Union (CosIng

database. 2009). Because of antimicrobial properties, also citrus extracts are used in products

available in the market for maintaining oral hygiene (Cieplik et al. 2019). An example of the

natural-origin antibacterial and antifungal product for maintenance mouth hygiene is Citrix®,

whose components are based on soluble bioflavonoids derived from citrus fruits (Hooper et

al. 2011). Besides to ability to reduce the nasopharyngeal microbiota, the citric extract may

also reduce viral load. Thus Carrouel et al. (2020) suggested that citric-based mouth hygiene

product should be considered as an adjunct in prevention against SARS-CoV-2 infection.

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Essential oils from thyme (T. vulgaris) and carnation (Syzgizum aromaticum), whose

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main active metabolites are thymol and eugenol, exhibit numerous benefits. Oils from these
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plants are recognized as antioxidants; exhibit anti-inflammatory, neuroprotective, and
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cardioprotective properties, and act against Gram-positive and Gram-negative bacteria and
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fungi. Given these properties, thymol and eugenol are used in food production. In recent past,
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scientists have conducted first attempts to use them in cosmetics as natural competitors of

chemical preservatives (Orchard and van Vuuren 2017; Salehi et al. 2018). Oils from tea tree
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(Melaleuca alternifolia), lavender (Lavandula luisieri), and rosemary (Rosmarinus officinalis)

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are free radical scavengers with antibacterial and antifungal properties (Giménez-Rota et al.

2019; Rašković et al. 2014; Zhang et al. 2018; Zuzarte et al. 2012). Presently, essential oils

are being increasingly shown to be effective and promising antifungal substances. Because

essential oils contain a mixture of different active components without specific modes of

action, fungi cannot form adaptive mechanisms and resistance (Carson et al. 2002).

The complete elimination of chemical preservatives from cosmetics and their

replacement with natural substances may be problematic. The main challenge for producers of

cosmetics preserved with only natural compounds is to maintain the shelf life at a level

similar to that of chemically preserved products. It has been shown that thymol exhibits

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antimicrobial properties against E. coli, S. aureus, and P. aeruginosa maximally for 2 weeks.

Encapsulated thymol is more efficient and may protect the product against bacterial growth

for 3 months. Surprisingly, encapsulated thymol exhibits antimicrobial properties similar to

those of MeP, even when used at much lower concentrations (12–52 times) (Wattanasatcha et

al. 2012). Although the use of nano-encapsulation may increase the bioactivity of the active

substance of cosmetics and simultaneously decreases its amount, the overall costs of materials

and manufacture very often exceed the cost of traditional and well-known methods of

preservation. Thus, the production of nanoparticle-based preservative may generate additional

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costs, which will increase the final cost of the product. (Carrouel et al. 2020). Other important

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challenges related to the use of plant oils as preservatives are low solubility, strong smell, and
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frequent interactions with other cosmetic ingredients (Halla et al. 2018; Samperio et al. 2010).
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Mantil et al. (2015) highlighted an obstacle for the common use of essential oil of tea tree
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(Melaleuca alternifolia) as a preservative. This substance shows low activity in the water
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phase of cosmetic, but as a solution to this problem, scientists suggest an increase in lipid

phase contribution in cosmetic formula. Finally, the biggest weakness of natural preservatives
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– organicity – is regarded as their biggest strength. Natural compounds are rich in macro- and
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microelements, which are an excellent source of nutrients for microorganisms, against which

preservatives should protect (Narayanan et al. 2016).

4. Strategies to limit the use of paraben

The preservation of cosmetic products by using only one preservative is rare. More

frequently, cosmetic producers formulate a mixture of preservatives from different groups.

This formulation is based on the natural properties of the included preservatives: some of

them can act only against Gram-positive bacteria, while others cannot inhibit fungi. Mixtures

of preservatives allow to maintain a broad spectrum of antimicrobial protection and enable to

minimize the content of particular preservatives (Smaoui and Hlima 2012). One of the ideas

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for minimizing the content of long-chain parabens and simultaneously provide efficient

protection against the development of microorganisms is to prepare a mixture of different

parabens at the lowest possible concentration (Charnock and Finsrud 2007).

In addition to compounds officially classified as preservatives, cosmetics contain

multifunctional substances, including a preservative booster. Mixing preservatives and

boosters enables to decrease the concentration of added parabens; more importantly, this

procedure does not decrease their preservative potential but may minimize paraben-caused

adverse effects. Multifunctional substances approved for use in cosmetics are glycols, fatty

of
acids and their monoesters, phenethyl alcohol, ethylhexylglycerin, and phospholipids

ro
(Narayanan et al. 2016). When used alone, these compounds cannot efficiently preserve, e.g.,
-p
ethylhexylglycerin inhibits the growth of Gram-positive bacteria but does not affect Gram-
re
negative bacteria and fungi. However, it is used in the cosmetics industry because of its other
lP

properties such as moisturizing, oiling, surface-activeness, antioxidant, or weak bactericidal

na

activity (Herman 2019). Compounds from natural origin could also be used as preservatives

boosters. According to the most recent research, mixing parabens with sulforaphane
ur

(isothiocyanate, found in cruciferous vegetables, e.g., broccoli, cauliflower, brussels sprouts,

Jo

cabbage, and arugula) enhances its antifungal activity by facilitating the penetration of the cell

membrane of microorganisms (Murata et al. 2019). Surfactants and other active substances

from essential oils disintegrate bacteria cell membranes, thus sensitizing bacteria for

penetration by the antimicrobial compounds. A synergistic effect is observed by mixing

essential oils from marjoram (Origanum majorana), eucalyptus (Eucalyptus globosus), mint

(Mentha piperita), clove (Salvia officinalis), and rosemary (Rosmarinus officinalis) and

surfactants with preservatives enable to decrease the concentration of parabens without

decreasing its antimicrobial potency (Patrone et al. 2010).

17
 Another strategy to minimize paraben-induced adverse health effect is the addition of

other substances that will avoid the negative effect of parabens in the cosmetic formulation.

Majewska et al. (2017) showed the negative effect of MeP on human fibroblasts (enhanced

apoptosis, inhibition of collagen synthesis and cell proliferation). In another study, researchers

demonstrated the protective effect of rosemary acid on fibroblasts exposed to MeP

(Matwiejczuk et al. 2020b). Although these studies were conducted on in vitro cell culture

models, the obtained results are promising; thus, the efficiency of the proposed solution

should be validated by using different methodologies, including the use of model with

of
laboratory animals or system of skin EpiSkin™ (Deshmukh et al. 2012).

ro
Perhaps, in the near future, scientists will develop new methods of preservation, which
-p
will completely replace “chemical” preservatives. The most recent trend introduced in the
re
market of cosmetic preservatives is nanoparticles. Nanoparticles of zinc oxide (ZnONPs),
lP

silver (AgNPs), gold (AuNPs), titanium dioxide (TiO2NPs), and copper oxide (CuNPs) are
na

used in toothpastes and mouthwashes. ZnONPs generate reactive oxygen species and release

cytotoxic Zn2+ ions, which lead to the disintegration of the cell membrane of Gram-positive
ur

and Gram-negative bacteria. AgNPs exert antibacterial activity in both oil and water phase of
Jo

a preserved product (Carrouel et al. 2020; Hosny et al. 2017; Kokura et al 2010; Salvioni et al.

2017). As nanoparticles are cutting-edge technology, this method of preservation has raised

many questions about their long-term effect on humans.

5. World without preservatives?

The most common microorganisms that contaminate cosmetics are bacteria, including P.

aeruginosa, Burkholderia cepacia, Klebsiella oxytoca, E. coli, S. aureus, Serratia

marcescens, and Enterobacter gergoviae, yeasts such as Candida albicans, and molds. The

abovementioned species are potential pathogens and may cause adverse health effects.

Additionally, exposure to microbial-contaminated products may challenge the immune

18
system, which is a natural barrier against microbial invasion and their potential negative

effects on the organism (SCCS/1564/15. 2016).

Cosmetics and personal care products available in the market are not sterile. These

products contain a low amount of microorganisms, which when present at low concentration

does not pose a serious threat to humans. The regulation of the European Union states that

microbial contamination in cosmetics and personal care products cannot exceed:

a. 100 colony forming units (CFU)/g or 100 CFU/mL in products used in eyes and

mucosal area for children under 3 years of age,

of
b. 100 CFU/g or 1000 CFU/mL in other products.

ro
Moreover, cosmetic producers should monitor the presence of bacteria with greatest
-p
pathogenic potential, such as P. aeruginosa, S. aureus, and C. albicans (SCCS/1564/15.
re
2016). In the United States, cosmetic products can be launched in the market, in which the
lP

level of microorganisms does not exceed:

na

a. 500 CFU/g in products used in eye area and those dedicated to infants,

b. 1000 CFU/g in other products (Huang et al. 2017).

ur

To meet the abovementioned restrictions, cosmetics undergo two-steps preservation: primary

Jo

and secondary (addition of preservatives into the manufactured product). In response to

controversies regarding some preservatives, primary preservation implies procedures

conducted in the phase of cosmetic preparation and manufacture. According to the Cosmetics

Regulation EC 1223/2009 (2009), the maintenance of “Good Manufacturing Practices”

(GMP) (ISO22716 regulation) is required for cosmetics distribution in the European cosmetic

market. GMP sets guidelines that minimize the possibility of microbial over-contamination in

the process of cosmetic production [Figure 3]. According to these guidelines, the production

line staff should be trained properly, the production equipment used should be regularly

19
disinfected and calibrated, and the production area should be cleaned regularly to ensure

maximum product protection (EC 1223/2009. 2009; GMP. 2016).

Based on debate around safety of preservatives raised idea of self-preserving cosmetics.

In the phase of product formulation and manufacturing process, the cosmetics producers

implement strategies that allow to minimize the amount of added preservatives. One of the

proposed strategies is to limit the water content in cosmetics as it is the preferred environment

for growth by bacteria and fungi; this step prevents the development of microorganisms.

Formulations based on water-in-oil (W/O) emulsions are less susceptible to microbial growth

of
than oil-in-water (O/W) emulsions. Moreover, the development of microorganisms is less

ro
common in oil phase-based nanoemulsions (however, their antibacterial activity depends on
-p
composition) (Buranasuksombat et al. 2011; Char et al. 2016; Salvia-Trujillo et al. 2015;
re
Varvaresou et al. 2009).
lP

The growth rate of bacteria and fungi is reduced in cosmetics with pH exceeding the
na

preferable range (pH: 5–8). The acidic pH of antiperspirants is ensured by the addition of

ascorbic acid or aluminum compounds, while alkaline pH is common in liquid soaps.

ur

However, ensuring a reduced or elevated pH is problematic for loose products and may cause
Jo

skin irritation (Halla et al. 2018; Varvaresou et al. 2009).

Finally, the method of packaging of finished products is also important. Vacuum

packaging is used to minimize the exposure of cosmetics to air (which is a medium for

microorganisms). Enclosing products in screw-capped jars and containers is not a sufficient

barrier to microorganisms. A much more effective, though not perfect, approach is to use pipe

systems ending with valves or equipped with pumps. Disposable containers, sachets, and

capsules are also available; this type of product packaging, although effectively protects

against microbial contamination, generates additional costs and extensive waste (Halla et al.,

2018; Varvaresou et al. 2009).

20
 6. Conclusion

One of the most commonly used preservatives in cosmetics and personal care products are

parabens. Because these compounds are absorbed into human body directly from the

preserved products and cause adverse health effects, some of the regulatory agencies have

restricted the use of parabens in cosmetics. To ensure an efficient control of microbial over-

contamination, minimize paraben-caused adverse health effects, and simultaneously avoid

exceeding the maximum allowable doses of parabens, the following strategies are being

implemented: (1) replacement of parabens with other chemical or natural preservatives, (2)

of
formulation of mixtures of parabens with other preservatives, (3) combination of parabens

ro
with preservative boosters, (4) preparation of composite mixtures of various parabens in ideal
-p
proportions to ensure low concentrations, and (5) following GMP in cosmetics, particularly in
re
the primary maintenance stage.
lP

Consumer interest, in terms of effectiveness and more importantly safety of the

na

cosmetic products available in the market, is an important factor for developing the best

product preservation strategies. Regardless of whether preservatives will have a “chemical” or

ur

“natural” origin, the key to create a perfect cosmetic seems to be finding the perfect
Jo

proportions of the ingredients that constitute it. On the basis of the data available in the

literature, it is now possible to use strategies to prevent the negative effects of preservatives

while providing effective protection of cosmetics against the growth of microorganisms.

7. Acknowledge

This research did not receive any specific grant from funding agencies in the public,

commercial, or not-for-profit sectors.

8. Declaration of interest statement

Authors declare no commercial or financial conflict of interest.

21
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10. Tables

Table 1. Characteristic of chemical preservatives used in cosmetics and presonal care

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products. Chemical preservatives (methylparaben, ethylparaben, propylparaben, butylparaben,

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methylisothiazolinone, methylchloroisothiazolinone, triclosan, phenoxyethanol, sodium
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benzoate, benzyl alcohol and potassium sorbate) were compared in regard to its structure and
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physicochemical properties (Pubchem: 7456, 8434, 7175, 7184, 39800, 33344, 5564, 31236,
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517055, 244, 236745).

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Table 2. Antimicrobal activity of parabens. Esters of p-hydroxybenzoic acid are able to kills
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bacteria Gram positive, Gram negative and fungi.

Table 3. Legal regulations in the European Union of preservatives used in cosmetics and
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personal care products.

Table 4. E symbols of food preservatives.

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 11. Figures

Figure 1. Structure of compounds commonly used in cosmetics as preservatives. Numerous

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compounds exhibit antimicrobial properties, some of these (e.g. alcohols, aldehydes and

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formaldehyde-releasers, isothiazolinones, organic acids and phenols (parabens and triclosan))

are used in cosmetics preservation.

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Figure 2. Preservatives declared on cosmetics labels. Data express percentage of cosmetics
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from different countries (Spain, Italy, Sweden, Israel, Denmark, Thailand, Germany,
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Switzerland, U.S.A.) which contains preservatives (parabens, formaldehyde releasers,

methylchloroisothiazolinone and/or methylisothiazolinone, phenoxyethanol, sodium benzoate,

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potassium sorbate and triclosan) according to information from labels. Legend: Country (N of
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tested cosmetics) (Rastogi 2000, De Groot and Venstra 2010, Yazar et al. 2011, Uter et al.

2014, Horev et al. 2015, Bunyavaree et al. 2016, Garcia-Hidalgo el al. 2017, Pastor-Nieto et

al. 2017. Panico et al. 2019)

Figure 3. Good Manufacturing Practice in Cosmetics. According to Good Manufacturing

Practice (GMP) for cosmetic products: ISO 22716, production of cosmetics must follow

through rules and procedures which ensure high quality of final product. Each step of

production should be documented and controlled. Only qualified staff could work on well-

adjusted premises and use validated equipment. Final product must be manufactured in

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accordance with established procedures regarding handling of deviations, waste, packaging

and storage.

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Compoun Methylpara Ethylparabe Propylpara Butylparab Methylisothiazoli Methylchloroisothiazolin Triclosan Phenoxyetha Sodium Benzyl Potassiu
d ben n ben en none one nol benzoate alcohol m
sorbate
Molecula C8H8O3 C9H10O3 C10H12O3 C11H14O3 C4H5NOS C4H4ClNOS C12H7Cl3 C8H10O2 or C7H5O2Na C7H8O or C6H7O2K
r formula O2 C6H5OC2H4 or C6H5CH2 or
OH C7H5NaO2 OH C6H7KO2
or
C6H5COO
Na or
NaC6H5C
OO
Molecula 152.15 166.17 180.2 g/mol 194.23 115.16 g/mol 149.6 g/mol 289.5 138.16 g/mol 144.1 108.14 150.22

f
r weight g/mol g/mol g/mol g/mol g/mol g/mol g/mol

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CAS no. 99-76-3 120-47-8 94-13-3 94-26-8 2682-20-4 26172-55-4 3380-34- 122-99-6 532-32-1 100-51-6 590-00-1
5

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European 243-171-5 204-399-4 202-307-7 202-318-7 220-239-6 247-500-7 222-182- 204-589-7 208-534-8 202-859-9 246-376-

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Commun 2 1
ity

Pr
number
Color white white white white colorless - white colorless white colourles white

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Odor odorless odorless odorless odorless - - slight, faint almost faint character
faintly aromatic odourless aromatic istic

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aromatic
Taste slight slight slight slight
u - - - burning sweetish, sharp -
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burning burning burning burning astringent burning
Form cristals cristals cristals cristals liquid cristals cristals liquid crystalline liquid powder
powder or or
granules crystals
Boiling 270.5°C 297.5°C 271°F at 1 - - - 120°C 245°C >450°C to 205.3°C -
point mm Hg <475°C
Melting 131°C 117.0°C 203 to 154 to 50-51°C 54-55°C 55-57°C 14°C 436°C -15.2°C >270°C
point 208°F 156°F
Table 1. Characteristic of chemical preservatives used in cosmetics and presonal care products. Chemical preservatives (methylparaben,

ethylparaben, propylparaben, butylparaben, methylisothiazolinone, methylchloroisothiazolinone, triclosan, phenoxyethanol, sodium benzoate,

benzyl alcohol and potassium sorbate) were compared in regard to its structure and physicochemical properties (Pubchem: 7456, 8434, 7175,

7184, 39800, 33344, 5564, 31236, 517055, 244, 236745).

Parabens antimicrobal activity
Bacteria Gram positive Lactobacillus
Streptococcus mutans
Staphylococcus aureus
Staphylococcus aureus
Micrococcus luteus
Staphylococcus aureus
Staphylococcus aureus
Staphylococcus aureus
Staphylococcus aureus
Bacteria Gram negative Escherichia coli
Escherichia coli
Pseudomonas aeruginosa
References
Steinberg et al. 1999
Charnock and Finsrud 2007
Młynarczyk et al. 2008
Smaoui and Hlima 2012
Crovetto et al. 2017
Flasiński et al. 2018
Nemes et al. 2018
Bredin et al. 2005
Nguyen et al. 2005
Charnock and Finsrud 2007

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Escherichia coli Młynarczyk et al. 2008
Pseudomonas aeruginosa

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Salmonella Gurtler and Jin 2012
Escherichia coli Smaoui and Hlima 2012
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Pseudomonas aeruginosa
Salmonella
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Escherichia coli Crovetto et al. 2017
Pseudomonas aeruginosa Flasiński et al. 2018
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Escherichia coli Nemes et al. 2018

Pseudomonas aeruginosa
Escherichia coli Murata et al. 2019
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Fungi Candida albicans Charnock and Finsrud 2007

Zygosaccharomyces bailii,
Aspergillus niger Młynarczyk et al. 2008
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Candida albicans
Candida albicans Miceli et al. 2012
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Aspergillus niger Smaoui and Hlima 2012

Candida albicans
Candida albicans Flasiński et al. 2018
Candida albicans Nemes et al. 2018
Candida glabrata
Candida parapsilosis
Aspergillus niger Murata et al. 2019
Candida albicans
Saccharomyces cerevisiae

Table 2. Antimicrobal activity of parabens. Esters of p-hydroxybenzoic acid are able to kills

bacteria Gram positive, Gram negative and fungi.

Compound Legal regulations References

Parabens maximum concentration limits of methylparaben or ethylparaben (single esters and their salts) is sets as 0.4% EC 1223/2009. 2009

maximum concentration limits of methylparaben or ethylparaben (single esters and their salts) is sets as 0.4% SCCS/1514/13. 2013

maximum concentration limits of methylparaben and ethylparaben in mixture is sets as 0.8% SCCS/1514/13. 2013

maximum concentration limits of propylparaben and butylparaben in mixture is sets as 0.19% SCCS/1514/13. 2013

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maximum concentration limits of butylparaben or propylparaben (single esters and their salts) is sets as 0.14% EC 1004/2014. 2014

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maximum concentration limits of mixture of methylparaben, ethylparaben, butylparaben and propylparaben is EC 1004/2014. 2014

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sets as 0.8% (while sum of the individual concentration of butylparaben and propylparaben cannot exceed

0.14%)

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iso-propylparaben, iso-butylparaben, phenylparaben, benzylparaben, and pentylparaben are not approved as EC 358/2014. 2014

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preservatives in cosmetic products

Methylisothiazolinone not approved in leave-on products EC 1223/2009. 2009

maximum concentration in rinse-off products is set as 0,0015% (15 ppm) SCCS/1557/15. 2015

EC 2017/1224. 2017

Triclosan not approved in biocidal products for hygiene purposes (for example: hand disinfectants) EC 528/2012. 2012

not approved as an active substance for use in biocidal products for product-type 1 EC 2016/110. 2016
 maximum concentration in toothpastes, hand soaps, body soaps, shower gels, deodorants (non-spray), face EC 358/2014. 2014

powders, blemish concealers, nail products for cleaning the fingernails and toenails before the application of

artificial nail systems is sets as 0.3%

maximum concentration in mouthwashes is set as 0.2% EC 358/2014. 2014

Phenoxyethanol maximum concentration is sets as 1.0% EC 1223/2009. 2009

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Sodium benzoate maximum concentration in rinse-off products, except oral products is set as 2,5% (acid) EC 1223/2009. 2009

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maximum concentration in oral products is set as 1,7% (acid) EC 1223/2009. 2009

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maximum concentration in leave-on products is set as 0,5% (acid) EC 1223/2009. 2009

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Potassium sorbate maximum concentration in cosmetics products is sets as 0,5% (acid) EC 1223/2009. 2009

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Table 3. Legal regulations in the European Union of preservatives used in cosmetics and personal care products.
E symbol Compound
E200 Sorbic acid
E201 Sodium sorbate
E202 Potassium sorbate
E203 Calcium sorbate
E209 Heptyl p-hydroxybenzoate
E210 Benzoic acid
E211 Sodium benzoate
E212 Potassium benzoate
E213 Calcium benzoate
E214 Ethylparaben
E215 Sodium ethyl para-hydroxybenzoate
E216 Propylparaben
E217 Sodium propyl para-hydroxybenzoate
E218 Methylparaben

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E219 Sodium methyl para-hydroxybenzoate
E220 Sulfur dioxide

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E221 Sodium sulfite
E222 Sodium bisulfite
E223
E224
Sodium metabisulfite
Potassium metabisulfite
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E225 Potassium sulfite
E226 Calcium sulfite
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E228 Potassium hydrogen sulfite

E234 Nisin
E235 Natamycin, Pimaracin
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E236 Formic acid

E237 Sodium formate
E238 Calcium formate
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E239 Hexamine
E240 Formaldehyde
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E242 Dimethyl dicarbonate

E249 Potassium nitrite
E250 Sodium nitrite
E251 Sodium nitrate
E252 Potassium nitrate
E262 Sodium acetates, Sodium acetate, Sodium diacetate
E264 Ammonium acetate
E265 Dehydroacetic acid
E266 Sodium dehydroacetate
E280 Propionic acid
E281 Sodium propionate
E282 Calcium propionate
E283 Potassium propionate
E284 Boric acid
E285 Sodium tetraborate
Table 4. E symbols of food preservatives.
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Declaration of interests

☒ The authors declare that they have no known competing financial interests or personal relationships
that could have appeared to influence the work reported in this paper.

☐The authors declare the following financial interests/personal relationships which may be considered
as potential competing interests:

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