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Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
1
Hidratante labial usando oleoresina e óleo essencial de canela: avaliação da segurança
microbiológica e estabilidade estendida e prolongada
Lip balm using cinnamon oleoresin and essential oil: microbiological safety assessment
with accelerated and extended stability
Bálsamo labial con oleorresina y aceite esencial de canela: evaluación de seguridad
microbiológica y estabilidad acelerada y prolongada
Received: 12/08/2020 | Reviewed: 23/08/2020 | Accept: 25/08/2020 | Published: 29/08/2020
Leonam José de Andrade Lima
ORCID: https://orcid.org/0000-0002-0067-0819
Federal University of Triângulo Mineiro, Brazil
E-mail: leonamjose.andradelima@gmail.com
Luana Cominato
ORCID: https://orcid.org/0000-0002-1633-887X
Federal University of Triângulo Mineiro, Brazil
E-mail: luanacominato@hotmail.com
Heloísa Moraes Oliveira
ORCID: https://orcid.org/000-0002-6594-7612
Federal University of Triângulo Mineiro, Brazil
E-mail: heloisamoli@outlook.com
Wanderson de Oliveira dos Santos
ORCID: https://orcid.org/0000-0002-8292-608X
Ourofino Agrociências, Brazil
E-mail: wanderson.santos@ourofinoagro.com.br
Geoffroy Roger Pointer Malpass
ORCID: https://orcid.org/0000-0002-0036-5750
Federal University of Triângulo Mineiro, Brazil
E-mail: geoffroy.malpass@uftm.edu.br
Monica Hitomi Okura
ORCID: https://orcid.org/0000-0002-9875-9378
Federal University of Triângulo Mineiro, Brazil
E-mail: monica.okura@uftm.edu.br
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
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Ana Claudia Granato
ORCID: https://orcid.org/0000-0001-6487-1225
Federal University of Triângulo Mineiro, Brazil
E-mail: ana.malpass@uftm.edu.br
Resumo
Devido ao crescimento da indústria cosmética, são necessários desenvolvimentos e melhorias
para o setor produtivo e seus produtos, para que atendam à demanda de consumo e também
aos padrões de qualidade impostos pelos órgãos de inspeção. Em vista disso, um ramo que se
desenvolveu para atender a essas necessidades é o dos fitocosméticos, que usam nos
constituintes de suas formulações de origem vegetal com atividades biológicas. O objetivo
deste trabalho foi comparar as formulações de hidratantes labiais incoporando-se oleorresina e
óleo essencial de canela (Cinnamomun verum), além de testar a segurança microbiológica e a
estabilidade acelerada e estendida dos produtos formulados. Os testes revelaram que as
formulações propostas são estáveis e seguras microbiologicamente, o que se deve à atividade
antimicrobiana da oleorresina e óleo essencial de canela utilizados nas formulações.
Palavras-chave: Hidratante labial; Canela; Oleorresina; Óleo essencial.
Abstract
Due to the growth of the cosmetics industry, developments and improvements are needed for
the productive sector and its products to meet consumer demand and the quality standards
imposed by inspection bodies. As a result, an area has developed to meet these needs that is
known as phytocosmetics. This area involves the inclusion of constituents with of plant origin
in the cosmetic product formulation. The objective of this work was to compare the
formulations of lip moisturizers incorporating oleoresins and essential oils of cinnamon
(Cinnamomun verum), in addition to testing the microbiological safety and the Accelerated
and Extended Stability of the formulated products. The tests revealed that the proposed
formulations are microbiologically stable and safe, which is due to the antimicrobial activity
of the oleoresin and cinnamon essential oil used in the formulations.
Keywords: Lip balm; Cinnamon; Oleoresin; Essential oil.
Resumen
Debido al crecimiento de la industria cosmética, se necesitan desarrollos y mejoras para el
sector productivo y sus productos, a fin de satisfacer la demanda del consumidor y también
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
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los estándares de calidad impuestos por los organismos de inspección. En vista de esto, una
rama que se ha desarrollado para satisfacer estas necesidades es la de los fitocosméticos, que
utilizan en los componentes de sus formulaciones de origen vegetal con actividades
microbiológicas. El objetivo de este trabajo fue comparar las formulaciones de humectantes
labiales que incorporan oleorresinas y aceites esenciales de canela (Cinnamomun verum),
además de probar la seguridad microbiológica y la estabilidad acelerada y extendida de los
productos formulados. Las pruebas revelaron que las formulaciones propuestas son
microbiológicamente estables y seguras, lo que se debe a la actividad antimicrobiana de la
oleorresina y el aceite esencial de canela utilizados en las formulaciones.
Palabras clave: Bálsamo labial; Canela; Oleorresina; Aceite esencial.
1.Introduction
A cosmetic is defined as any substance or preparation intended to be placed in contact
with the various external parts of the human body (epidermis, capillary system, nails, lips and
external genital organs) or with the teeth and mucous membranes of the oral cavity, with the
purpose of modifying their appearance, correcting body odour and/or protecting them by
keeping them in good condition. Cosmeceuticals are intended to perform their functions, such
as protection, whitening, tanning, anti-wrinkle, deodorants, anti-aging and nails and hair care
as a cosmetic product, but also claim to have biologically active ingredients with medicinal
benefits (Aranaz et al., 2018).
Products with topical action should be formulated to ensure that the active compounds
efficiently cross the different layers of tissue to maximize their effectiveness. Although the
skin constitutes a multi-layered organ, it is the outermost layer, which exerts the greatest
barrier action to the permeation of drugs (Lane, 2013). The lips are an extremely sensitive
region and have skin three times thinner than the other regions of the body. The lips do not
have follicles, do not produce sebaceous secretions and are not covered, unlike other regions
of the body, by the protective lipid film. Therefore, lips are very prone to dehydration and
cracking. In addition, as saliva is constantly secreted from within the mouth and has saline
properties, cosmetic products for this region require the pH must be compatible with the pH
of human saliva (Galembeck & Csordas, 2009).
Lip balm is a cosmetic product similar to lipstick and is employed to avoid lip dryness
and provide protection against adverse environmental factors. The product in question should
possess the following characteristics: pleasant taste, resistance to temperature variations,
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
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softness in the application, easy intentional removal and adherence (Fernandes et al., 2013).
In addition to the use of waxes, oils and butters, which are responsible for providing
consistency, antioxidants, preservatives, fragrances, dyes and pigments can be used in lip
balm formulations (Denavarre, 1975; Barel, Paye & Maibach, 2009).
Since ancient times, natural resources are well known as a source of biologically
active substance to be used as cosmetics or cosmeceutical products (Aranaz et al, 2018).
Among them are essential oils and oleoresins, which are natural products of the secondary
metabolism of plants. Essential oils and oleoresins are known to present a broad spectrum of
action, including antioxidant activity and antibacterial activity. Even though the antimicrobial
mechanism of action of essential oils and oleoresins is not fully understood, it is believed that
the activity occurs due to their hydrophobicity (Martinelli et al., 2017; Rosa et al, 2018).
Essential oils have a recognized aesthetic and therapeutic potential and are mainly
used for their bactericidal, virucide, antioxidant and anti-inflammatory activities, but also for
their fragrant character, which can cause a sense of well-being. Several ways to use essential
oils are possible: inhalation, ingestion, or application to the skin. Pure essential oils are almost
never applied directly to the skin, as they can be irritating, but are diluted in other vegetable
oils (olive oil, sunflower seed oil, etc.). The application to the skin is performed in the context
of massages, local treatments (infections) or when using perfumes (main constituents) (Marrot
& Soeur, 2015).
According to Eiserle & Rogers (1972), essential oils and oleoresins are especially
important flavour constituents for hot processed foods. However, according to the authors,
essential oils tend to volatilize during processing due to the heat and humidity present, losing
some of the flavouring components. However, as they contain essential oils and fixatives
compounds, oleoresins tend to depress volatilization and are preferred as flavouring materials.
This is because solvent extraction removes not only essential oil from the material, but also
other non-volatile constituents and these are retained in the product in the heating process and
tend to fix or stabilize the most volatile essential oil (Eiserle & Rogers, 1972).
In order to obtain an ideal formula, the cosmetic must be stable, and for this to happen
the evaluation of the stability of the products must be one of the most important factors in the
execution of the processes involved (Zanin, Miguel, Chimell, & Dalmaz, 2001). Therefore,
the objective of this study was to obtain cinnamon oleoresin, as well as to evaluate the
antimicrobial properties of the oleoresin and the essential oil and employ them as additives in
the preparation of organic lip balms enriched with vitamin E. In addition, the accelerated and
extended stability analysis, as well as the microbiological safety assessment of the products.
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
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Therefore, the present study aimed to use essential oil and cinnamon oleoresin in the
production of lip balms, evaluating whether these additives differ in the microbiological
stability and safety of the cosmetic product as suggested by Eiserle & Rogers (1972) for food.
2.Metodology
Obtaining the oleoresins: The cinnamon oleoresin obtention were carried out using
30 grams of the material and 350 mL of absolute anhydrous alcohol (99.3º) in a Soxhlet
apparatus for two hours at 80C. After extraction, the solvent was concentrated using a rotary
evaporator.
Antimicrobial activity of the oleoresins and essential oils: The cinnamon essential
oil was purchased from Ferquima Industria e Comércio Ltda. Different microorganisms were
used, as Gram-negative bacteria E. coli and as Gram-positive bacteria S. aureus and B. cereus.
The bacterial cultures were diluted to a final concentration of 106 cells/mL and 500μL of these
solutions were spread on Petri dishes, separately, containing Muller Hinton Agar. Over the
broth were deposited the oleoresin and essential oil under study. The Petri dishes were
incubated at 37°C for 24 h and after that, the size of inhibition zones was measured in
millimetres. These analyses were performed in triplicate.
Gas Chromatography coupled to Mass Spectroscopy (GC-MS) of Cinnamon
essential oil and oleoresin: according to Eiserle & Rogers (1972) oleoresin have in their
constitution essential oils and other compounds, of a non-volatile nature, that act as fixatives
and tend to depress the volatilization of essential oils during thermal processing. Thus, an
evaluation by Gas Chromatography coupled with Mass Spectrometry of the essential oil and
oleoresin used in this work was carried out to investigate their chemical composition. For this,
1 g of the samples was dissolved in 10 ml of HPLC grade acetone. The solution was vortexed
and kept in an ultrasonic bath for 10 minutes. Then the samples were filtered through a 0.45
μm RC filter before analysis. The analyses were performed in a High-Resolution Gas
Chromatograph coupled to a Mass Spectrometer Detector - Shimadzu, model GC / MS-
QP2010. GCMS, using an Agilent DB-5MS column (30 m x 0.25 mm - 0.25 μm). The
equipment conditions were: Injector temperature at 220°C, Splitless Injection Mode with 2-
minute sampling time, Linear speed flow control mode, 15.7 psi pressure, total flow of 19.4
mL. min-1, Flow in the column of 1.49 ml.min-1, Linear speed of 45.0 cm.sec-1, Purge flow of
3.0 ml.min-1, Split ratio of 10, Column temperature in Thermogradient mode (80C for 2
minutes, 140C and 280C). The Parameters of the Mass Spectrometry Detector were: Ion
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
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source temperature at 200°C, Interface temperature at 280°C, Solvent cut-off time of 3
minutes, Detector voltage in relation to the result of the Tuning, Initial detection in 3.5
minutes, Final detection time in 17.0 minutes, SCAN acquisition mode, 0.25 second
acquisition time, SCAN mass/load ratio (m/z) from 40 to 600 and volume of injection of 1
μL.
Preparation of lip balms: 45 g of coconut oil, 24 g of Candelilla wax and 45 g of
cupuaçu butter were weighed, taken to the water bath at 100°C to form a homogeneous
mixture which was given the named base. As the objective was the analysis of the influence
of cinnamon oleoresin/essential oil and vitamin E to the base, 6 different types of lip balms
were prepared varying the compositions. In all variations 18 g of base were used. For the
balms containing vitamin E, four capsules of the vitamin were added in each sample. To the
balms containing the oleoresin or essential oil, 5 drops were added in each variation. All the
samples prepared were stored in an acrylic container, properly labelled according to the
following nomenclature and composition:
• Control 1: base;
• Control 2: base + vitamin E;
• Control 3a: base + Cinnamon oleoresin;
• Control 3b: base + Cinnamon essential oil;
• Sample 1: base + vitamin E + Cinnamon oleoresin;
• Sample 2: base + vitamin E + Cinnamon essential oil.
Balm Stability Tests: Two stability tests were carried out on the prepared balms, the
Accelerated Stability Test (AST) and the Extended Stability Test (EST). The AST lasts 15
days and the EST lasts 90 days. For this, the test formulations were subjected to stress
conditions in order to accelerate the appearance of possible signs of instability according
(ANVISA, 2012). The samples in this study were kept in the following environments and
temperatures:
• Oven: T = 40C;
• Refrigerator: T = 5C;
• Freezer: T = –5C;
• Cycles: 24 h at 45C and 24 h, at –5C;
• At 25C under UV light;
• At 25C protected from UV light.
The entire stability assessment was carried out in triplicate for the environment and
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
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temperature under study. The analyses were carried out every 3 days over a period of fifteen
days (AST) and every 15 days for 90 days (EST), investigating the organoleptic
characteristics, which are those perceptible by the human senses. The appearance, colour and
odour of the prepared balms were analysed, which can be classified as normal or unchanged
(N); slightly modified (SM); modified (M) and intensely modified (IM).
Evaluation of Microbiological Safety of the products: for the evaluation of the
microbiological safety of the balms, the research of Escherichia coli was carried out, as well
as, of Staphylococcus and Pseudomonas, according to ANVISA (2012). For E. coli research,
1 g of sample was aseptically transferred to 9 mL of saline. Then, 1 ml of the solution was
transferred to the Petri dish containing Eosin Methyl Blue agar (EMB) and the dish was
incubated at 36 ± 1ºC for 24 hours. After this period, it was observed whether there was a
growth of colonies, as well as their characteristics (if present, they are black metallic
colonies). When there is growth of suspicious colonies, the biochemical series for the
identification of E. coli must be followed. For the research of Staphylococcus and
Pseudomonas, 1 g of sample was aseptically transferred to 9 mL of soy-casein broth and this
broth was incubated at 36 ± 1 ºC for 24 to 48 hours. Then, it was sown in Petri dishes
containing Vogel Johnson agar for Staphylococcus research, and in cetrimide agar plates for
Pseudomonas research. The plates were incubated at 36 ± 1 ºC for 24 hours. After this period,
it was observed whether there was growth and the characteristics of the colonies. For
Staphylococcus, the presence of yellowish colonies or the appearance of specific
characteristics must be confirmed with the catalase and coagulase tests. For Pseudomonas, on
the other hand, the presence of blue-green colonies must be confirmed by the microorganism
by the biochemical series.
3.Results and Discussion
Antimicrobial activity of the cinnamon oleoresin and essential oil: Cinnamon
oleoresin and essential oil inhibited entirely the growth of the microorganism tested. These
results are important to verify if it is necessary a high antimicrobial activity to be used in a
cosmetic product to inhibit the growth of pathogenic microorganisms.
GC-MS of Cinnamon essential oil and oleoresin: According to the GC-MS analysis,
the cinnamon essential oil and oleoresin studied are completely different both in the amount
of compounds present and in the nature of the composition itself. GC-MS of cinnamon
essential oil showed 11 peaks between 3.2 and 14.3 minutes (Figure 1), while cinnamon
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
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oleoresin showed 5 peaks between 1.2 and 13.4 minutes (Figure 2). The compounds present
in essential oil and cinnamon oleoresin and their chemical structures are listed in each
chromatogram.
Figure 1: Chromatogram and compounds present in cinnamon essential oil studied.
OH
H
O
O
O
OH
O
O
O
OH
O
O
O
Source: Authors.
Da Silva et al. (2012) studied the biological activity of pinenes and demonstrated that
both enantiomers, α- and β-pinene have antimicrobial activity. However, only (+) enantiomers
show such activity. According to Dorman & Deans (2000), alcohols have bactericidal activity
against vegetative cells. Still according to the authors, aldehydes have high antimicrobial
activity, possibly due to the conjugation of the aldehyde group to a C=C double bond in a
highly electronegative arrangement. The authors also suggest that an increase in
electronegativity improves antimicrobial activity, since electronegative compounds can
interfere with biological processes by electron transfer and react with nitrogenous
components, such as proteins and nucleic acids, inhibiting the growth of microorganisms.
According to Hyldgaard et al. (2012), although the mode of action of cinnamaldehyde is
inconclusive, aldehydes covalently bind to the amino groups of DNA and proteins, reducing
their normal function. Cinnamaldehyde still interacts with the cell membrane.
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
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Figure 2: Chromatogram and compounds present in cinnamon oleoresin studied.
OH
O
H
O
OO
Source: Authors.
For Goodarzi, Hadjiakhoondi, Yassa, Khanavi & Tofighi (2016) heterocyclic
compounds containing the methylenedioxy functional group, such as 5-(2-propenyl) -1,3-
benzodioxol also known as safrole, have a wide range of biological activities, such as
antitumor, antibacterial, antifungal, antiparasitic, antimalarial, antioxidant, pesticide and
herbicide. For Shah & Shelar (2018), both 2-methoxy-3-(2-propenyl)-phenol and 2-methoxy-
4-(2-propenyl)-phenol also have antimicrobial activity. De Campos (2017) reports the
biological activity of benzyl benzoate, present in extracts of green propolis. In the review on
coumarin antimicrobial activity written by Smyth et al. (2009), the authors showed that these
compounds are selective for Gram-positive microorganisms.
Taking into account cinnamon oleoresin, cinnamaldehyde, alcohols and coumarin are
reported as antimicrobial compounds. But since cinnamaldehyde is the major compound, it
probably is the responsible for the antimicrobial activity. Considering cinnamon essential oil,
α-pinene, o-cymene, β-linalool, cinnamaldehyde, 5-(2-propenyl)-1,3-benzodioxol, 2-
methoxy-3-(2-propenyl)-phenol, 2-methoxy-4-(2-propenyl)-phenol and benzyl benzoate, have
antimicrobial activity reported in the literature. Although, as 2-methoxy-3-(2-propenyl)-
phenol it is the major compound in the sample, it is probably the responsible for the
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
10
antimicrobial activity. Thus, this study goes against the one proposed by Eiserle & Rogers
(1972), as it is possible to verify by the results obtained that the compounds present in
cinnamon essential oil are not found in cinnamon oleoresin. Only cinnamaldehyde coexists in
both essential oil and oleoresin. It is noteworthy that, there was no analysis of the non-volatile
compounds present in the studied oleoresin to prove the existence of such compounds and to
know their composition.
Stability Tests with the balms under study: When performing the AST, variations in
the organoleptic properties were observed in the evaluation of aspects, colour and odour in the
studied samples, performing each test in triplicate for the two comparisons, with oleoresin and
essential oil. The results of the tests are shown in Table 1. As can be observed from Table 1,
both samples with essential oil and with oleoresin at 25C exposed to UV light were more
unstable. The characteristics observed in the product with essential oil were slightly altered in
the sixth and fifteenth day of evaluation, with the presence of water droplets in the three
samples. For the oleoresin samples, instability also started to appear on the sixth day of
evaluation, with the presence of water droplets in the samples (presence of condensation),
showing a slight increase in this aspect over the following days. Another slightly moderate
instability observed in the aspect analysis was the presence only on the fifteenth, day of water
droplets in the samples with oleoresin at 25C without UV light. The explanation for this
effect in the samples, is because the air present between the lids and the surface of the
products was saturated with water vapor that ended up condensing.
It should be noted that there were no cracks in the samples analysed and it is believed
that this occurred due to the presence of vitamin E in the formulations. This is because,
according to Rowe, Sheskey and Quinn (2009) and Barel, Paye and Maibach (2009), vitamin
E acts attracting the water molecules, acting as a humectant. Therefore, as there was vitamin
E in these formulations, the water molecules present in the products did not dissipate to the
external environment, which in turn did not cause the appearance of cracks.
Research, Society and Development, v. 9, n. 9, e539997544, 2020
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Table 1. Accelerated Stability Test data.
Control 1
Control 2
Control 3A
Control 3B
Sample 1
Sample 2
Storage time (days)
0
3
6
9
1
2
15
0
3
6
9
12
15
0
3
6
9
12
15
0
3
6
9
12
15
0
3
6
9
1
2
1
5
0
3
6
9
1
2
1
5
Freezer
-5C
Aspect
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
S
M
S
M
S
M
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Odour
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Refrigerator
5C
Aspect
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
S
M
S
M
S
M
L
M
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Odour
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Cycle
Aspect
N
N
N
S
M
S
M
M
N
N
N
N
N
N
N
N
N
M
S
M
M
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
N
N
S
M
S
M
S
M
N
N
N
S
M
S
M
S
M
N
N
N
S
M
S
M
S
M
N
N
N
S
M
S
M
S
M
N
N
N
S
M
S
M
S
M
N
N
N
S
M
S
M
S
M
Odour
N
N
S
M
N
N
N
N
N
S
M
N
N
N
N
N
M
N
N
M
N
N
S
M
N
N
S
M
N
N
S
M
N
N
N
N
N
S
M
N
N
N
25C without
UV light
Aspect
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
N
N
N
N
S
M
N
N
N
N
N
S
M
N
N
N
N
N
S
M
N
N
N
S
M
N
S
M
N
N
N
N
N
S
M
N
N
N
N
N
S
M
Odour
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
M
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
25C with UV
light
Aspect
N
N
N
N
S
M
N
N
N
N
N
S
M
N
N
N
N
N
N
N
N
N
S
M
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
N
M
M
S
M
M
N
N
M
M
S
M
M
N
N
M
M
S
M
M
N
N
M
M
S
M
M
N
N
M
M
S
M
M
N
N
M
M
S
M
M
Odour
N
N
S
M
N
N
N
N
N
S
M
N
N
N
N
N
M
N
N
S
M
N
N
S
M
N
N
N
N
N
S
M
N
N
N
N
N
S
M
N
N
N
Oven
Aspect
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
N
N
S
M
S
M
S
M
N
N
N
S
M
S
M
S
M
N
N
N
S
M
S
M
S
M
N
N
N
S
M
S
M
S
M
N
N
N
S
M
S
M
S
M
N
N
N
S
M
S
M
S
M
Odour
N
N
S
M
S
M
N
N
N
N
S
M
S
M
N
N
N
N
S
M
S
M
N
M
N
N
S
M
S
M
N
N
N
N
S
M
S
M
N
S
M
N
N
S
M
S
M
N
N
Legend: N - Normal, SM - Slightly Modified, M - Modified, IM - Intensely Modified. Source: Authors.
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
12
For the colour analysis, no variations were observed for samples kept at low
temperatures, such as in the freezer (-5C) and refrigerator (5C) for the two comparisons. In
the remaining conditions, samples and controls with oleoresin showed instability at 25C with
UV light, demonstrating a slightly moderate change to a lighter colour on the sixth and ninth
test days and remaining stable from the fifteenth day. Another observation is a small
modification on the fifteenth day for the cycle control, which showed a darker colour. For the
samples with essential oil, changes were observed in the controls stored under UV light, with
instability on the sixth and ninth day and in the controls at 25C without UV light, only on the
ninth day, with a slight lightening of the sample as an alteration. For the control 1 maintained
in the cycle condition, a darkening was observed on the sixth day, which remained constant in
the next days of analysis for the three samples.
Some ingredients, especially colouring solutions and pigments can change their colour
due to oxidation reactions that occur when they are exposed for a long time to oxygen. In
addition, products that contain oily-based materials in their formulation and are exposed to air
or extreme temperatures can degrade by oxidation (Galembeck & Csordas, 2009). In such a
way, it can be concluded that the samples and controls that were present in the conditions of
cycles and at 25C under UV light underwent oxidation of the compounds present in the
candelilla wax. Another factor also responsible for the oxidation reaction is the action of UV
light (Galembeck & Csordas, 2009). From this it is possible to affirm the samples and
controls that were exposed to solar radiation suffered oxidation, therefore, causing their
colour variation. It shows the exposure condition was a factor that caused a slow rate of
degradation of the compounds present in the product formulations, mainly those present in
candelilla wax. This slow rate of degradation is due to the presence of vitamin E in the
formulations, an antioxidant.
The last organoleptic characteristic analysed was the odour of the studied samples.
Analysing the two sets, instabilities were observed when the samples were exposed to higher
temperatures. Evaluating the first set, the changes appeared from the twelfth day of storage at
25C with UV light and on the fifteenth day for samples in the oven (40C). For the second
set, the instabilities arose from the ninth day for tests at 25C with UV light and on the
fifteenth day for samples stored in an oven (40C). All the mentioned samples had a slightly
moderate instability change, with attenuation of the characteristic odour of cinnamon and
started to have a slight odour of candelilla wax. When an oil-based product is exposed to air
or extreme temperatures, it can degrade by oxidation, generating the unpleasant rancid odour.
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
13
Taking this aspect into account, vitamin E was added to the formulation as an antioxidant
(Galembeck & Csordas, 2009). Thus, no rancid odour was observed in any of the controls or
samples analysed and as only the attenuation of the characteristic odour of cinnamon was
observed to accentuate the characteristic odour of candelilla wax, it is possible to conclude
that vitamin E acted in reducing the effects caused by the oxidation reaction, one of which is
the rancid odour.
However, as can be seen from the results obtained in the stability tests performed,
there was no difference in the stability of the samples in the two sets analysed. Therefore, it is
possible to affirm that, although oleoresin does not contain the same compounds present in
the essential oil of cinnamon used in this work, the compounds present in oleoresin were
sufficient to maintain the same stability observed for the samples with essential oil. Thus, it is
possible to affirm that although they do not have the same chemical composition, both
cinnamon oleoresin and essential oil are additives that can be used in the manufacture of
cosmetic products, with antimicrobial and preservative action.
To complement the AST, the EST was performed following the same procedures as
the AST test, removing the samples kept in a condition of temperature cycles. The results
obtained from such an evaluation are shown in Table 2. For the analysis related to the aspect,
a small modification was noted for the samples with essential oil on the ninetieth day of the
test for low temperatures such as freezer (-5C) and refrigerator (5C) and for the test in the
oven (40C). All of the aforementioned had a slightly drier appearance than the reference,
stressing that there were no cracks, probably due to the presence of vitamin E in the samples,
proving that the release of water molecules present in the products happened very slowly and
gradually. For the sample at 25C with UV light, instability was observed from the seventy-
fifth day with the disappearance of water droplets but forming a slightly moderate to
oleaginous aspect. Finally, the samples with oleoresin showed the same mildly dry
appearance on the ninetieth day for freezer storage, but without cracks, and for the samples at
25C without UV light the formation of small water droplets was noticed on the last
evaluation day. For the sample exposed to UV light, the presence of water droplets and a
slightly more oleaginous aspect was also noticed from the seventy-fifth day, which proves
that the air present between the caps and the product surfaces was saturated with steam water
and its condensation happened gradually.
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
14
Table 2. Extended Stability Test data.
Control 1
Control 2
Control 3A
Control 3B
Sample 1
Sample 2
Storage time (days)
0
1
5
3
0
45
60
75
90
0
1
5
30
4
5
60
75
90
0
15
30
45
6
0
7
5
9
0
0
1
5
3
0
4
5
6
0
7
5
9
0
0
1
5
3
0
4
5
6
0
7
5
9
0
0
1
5
3
0
4
5
6
0
7
5
9
0
Freezer
-5C
Aspect
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
S
M
S
M
S
M
S
M
S
M
S
M
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Odour
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Refrigerator
5C
Aspect
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
S
M
S
M
S
M
S
M
S
M
S
M
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Odour
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
25C without
UV light
Aspect
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
S
M
S
M
S
M
S
M
S
M
S
M
N
N
S
M
S
M
S
M
S
M
S
M
N
S
M
S
M
S
M
S
M
S
M
S
M
N
S
M
S
M
S
M
S
M
S
M
S
M
N
S
M
S
M
N
S
M
N
S
M
N
S
M
S
M
S
M
S
M
N
S
M
Odour
N
N
N
N
S
M
N
S
M
N
N
N
N
S
M
N
S
M
N
S
M
N
N
S
M
N
S
M
N
N
N
N
S
M
N
S
M
N
N
N
N
S
M
N
S
M
N
N
N
N
S
M
N
S
M
25C with UV
light
Aspect
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
M
S
M
S
M
S
M
S
M
S
M
N
N
S
M
S
M
S
M
S
M
S
M
N
N
S
M
S
M
S
M
S
M
S
M
N
N
S
M
S
M
S
M
S
M
S
M
N
N
S
M
S
M
S
M
S
M
S
M
N
N
S
M
S
M
S
M
S
M
S
M
Odour
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
S
M
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Oven
Aspect
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
Colour
N
S
M
S
M
S
M
S
M
S
M
S
M
N
N
S
M
S
M
S
M
S
M
S
M
N
N
S
M
S
M
S
M
S
M
S
M
N
N
S
M
S
M
S
M
S
M
S
M
N
N
S
M
S
M
S
M
N
S
M
N
N
S
M
S
M
S
M
S
M
S
M
Odour
N
N
N
S
M
S
M
N
S
M
N
N
N
N
S
M
N
S
M
N
S
M
S
M
N
S
M
N
S
M
N
N
N
N
N
N
S
M
N
S
M
N
N
N
N
S
M
N
N
N
N
N
N
S
M
Legend: N - Normal, SM - Slightly Modified, M - Modified, IM - Intensely Modified. Source: Authors.
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
15
For the colour analysis of the samples, it was observed a gradual variation in the
analysed products. Control A showed a slight colour change on the ninetieth day of storage in
an oven (40C) and at 25C without UV light, showing a slightly darker colour than the
reference. For the sample at 25C with UV light, a progressive loss of colour was observed
from the thirtieth day, showing a slightly white colour at the end of the test. The control 2 that
found a more significant colour change were for samples exposed to sunlight gradually losing
their colour at high temperature. Therefore, observing the results, the oxidation occurred
slowly and gradually and can be justified due to the addition of vitamin E in the formulations.
Finally, the odour of the samples showed mostly mild changes in the last days of evaluation
of the controls, thus losing the natural aroma of their essences (cinnamon). However, in the
last organoleptic analysis, the odours did not coincide mostly with the odour of the samples at
the beginning of the test, proving that oxidation, a probable cause of the colour change, is not
the only responsible for the odour change, but also the volatility of the components present in
the formulation of the products.
Evaluation of Microbiological Safety of the products: According to the
microbiological safety assessment of the products studied, there was no E. coli, just as there
was no Staphylococcus and Pseudomonas growth. In this way, it is possible to conclude that
both essential oil and cinnamon oleoresin are equally effective antimicrobial additives, as
none of the products showed growth of pathogenic microorganisms in the researches carried
out compared to control 1. The different chemical composition of the essential oil and
cinnamon oleoresin studied did not compromise the antimicrobial action of these additives.
Therefore, it can be concluded that the fact that oleoresin does not contain all the compounds
present in the essential oil did not compromise the antimicrobial action of this additive. In
other words, is possible to say that the synergistic action of the various compounds present in
the essential oil does not increase its antimicrobial action and that, probably, cinnamaldehyde
is the compound responsible for the antimicrobial activity of both the essential oil and
oleoresin, or if this is in greater quantity, this may also be the possible explanation for the
activity in both additives. According to Orth (1984) all cosmetic products are subject to
contamination with microorganisms. However, the growth of fungi, bacteria and spores in
cosmetic products depends on a series of physical and chemical factors, such as the
availability of water, the composition of the product that can serve as nutrients for
microorganisms, storage temperature, among others. For Carvalho & Pachione (1989), the
lack of hygiene in manufacturing and the low stability of the constituents of the formulation
are the main factors that contribute to the contamination of non-sterile products. However,
Research, Society and Development, v. 9, n. 9, e539997544, 2020
(CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v9i9.7544
16
according to McCarthy (1984), microbial contamination can cause visible changes, such as
changes in colour, odour, viscosity, in addition to being able to cause toxic reactions to the
user.
4. Conclusions
Due to the growing consumption of cosmetic products, the industries of the sector, in
addition to increasing their production lines and also meeting the criteria imposed by the
quality agencies, need to improve their products, so new research for the development and
formulation is being performed. The products formulated in this work met the criteria related
to stability tests, both accelerated and extended, even with variations found in the products,
these did not interfere in the synergistic relationships of its components, leaving them stable.
The microbiological evaluation of the formulated products showed that the formulated
products are 100% microbiologically safe.
In future studies other essential oils and oleoresins will be used to test the same properties.
Acknowledgements
The authors thank FAPEMIG, CNPQ and CAPES.
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Porcentagem de contribuição de cada autor no manuscrito
Leonam José de Andrade Lima – 15%
Luana Cominato – 15%
Heloísa Moraes Oliveira – 15%
Wanderson de Oliveira dos Santos – 15%
Geoffroy Roger Pointer Malpass – 10%
Mônica Hitomi Okura – 10%
Ana Claudia Granato – 20%
