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Caffeic acid: a review of its potential use in
medications and cosmetics
C. Magnani,*V. L. B. Isaac, M. A. Correa and H. R. N. Salgado*
Besides powerful antioxidant activity, increasing collagen production and prevention of premature aging,
caffeic acid has demonstrated antimicrobial activity and may be promising in the treatment of dermal
diseases. The relevance of this study is based on the use of caffeic acid that has become increasingly
common in humans. Thus, studies that demonstrate and clarify the functions of this substance are very
important.
Introduction
Characteristics of caffeic acid
Phenolic compounds occur universally in the plant kingdom
and are part of a large and complex group of organic
substances. Higher plants synthesize and accumulate a wide
variety of phenolic compounds, which confer protection against
the attacks of free radicals, which are by-products from the
process of photosynthesis, and against tissue injuries.
1
The phenolic compounds can be classied into two groups: the
group of simple phenolic compounds and the group of poly-
phenolic compounds which can be observed through the frame
shown in Fig. 1.
2,3
Cinnamic acid derivatives, also called phenylpropanoids, are
nine carbon structures (Fig. 2). Caffeic acid (3,4-dihydroxycin-
namic) is representative of this group.
Modern consumers increasingly demand products and foods
with high quality. Cosmetics may not have as many health
implications as pharmaceutical products or food, but they are
chemical products that people use daily and apply on skin, hair,
nails and even teeth or mouth.
4
Allied to this, legislation is
increasingly strict about quality and safety, which has chal-
lenged the industries.
5
For this reason, natural antimicrobial
compounds, such as polyphenols,
6
are being widely used.
Caffeic acid (3,4-dihydroxycinnamic) is one of the hydroxy-
cinnamate and phenylpropanoid metabolites more widely
distributed in plant tissues. This polyphenol is present in many
food sources, including coffee drinks, blueberries, apples and
cider.
7
Besides food, caffeic acid is present in several medica-
tions of popular use, mainly based on propolis.
8
Besides acting
as a carcinogenic inhibitor,
9,10
it is also known to possess anti-
oxidant and antibacterial activity in vitro, and can contribute to
the prevention of atherosclerosis and other cardiovascular
diseases.
11–13
Antioxidant activity
Antioxidants are compounds that inhibit or reduce the effects
triggered by free radicals and oxidizing compounds.
14
Phenolic antioxidants act as free radical scavengers and
sometimes as metal chelators. They act both in the initiation
step and in the propagation of the oxidative process. The
products are relatively stable due to the resonance of the
aromatic ring shown by these substances. The phenolic acids
Fig. 1 Chemical classification of phenolic compounds.
Fig. 2 Chemical structure of the main cinnamic acids. Cinnamic acid
R1 ¼R2 ¼R3 ¼R4 ¼H, o-coumaric acid: R2 ¼OH, p-coumaric acid:
R3 ¼OH; caffeic acid: R2 ¼R3 ¼OH; ferulic acid: R2 ¼OCH
3
and
R3 ¼OH.
UNESP –Universidade Estadual Paulista, Faculdade de Ciˆ
encias Farmacˆ
euticas,
Departamento de F´
armacos e Medicamentos, Araraquara, SP, Brazil. E-mail: carol.
magnani@hotmail.com; salgadoh@fcfar.unesp.br
Cite this: Anal. Methods,2014,6,3203
Received 15th October 2013
Accepted 22nd February 2014
DOI: 10.1039/c3ay41807c
www.rsc.org/methods
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are characterized by the presence of a benzene ring, a carboxylic
acid group and one or more hydroxyl or methoxy groups, which
confers antioxidant properties.
15
The phenylpropanoids act as antioxidants by eliminating
oxygen free radicals
16,17
and chelating pro-oxidant metal ions,
especially iron.
18,19
The hydroxyl groups of these molecules
confer antioxidant activity, but they are not the only factors in
determining the potency of their activities. There is a single
hydroxyl group para-substituted in an aromatic ring that is
linked to a side chain conjugate in the case of ferulic acid. This
substitution allows the phenoxy radical free electrons to
become delocalized over the entire molecule, and therefore
stable.
20
The ortho substitution to the methoxy group, an elec-
tron donor, is also a contributing factor to the stability of the
phenoxy radical and thus increases the antioxidant effi-
ciency.
21,22
The presence of a second hydroxyl group in the ortho
position, besides the para position, is known to increase the
antioxidant activity due to an additional resonance stabilization
and formation of o-quinone (Fig. 3).
20,21,23
This characteristic
can be used to explain the fact that the efficiency of antioxidants
such as caffeic acid is greater than that of ferulic acid.
Caffeic acid has been shown to have protective effects on a-
tocopherol in low-density lipoprotein (LDL).
24
Furthermore, its
combination with other products, such as chlorogenic and
caaric acids, showed more potent antioxidant activity in a
variety of different systems.
17,25
Natural phenolic antioxidants including caffeic and ferulic
acids gained remarkable attention as promising photo-
protective agents
26,27
which have also been used in skin care
products due to their antioxidant activity. However the litera-
ture shows little evidence about the usefulness of hydroxycin-
namic acids in protecting the skin from photo-oxidative
damage.
The normal cellular metabolism produces free radicals,
including reactive species of oxygen and nitrogen, which are
derived from both normal metabolic processes and essential
metabolic processes in the body (endogenous), and they can be
derived from exposure to environmental factors (exogenous)
such as pollution, radiation, pesticides, and tobacco, among
others. Free radicals can cause benecial or deleterious effects
to health.
28–30
The importance of reactive oxygen species (ROS) and free
radicals has attracted increasing attention over the last decade.
ROS include free radicals such as superoxide anion radicals
(O
2
c
), hydroxyl radicals (OHc) and reactive oxygen species such
as H
2
O
2
and singlet oxygen (1O
2
). These molecules exacerbate
factors of cell damage and aging.
31,32
ROS are continuously produced during normal physiological
events and they can easily initiate the peroxidation of
membrane lipids, leading to the accumulation of lipid perox-
ides.
33,34
However antioxidant cells have developed mechanisms
to protect themselves from free radical toxicity. The agents
considered to be antioxidants include catalytic enzymes that
remove radicals such as superoxide dismutase (SOD); proteins
which minimize the availability of pro-oxidants, such as iron
ions and copper ions, for example, transferrins, ferritins, met-
allothionein and haptoglobines; and low molecular weight
molecules that have the ability to capture reactive oxygen
species through autoxidation such as, for example, those with
glutathione and thiol groups (SH) or vitamins such as a-
tocopherol, ascorbic acid and b-carotene.
35,36
When there is an imbalance between pro and antioxidant
systems, with a predominance of oxidants, the oxidative stress
occurs.
37,38
This stress can be associated with damage to lipids,
proteins and genes and is implicated in a great variety of human
diseases as well as aging.
39
The free radicals also affect the skin tissue, which originate
mainly from exogenous actions, such as ultraviolet radiation.
Depending on the dose, exposure time, wavelength and area
exposed, the ultraviolet radiation can cause skin burns,
premature skin aging and even skin cell DNA damage and skin
cancer.
40,41
Thus, the natural aging of the skin associated with the action
of free radicals causes a reduction in skin hydration, pigmen-
tation, ne wrinkles, signals from sagging and increased
possibility of the occurrence of neoplasm diseases. Another
change is the reduction of dermal collagen that makes the skin
thinner. These changes are even more evident aer menopause,
where there is a rapid decrease in the levels of collagen in skin
and bone, suggesting that estrogen inuences the collagen
synthesis in the skin as much as bone mass.
42
In order to maintain the skin healthier and younger-looking,
thereby mitigating the effects of aging, more strategies have
been suggested.
30
The use of natural or synthetic antioxidants in
foods, cosmetics, beverages and also medicine is one of the
defence mechanisms against free radicals.
43–45
Photoaging of the skin is one of the most common derma-
tological concerns and can become a major health concern
because it is correlated with an increased risk of skin cancer.
42
UVA radiation can cause clinical, biochemical and histological
changes in the skin through changes in cells and in extracel-
lular matrix proteins, including collagen, responsible for the
structural integrity of the skin.
46
UVA radiation (315–400 nm) has been shown to elevate
matrix metalloproteinase-1 extracellular (collagenase MMP-1)
and most proteolytic enzymes produced by the skin cells,
including keratinocytes and broblasts.
47,48
This activation is
related to oxidative stress, which occurs when there is excessive
production of reactive oxygen species and/or depletion of anti-
oxidant defence in cells.
49,50
Natural and synthetic antioxidants act as scavengers of free
radicals and have been shown to promote skin photo protection
Fig. 3 Chemical structure of caffeic acid and formation of ortho-
quinone.
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by inhibiting the induction of MMP-1 activity mediated by
UVA radiation and expression of keratinocytes and skin
broblasts.
48,51,52
Ultraviolet A (UVA) plays a fundamental role in the patho-
genesis of premature skin aging through cytotoxicity to kera-
tinocytes and degradation of collagen, a major component of
the extracellular matrix that provides structural support.
Protection against UVA damage mediated through the anti-
oxidant defence systems has been proposed as a possible
mechanism by which plant compounds decrease the process
of premature aging.
53
Furthermore, under stimulation by UVA and UVB sunlight,
keratinocytes from human skin can secrete nitric oxide (a
radical involved in oxidation reactions). Nitric oxide (NO)
appears to have great interest in the formation of erythema and
skin inammation.
54,55
Both caffeic and ferulic acids act as
sequestering agents for radical NO.
56
In light of these observa-
tions, the topical administration of antioxidants found
considerable interest, since it represents an effective strategy to
protect the skin against oxidative damage mediated by UV.
55,57
The extension of erythema caused by UVB radiation may be
monitored through reectance spectrophotometry, which is
considered one of the most suitable models for the in vivo study
of skin damage aer acute exposure to UV.
58,59
Caffeic and
ferulic acids, dissolved in saturated aqueous solutions at pH
7.2, proved to provide signicant photo-protection to the skin
against UV-induced erythema.
56
Anti-aging products are being increasingly used in recent
years and those with active antioxidants in their composition
are highlighted.
60
There are various methods for determining the in vitro
antioxidant activity of substances, such as: ferric thiocyanate
method, method of inhibition of ABTSc
+
and DPPHc, the
superoxide anion radical and metal chelating activity. In all
these methods, caffeic acid proved to be an effective antioxidant
when compared with conventional antioxidants such as BHA,
BHT, a-tocopherol, and Trolox, a water soluble analogue of
tocopherol.
61
Through the permeation study of caffeic acid, chlorogenic
acid and oraposide, it was found that the caffeic and chloro-
genic acids are able to permeate all the skin layers of pig ear,
which have a systemic activity, whereas oraposide remained in
the upper layer of the skin surface.
62
Permeation studies on
human skin also conrm the ability of caffeic acid and ferulic
acid to permeate into the skin, a characteristic independent of
the pH of the receptor solution being 3.0 or 7.2.
63
As the solar radiation penetrates deeply into the skin, it is
necessary to ensure that topically applied substances are able to
penetrate through the stratum corneum, the main barrier against
permeation of foreign substances through the skin and reach
the deepest layers to promote satisfactory photo-protection.
64
The absorption of a compound in the skin is determined by
its physicochemical characteristics, and on the permeation
process, the lipophilicity is one of the most important
features.
65
Thus, the high lipophilicity of caffeic and ferulic
acids may explain the fact that they permeate through the
stratum corneum.
66
The ferulic and caffeic acids also demonstrate protective
effects against phosphatidylcholine peroxidation induced by UV
radiation, which is important since the phosphatidylcholine is a
major constituent of the lipid bilayers of cell membranes.
56
Antimicrobial activity
Every day more consumers seek more effective cosmetics, which
approximate the denition of cosmeceuticals, or actually having
some biological effects, particularly in the prevention of
premature aging and diseases such as cancers caused by the
action of free radicals in the genes of cells. Therefore, similar to
drugs, these products should also provide security to the
consumer. Allied to this, legislation is becoming increasingly
stringent, which has challenged the industries.
4,5
Besides,
cosmetic companies are required to control the optimal pres-
ervation of their commercial products, since microbial
contamination in cosmetics represents an important risk for
consumer health.
67
For this reason, natural antimicrobial
compounds are widely used.
6
Besides its remarkable antioxidant activity, in vitro studies
have demonstrated antimicrobial properties of propolis against
various oral pathogens. Several components of propolis have been
analysed in different countries, and caffeic acid, phenethyl ester
of caffeic acid and avonoids are the main ingredients respon-
sible for the antibiotic power of this resin.
68–70
Thus, a thorough
search of the antimicrobial activity of caffeic acid is promising
targeting the treatment of dermal diseases, such as acne.
Some phenylpropanoids, including caffeic acid, p-coumaric
acid and ferulic acid, are able to inhibit the growth of bacteria,
including E. coli,Staphylococcus aureus,Bacillus cereus,Listeria
monocytogenes and some yeasts.
71
The antimicrobial activity of these substances depends not
only on their structure but also on the environmental condi-
tions. Caffeic acid has a reduced ability to inhibit Listeria
compared to other hydroxycinnamic acid derivatives due to the
presence of a high degree of hydroxylation.
6
Moreover, envi-
ronmental factors, including the pH and concentration of
sodium chloride, are important because they produce certain
physiological changes in the micro-organisms that make them
more sensitive to phenylpropanoids.
72
Antioxidant and antimicrobial activities of caffeic acid are
pronounced in cosmetic emulsions with acidic pH (3–5). Caffeic
acid showed, at this pH, antimicrobial effects against various
micro-organisms studied (E. coli,Pseudomonas aeruginosa,
Bacillus cereus,Kokuria rhizophila,Staphylococcus aureus,Listeria
monocytogenes,Candida albicans).
73
The mechanism of the antimicrobial activity of substances is
established considering one of the following three: (1) reaction
with the cell membrane causing increased permeability and
loss of cellular constituents, (2) inactivation of enzymatic
systems or essential enzymes, including those involved in the
production process energy and synthesis of structural compo-
nents, or (3) destruction or inactivation of functional genetic
material.
74,75
The cytoplasm membrane of bacteria and the mitochondrial
membrane of yeasts are permeable barriers to passage of small
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ions such as H
+
,K
+
,Na
+
and Ca
2+
, in addition to being
responsible for the input and output of different compounds.
This cell permeability is important for various cell functions
such as the maintenance of energy in the transduction process,
solute transport, metabolism regulation and pressure
control.
76,77
There is a consensus that aromatic and phenolic
compounds, such as caffeic acid, affect the cytoplasm
membrane, alter its structure and function, change the active
transport and coagulate cell content.
78,79
Cytotoxicity
Caffeic acid and its derivatives, such as caffeic acid phenyl ester,
have action against colon and oral cancer, and they are inhib-
itors of cyclooxygenase II (COX-2).
80–82
These substances are
cytotoxic to tumours but not to normal cells.
83,84
An analysis by ow cytometry showed that the caffeic acid
and its derivatives caused cellular entrapment of oral squamous
cell carcinoma cells, Meng-1 (OEC-M1) in the G2/M phase of cell
division. These differential effects on cancer show that these
compounds may be useful in oral cancer chemotherapy.
85
Various natural antioxidants were tested, such as caffeic
acid, catechin, epicatechin and quercetin, to evaluate the
reduction of the cytotoxicity induced by peroxynitrite in chon-
drosarcoma cells. This observation suggests that diets rich in
antioxidant compounds are able to limit cell cytotoxicity.
86
A derivative of caffeic acid, the caffeic acid phenyl ester, was
used and it was observed that it inhibited the growth and
secretion of interleucin-6, and it induced apoptosis in a dose
and time dependent manner on multiple myeloma cells ARH-
77. Therefore, both the caffeic acid and its derivative deserve
further study as effective agents against multiple myeloma.
87
Treatment of mice with propolis, caffeic acid phenyl ester
and caffeic acid itself signicantly reduced the number of
tumour nodules in the lungs of animals whose lung nodules
were generated by injection of viable tumour cells intrave-
nously. In in vitro studies, the propolis did not affect the tumour
cell growth, while the phenyl ester of caffeic acid and caffeic
acid expressed strong cytotoxicity to cells.
88
The p38, a mitogen-activated protein kinase (MAPK),
responds to stimuli such as heat, shock, cytokines, and UV and
is directly involved in cell proliferation and production of NO.
89
Caffeic acid signicantly reduces the mRNA expression of
Interleucin-10 UVB-induced in murine and also inhibits the
activation of p38-MAPK.
90
The caffeic acid also reduces the
migratory capacity of malignant keratinocytes.
91
Although many studies demonstrate the antitumoral activity
of caffeic acid, others show opposite effects. The carcinogenicity
of low dietary levels of the antioxidants caffeic acid, butylated
hydroxyanisole (BHA), sesamol, 4-methoxyphenol (4-MP) and
catechol was examined alone or in combination in a 2 year long-
term experiment. The results indicate that even at low dose
levels phenolic compounds can exert additive/synergistic effects
on carcinogenesis.
92
Furthermore, caffeic acid is also listed
under some Hazard Data and the International Agency for
Research on Cancer has classied it in group 2B as a substance
that is “possibly carcinogenic to humans”.
As there are many controversial results about the protective
or carcinogenic potential of caffeic acid, more studies should be
conducted to elucidate its therapeutic use.
Analytical methods
There is no method for determination and quantication of
caffeic acid in official compendia, such as pharmacopoeias,
however most current articles recommend quantication of
the substance by High Performance Liquid Chromatography
(HPLC).
For determination of rosmarinic and caffeic acids in various
herbs such as rosemary, sage, thyme, mint, lemon balm and
lavender, a HPLC method was developed and validated. The
separation system consisted of a reversed phase C18 column, a
gradient elution system of methanol–water containing phos-
phoric acid and a photodiode array detector. The method
proved to be simple, sensitive, reproducible, fast and ideal for
routine analysis.
93
For quantitation of receptor solution in a permeation study,
the chromatography was performed on a Hypersil ODS column
(particle size: 5 mm25 cm 4.0 mm). The mobile phase was
acetonitrile–water (18 : 82) containing 2% acetic acid. The ow
rate was set at 1.0 mL min
1
. Each sample was ltered before
injection using a Millex lter and an aliquot (20 mL) was
injected onto the HPLC column. Detection was performed at
302 nm.
63
Marti-Mestres and co-workers (2007) conducted a perme-
ation study by applying 50 mL of test formulation 2% caffeic acid
in propyleneglycol–transcutol (1 : 1) on the skin of the pig's ear
and used isotonic saline (NaCl 0.9%) with 1% gentamycin pH
7.0 as the receiver solution. This solution was maintained at
37 C and under these conditions the skin temperature is 32 C,
which corresponds to the temperature of the body surface in
vivo. The quantication of the receptor solution was also carried
out by HPLC with UV detection, column C-8 reverse phase
(5 mm, 250 mm 3 mm) at 40 C. The mobile phase was
acetonitrile–water (18 : 80) with 2% acetic acid and a ow rate of
0.5 mL min
1
. Detection was at 330 nm and the retention time
was approximately 6.0 min.
62
Chromatographic techniques are widely used in the separa-
tion, purication, identication and quantication of
substances. Although HPLC is a robust method, it is more
suitable for mixtures with known compositions. For identica-
tion of unknown substances in complex samples the most
suitable method is MS or LC-MS/MS. Many papers have been
published in recent years on the examination of caffeic acid by
LC-MS/MS approaches.
An example of identication of caffeic acid in a complex
matrix was done for simultaneous analysis of alkamides and
caffeic acid derivatives from Echinacea purpurea extracts. The
analysis was carried out with reversed phase HPLC coupled to
electrospray ionization mass spectrometry (ESI-MS). The ow
rate was set to 0.2 mL min
1
and the eluents were A ¼water +
1% acetic acid and B ¼acetonitrile. The gradient conditions
were: 90% of A in B for 4 min; 90–60% of A in B from 4 to
15 min; 60–40% of A in B from 15 to 30 min; 100% B from 30 to
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35 min; 90% of A in B from 35 to 43 min. The outlet of the HPLC
column was directly connected to the electrospray ionization
source of an ion trap mass spectrometer. The mass spectrom-
eter was operated in the negative ion mode for the rst 15 min
of the analysis, then switched to the positive ion mode for the
remainder. The total analysis time was 43 min. This new
method was considered effective for the quality control of these
extracts that require rapid methods to determine their chemical
composition.
94
Another method was developed and validated to assess the
qualitative and quantitative proles of Myrcia bella hydro-
alcoholic extract. In total, 24 constituents were characterized,
including phenolic acids such as caffeic acid, by means of
extensive preparative chromatographic analyses, along with
mass spectroscopy and Nuclear Magnetic Resonance (NMR)
techniques. And it shows that the mass spectroscopy technique
is a powerful tool for direct and rapid identication of the
constituents aer isolation and NMR characterization.
95
Plasma is another type of complex matrix in which caffeic
acid may be found. A validated method was developed for the
simultaneous determination of the hydroxycinnamates caffeic
acid, dihydrocaffeic acid, ferulic acid, dihydroferulic acid, and
isoferulic acid in human plasma as metabolites derived from
coffee consumption. It was possible using high-performance
liquid chromatography coupled to negative electrospray ioni-
zation tandem mass spectrometry.
96
These same techniques
were used in conjunction to develop a simple, rapid and
sensitive method for the simultaneous quantication of
chlorogenic acid and caffeic acid in rat plasma.
97
Conclusions
This review was carefully prepared to prove the potential of
caffeic acid used in cosmetics and pharmaceutical preparations
and to encourage more studies to elucidate the activity of this
substance on the human body.
Acknowledgements
We thank FAPESP, CNPq and PADC-FCF-UNESP for their
nancial support.
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