ArticlePDF Available

Abstract and Figures

Aims: The purpose of this study was to investigate the action of Citrus limonum essential oil to control free radical-induced lipid peroxidation and preventing tissue damage in skin. Place and Duration of Study: Department of Internal Medicine (University of Roma “Tor Vergata) and A.R.P.A (Aging Research, Prevention and Therapy Association, www.antiaging.it), between January 2010 and June 2011. Methodology: The essential oil was subjected to GC-MS analysis. The superoxide anion scavenging activity of essential oil was evaluated by the enzymatic hypoxanthine/xanthine oxidase system. The same oil diluted in DMSO or grape-seed oil was spread on the face of human volunteers after UV exposition. A sample of skin lipids was collected and the presence of peroxyl radicals was detected based on the measurement of light emitted (chemiluminescence) when the excited carbonyl and singlet oxygen decay to ground state. Results: Our data demonstrate that the lemon essential oil is more active than -tocopherol against *O2- and peroxide free radical inhibition at 1:100 dilution. A protocol for controlling free radical-induced lipid peroxidation in human skin was thus proposed. Conclusion: The scavenging action of lemon essential oil could have a practical application for treating human skin against oxidative damage.
Content may be subject to copyright.
____________________________________________________________________________________________
*Corresponding author: Email: paola.angelini@unipg.it;
European Journal of Medicinal Plants
3(1): 1-9, 2013
SCIENCEDOMAIN international
www.sciencedomain.org
Antioxidative Action of Citrus limonum
Essential Oil on Skin
G. Bertuzzi1, B. Tirillini2, P. Angelini3* and R. Venanzoni3
1Department of Internal Medicine, University of Roma “Tor Vergata”, Via Montpellier,
1 – 00133 Roma (RM), Italy.
2Department of Biomolecular Science, University of Urbino, Via Bramante, 28 - 61028
Urbino (PU), Italy.
3Department of Applied Biology, University of Perugia, Borgo XX giugno 74 - 06121 Perugia
(PG), Italy.
Authors’ contributions
The work presented here was carried out with the collaboration of all the authors. Authors
GB and BT defined the research theme and designed the methods and experiments,
analyzed the data, interpreted the results and wrote the paper. Author PA was involved in
the writing process of the manuscript, author RV co-designed the experiments, discussed
the analyses, interpretation, and presentation of data. All authors have contributed to, seen
and approved the manuscript.
Received 7th August 2012
Accepted 8th October 2012
Published 4th December 2012
ABSTRACT
Aims: The purpose of this study was to investigate the action of Citrus limonum essential
oil to control free radical-induced lipid peroxidation and preventing tissue damage in skin.
Place and Duration of Study: Department of Internal Medicine (University of Roma “Tor
Vergata) and A.R.P.A (Aging Research, Prevention and Therapy Association, www.anti-
aging.it), between January 2010 and June 2011.
Methodology: The essential oil was subjected to GC-MS analysis. The superoxide anion
scavenging activity of essential oil was evaluated by the enzymatic hypoxanthine/xanthine
oxidase system. The same oil diluted in DMSO or grape-seed oil was spread on the face
of human volunteers after UV exposition. A sample of skin lipids was collected and the
presence of peroxyl radicals was detected based on the measurement of light emitted
(chemiluminescence) when the excited carbonyl and singlet oxygen decay to ground
state.
Research Article
European Journal of Medicinal Plants, 3(1): 1-9, 2013
2
Results: Our data demonstrate that the lemon essential oil is more active than -
tocopherol against *O2-and peroxide free radical inhibition at 1:100 dilution. A protocol for
controlling free radical-induced lipid peroxidation in human skin was thus proposed.
Conclusion: The scavenging action of lemon essential oil could have a practical
application for treating human skin against oxidative damage.
Keywords: Anti-aging; GC-MS; grape seed oil; superoxide anion scavenging.
1. INTRODUCTION
The inhibition of lipid oxidation by essential oils such as Origanum spp., Thymus spp.,
Satureja spp. and Rosmarinus officinalis, have already been reported in literature [1, 2, 3].
All the essential oils studied have shown a strong phenolic profile characterized by the
presence of phenylpropanoids which are believed to be the active component of the
essential oils [4, 5, 6, 7, 8, 9]. Citrus essential oil has also been reported to have
antioxidative activities against linoleic acid oxidation [10] and to both Cu2+-induced and 2, 2’-
azobis (2-aminopropane) hydrochloride-induced oxidation of human low-density lipoprotein
in vitro [11]. Among the compounds tested in Citrus essential oil, γ-terpinene had the
strongest antioxidant effect [11], but no clear relationship could be shown between the
antioxidant activity and the essential oil composition of the extracts [12]. When skin is
exposed to air that is irradiated by ultraviolet (UV) light consisting of UVA (320-400 nm) and
UVB (290-320 nm), reactive oxygen species (ROS) including superoxide anion radical (*O2-),
hydrogen peroxide (H2O2), hydroxyl radical (*OH), singlet oxygen (*O2), lipid peroxides
(LOOH), and their radicals (LOO*) are formed. These in turn induce skin aging, phototoxicity,
inflammation and malignant tumors [13, 14, 15, 16, 17].
Recently, consumer interest and the media have focused specifically on products that use
natural ingredients, such as plant extract. There is some evidence that these ingredients
could have possible in vitro anti-aging activity, but the question remains whether it is
possible to deliver adequate doses to the skin in vivo. Lemon oil, traditionally used for its
aromatic properties, has recently been investigated for its effects on skin [18]. The purpose
of this study was to investigate the effectiveness of Citrus limonum Risso essential oil in
controlling free radical-induced lipid peroxidation and preventing tissue damage in skin.
2. MATERIALS AND METHODS
2.1 Plant Material
The lemon (Citrus limonum, Rutaceae) essential oil used in this study was obtained from
cold pressed oil extracted from the peel of the fruit (collected in the north of Sicily Island)
according to the methods of Sawamura and Kuriyama [19]. The cold pressed oil was then
hydrodistilled for 1h in an all-glass Clevenger apparatus. Voucher specimens of C. limonum
plants, identified following the Italian botanical standard treatise [20] were deposited in the
Herbarium of the Dept. of Applied Biology (University of Perugia, Italy).
European Journal of Medicinal Plants, 3(1): 1-9, 2013
3
2.2 GC and GC-MS Analysis
The GC analyses were carried out using a Varian 3300 instrument equipped with an FID and
an HP-InnoWax capillary column (30 m x 0.25 mm, film thickness 0.17 µm), working from
60ºC (3 min) to 210ºC (15 min) at 4ºC/min or an HP-5 capillary column (30 m x 0.25 mm,
film thickness 0.25 µm) working from 60ºC (3 min) to 300ºC (15 min) at 4ºC/min; the injector
and detector temperature was 250ºC. Helium was used as the carrier gas, with a flow rate of
1 ml/min, and the split ratio was 1:10.
GC-MS analyses were carried out with a Hewlett Packard 5890 GC-MS system operating in
the EI mode at 70 eV, using the two above-mentioned columns. The operating conditions
were analogous to those reported in the GC analyses section. The injector and transfer line
temperatures were 220ºC and 280ºC, respectively. Helium was used as the carrier gas, with
a flow rate of 1 ml/min and the split ratio was 1:10.
2.3 Identification of the Components
The components were identified by matching the spectra with those from mass spectral
libraries; the identity of each component was confirmed by comparing the retention indices,
from both columns, relative to the C6-C22 n-alkanes, with those from the literature [21, 22,
23, 24, 25]. When reported,co-elution gas chromatography with reference compounds was
also used for an additional confirmation of the compound identity. The percentage
composition of the essential oil was obtained by the normalization method from the GC peak
areas, without using correction factors.
2.4 Superoxide Anion Scavenging (*O2-)
Superoxide anion was generated by a hypoxanthine-xanthine oxidase system [26]. Reaction
mixtures with 100 µl EDTA (30 mmol/l), 10 µl hypoxanthine (30 mmol/l), 100 µl cytochrome c
(3 mmol/l) or nitroblue tetrazolium (3 mmol/l) were added to 150 µl of lemon essential oil
(solubilized in DMSO 10%) at various concentrations in a final volume of 3 ml buffered in
KH2PO4(50 mmol/l), pH 7.4 [27]. The reaction was started by adding 200 µl xanthine
oxidase (1U/ml) and the rate of reduced cytochrome c or nitroblue tetrazolium was
measured at 550 nm, and 560 nm, respectively, against a reference. The amount of *O2-
generated was calculated using the extinction coefficient ε550 = 2.1 x 10-2 µmol–1 per cm and
the *O2-inhibition was expressed as percentage values. The sample tested did not interfere
with the xanthine oxidase activity (measured at 290 nm). The positive response was tested
using α-tocopherol. Ten repetitions were carried out.
2.5 Randomized Controlled Trial
2.5.1 Subjects
Subjects were selected from among men aged 18 to 52 (mean 33±11) years who were
found to have no serious illness on physical checkup at A.R.P.A. (Aging Research,
Prevention and Therapy Association), www.anti-aging.it (Civita Castellana, VT, Italy). Eighty
volunteers (average age: 33±11 years) who gave their written consent to participate in the
test were selected as subjects from January 2010 to June 2011.
European Journal of Medicinal Plants, 3(1): 1-9, 2013
4
2.5.2 Extraction of skin lipids from healthy volunteers
Skin lipids were collected with acetone-wetted cotton swabs from the forehead over a 9 cm2
area from healthy volunteers (80 men, 18–52 years old–mean 33±11) in the morning for 7
days. The sampling procedure was identical for all the subjects. The volunteers were
randomly divided into four groups (A, B, C, D). In group A the forehead was treated daily for
a week with α-tocopherol in ethanol (20%), group B with lemon essential oil solubilized in
DMSO (1:100), group C with lemon essential oil solubilized in grape-seed oil (1:100), and
group D was left untreated. In accordance with the European norm EN 60335-2-27 and
under medical supervision, volunteers were irradiated daily with UVA and UVB of 0.3
W/m2/mm from sunlamps for 7 min at each session. The participants were asked not to
expose themselves to direct sunlight and to avoid the use of face creams or hair lotions for
the entire duration of the experiment. Twenty-four hours after the last treatment, the skin
lipids were collected.
Extracts were taken twice from the wet cotton swabs using 3 ml of chloroform/methanol
(1:2.5) for two hours (10 µg heneicosanoic acid was used for the recovery test). The raw
extracts were partitioned between 1% NaCl in 0.01 M HCl and chloroform. The chloroform
extracts were washed with methanol/water (1:1) and dried under N2stream. The samples
were stored at –20Cº in 3 ml of chloroform/ethanol (2:1).
2.6 Lipid Peroxidation Analyzed by Chemiluminescence
Chemiluminescence is an index of oxidative stress that quantifies lipid peroxidation and was
measured according to the method of Gonzalez-Flecha et al. [28]. This method is based on
the measurement of light emitted (chemiluminescence) when the excited carbonyl and
singlet oxygen produced by peroxyl radicals decay to ground state. This light is due to the
generation of reactive oxygen species in whole lipids. Skin lipids were incubated with 3 mM
t-BHP for 60 min at 37ºC. Lipid peroxidation was initiated by adding a small amount of stock
solution of t-butyl hydroperoxide (80 mM) to each vial which was then maintained at 37ºC,
and measured by monitoring light emission [29] with a liquid scintillation analyzer Packard
1900 TR. Chemiluminescence was measured over a 60 min period and recorded as counts
per minute (cpm) every 12 min. Each reaction was terminated by adding 5 ml
chloroform/methanol (2:1, v/v) containing 0.01% butylated hydroxytoluene (BHT). This also
inhibited any further oxidation during the lipid extraction. The DMSO had no antioxidative
action and gave a chemiluminescence curve that could be superimposed on to that of the
control.
2.7 Statistical Analysis
Analysis of variance, significances, correlations and other statistical analysis were performed
using Graph Pad Prism version 5.00, (GraphPad Software, San Diego, California, USA).
3. RESULTS AND DISCUSSION
3.1 Chemical Composition of the Essential Oil
Citrus oils are a mixture of volatile compounds and consist mainly of monoterpene
hydrocarbons [30]. Citrus essential oils, on the other hand, generally contain some amount
of coumarins or furanocoumarins [31], flavonoids [32] and tocopherols [33] in the non-volatile
European Journal of Medicinal Plants, 3(1): 1-9, 2013
5
fractions of citrus oils. Coumarins and furanocoumarins may have an important role in skin
photosensitization. Hydrodistillation of the cold pressed oil prevents this hazard. Nineteen
compounds were identified in the GC and GC/MS analyses. The percentage composition of
Citrus limonum essential oil is shown in Table 1. The components are listed in the order of
elution from the HP-5 column. The main component was limonene (54.6%) followed by γ-
terpinene (19.1%) and β-pinene (14.5%). The monoterpene hydrocarbons (87.7%)
constituted the main fraction of lemon oil. This oil composition, as reported in the literature, is
similar to other volatile fractions characterized by the high content of limonene [34].
Table 1. Percentage composition of the essential oil from C. limonum
Compound
%
α-pinene
3,9
β-pinene
14,5
myrcene
1,5
α-terpinene
0,3
p-cymene
0,1
limonene
54,6
γ-terpinene
19,1
terpinolene
0,8
linalool
0,1
citronellal
0,1
terpinen-4-ol
0,1
α-terpineol
0,3
citronellol
0,1
nerol
0,1
neral
1,1
geraniol
0,1
linalyl acetate
0,1
geranial
2,3
geranyl acetate
0,8
aRetention index, relative to C9-C22 n-alkanes on the HP-5 column.
3.2 In Vitro and In Vivo Free Radical Scavenging Activity of Essential Oil
The superoxide anion scavenging activity of Citrus limonum essential oils was evaluated
using the enzymatic hypoxanthine/xanthine oxidase system. Among the concentrations
tested (Fig.1), the 1:100 dilution of lemon essential oil in DMSO had an *O2-inhibition that
was comparable to that of α-tocopherol. The 1:1000 dilution inhibited *O2-less than α-
tocopherol but the level of inhibition was about 76% and 65% of the α-tocopherol activity on
cytochrome c and tetrazolium nitroblue, respectively.
European Journal of Medicinal Plants, 3(1): 1-9, 2013
6
Fig. 1. Percentage cytochrome c inhibition and percentage nitroblue tetrazolium
reduction
Test significant from normal control (P < 0.05). Mean ±S.E.M of ten experiments
The peroxidation data as evidenced by the light emission are shown in Fig 2.
Fig. 2. Chemiluminescence over time in four groups of volunteers
-tocopherol = group A; EO-DMSO = group B ; EO-grape-seed = group C;control = group D.
Test significant from normal control (P < 0.05). Mean ± S.E.M of twenty experiments
The lipids from untreated volunteers showed the highest chemiluminescence and are
considered to be the normal response to the peroxyl radical action. Lower emissions were
recorded for the lipids from volunteers treated with antioxidative substances and the lemon
essential oil was more effective than α-tocopherol as an antioxidant. The grape-seed oil
showed a slightly higher antioxidative action that was added to the action of lemon essential
European Journal of Medicinal Plants, 3(1): 1-9, 2013
6
Fig. 1. Percentage cytochrome c inhibition and percentage nitroblue tetrazolium
reduction
Test significant from normal control (P < 0.05). Mean ±S.E.M of ten experiments
The peroxidation data as evidenced by the light emission are shown in Fig 2.
Fig. 2. Chemiluminescence over time in four groups of volunteers
-tocopherol = group A; EO-DMSO = group B ; EO-grape-seed = group C;control = group D.
Test significant from normal control (P < 0.05). Mean ± S.E.M of twenty experiments
The lipids from untreated volunteers showed the highest chemiluminescence and are
considered to be the normal response to the peroxyl radical action. Lower emissions were
recorded for the lipids from volunteers treated with antioxidative substances and the lemon
essential oil was more effective than α-tocopherol as an antioxidant. The grape-seed oil
showed a slightly higher antioxidative action that was added to the action of lemon essential
European Journal of Medicinal Plants, 3(1): 1-9, 2013
6
Fig. 1. Percentage cytochrome c inhibition and percentage nitroblue tetrazolium
reduction
Test significant from normal control (P < 0.05). Mean ±S.E.M of ten experiments
The peroxidation data as evidenced by the light emission are shown in Fig 2.
Fig. 2. Chemiluminescence over time in four groups of volunteers
-tocopherol = group A; EO-DMSO = group B ; EO-grape-seed = group C;control = group D.
Test significant from normal control (P < 0.05). Mean ± S.E.M of twenty experiments
The lipids from untreated volunteers showed the highest chemiluminescence and are
considered to be the normal response to the peroxyl radical action. Lower emissions were
recorded for the lipids from volunteers treated with antioxidative substances and the lemon
essential oil was more effective than α-tocopherol as an antioxidant. The grape-seed oil
showed a slightly higher antioxidative action that was added to the action of lemon essential
European Journal of Medicinal Plants, 3(1): 1-9, 2013
7
oil; the chemiluminescence curve is a little lower than that of the lemon essential oil
dissolved in DMSO, but the data belong to the same set according to the one-way ANOVA.
These results show that these two oils had a similar scavenging action against peroxide free
radicals in vitro and in vivo [35].
The exposure of human skin to UV radiation can generate ROS in both the epidermis and
dermis. The depth of penetration of UV radiation, as well as its damaging potential in deeper
skin cells have been demonstrated [36]. Among the scavenging substances, α-tocopherol
was chosen as a reference for comparing the scavenging action of lemon essential oil. The
anti-oxidant activity of oil-in-water emulsion containing α-tocopherol has been reported over
a wide range of conditions and test systems [37]. Our data demonstrate that the lemon
essential oil is more active than α-tocopherol against *O2-and peroxide free-radical inhibition
at 1:100 dilution. Lemon essential oil is used instead of other lemon extracts, to avoid the
toxic action that furanocoumarins have under UV exposure.
4. CONCLUSIONS
The results of this study suggest that lemon essential oil has properties that could benefit
human skin as it undergoes environmental and chronological ageing. The scavenging action
of lemon essential oil solubilized in grape-seed oil could have a practical application in
aesthetic medicine (a branch of medicine focused on satisfying the aesthetic desires and
goals of patients) for treating human skin against oxidative damage. Therefore, continuous
application of lemon essential oil solubilized in grape-seed oil might contribute to the
prevention of lifestyle-related skin diseases by regulating the balance of oxidative stress.
ETHICAL APPROVAL
All authors hereby declare that all experiments have been examined and approved by the
appropriate ethics committee and have therefore been performed in accordance with the
ethical standards laid down in the 1964 Declaration of Helsinki.
COMPETING INTERESTS
The authors declare that no competing interests exist.
REFERENCES
1. Estevez M, Cava R. Effectiveness of rosemary essential oil as an inhibitor of lipid and
protein oxidation: contradictory effects in different types of frankfurters. Meat Sci.
2006;72(2):348-355.
2. Kulisic T, Radonic A, Milos M. Inhibition of lard oxidation by fractions of different
essential oils. Grasas y Aceites (Sevilla, Spain). 2005;56(4):284-291.
3. Nakatsu T, Lupo AT, Chinn JW, Kang RKL. Biological activity of essential oils and
their constituents. Stud Nat Prod Chem. 2000;21(B):571-631.
4. Teissedre PL, Waterhouse AL. Inhibition of oxidation of human low-density
lipoproteins by phenolic substances in different essential oils varieties. J Agric Food
Chem. 2000;48(9):3801-3805.
5. Angelini P, Pagiotti R, Menghini A, Vianello B. A antimicrobial activities of various
essential oils against foodborne pathogenic or spoilage moulds. Ann Microb.
2006;56(1):65-69.
European Journal of Medicinal Plants, 3(1): 1-9, 2013
8
6. Angelini P, Granetti B, Pagiotti R. Effect of antimicrobial activity of Melaleuca
alternifolia essential oil on antagonistic potential of Pleurotus species against
Trichoderma harzianum in dual culture. World J Microb Biotech. 2008;24(2):197-202.
7. Angelini P, Pagiotti R, Venanzoni R, Granetti B. Antifungal and allelopathic effects of
Asafoetida against Trichoderma harzianum and Pleurotus spp. Allelopathy J.
2009;23(2):357-368.
8. Pagiotti R, Angelini P, Rubini A, Tirillini B, Granetti B, Venanzoni R. Identification and
characterisation of human pathogenic filamentous fungi and susceptibility to Thymus
schimperi essential oil. Mycoses 2011;54(5):e364-76.
9. Tirillini B, Pagiotti R, Angelini P, Pintore G, Chessa M, Menghini L. Chemical
composition and fungicidal activity of the essential oil of Laserpitium garganicum from
Italy. Chem Nat Comp. 2009;45(1):103-105.
10. Song H-S, Ukeda H, Sawamura M. Antioxidative activities of citrus peel essential oils
and their components against linoleic acid oxidation. Food Sci Technol Res.
2001;7(1):50-56.
11. Takahashi Y, Inaba N, Kuwahara S, Kuki W. Antioxidative effect of citrus essential oil
components on human low-density lipoprotein in vitro. Biosc Biotech Biochem.
2003;67(1):195-197.
12. Di Vaio C, Graziani G, Gaspari A, Scaglione G, Nocerino S, Ritieni A. Essential oils
content and antioxidant properties of peel ethanol extract in 18 lemon cultivars. Sci
Hortic. 2010;126(1):50–55.
13. Bech-Thomsen N, Wulf HC. Carcinogenic and melanogenic effects of a filtered metal
halide UVA source and a tubular fluorescent UVA tanning source with or without
additional solar-simulated UV radiation in hairless mice. Photochem Photobiol.
1995;62(4):773-779.
14. Kligman AM. Early destructive effect of sunlight on human skin. J Am Med Assoc.
1969;210(13):2377-2380.
15. Oikarinen A, Karvonen J, Uitto J, Hannuksela M. Connective tissue alterations in skin
exposed to natural and therapeutic UV-radiation. Photoderm J. 1985;2(1):15-26.
16. Sakurai H, Yasui H, Yamada Y, Nishimura H, Shigemoto M. Detection of reactive
oxygen species in the skin of live mice and rats exposed to UVA light: a research
review on chemiluminescence and trials for UVA protection. Photochem Photobiol Sci.
2005;4(9):715-720.
17. Watson REB, Griffiths CEM. Pathogenic aspects of cutaneous photoaging. J Cosm
Dermat. 2005;4(4):230-236.
18. Chiu A, Kimball AB. Topical vitamins, minerals and botanical ingredients as
modulators of environmental and chronological skin damage. Br J Dermatol.
2003;149(4):681-691.
19. Sawamura M, Kuriyama T. Quantitative determination of volatile constituents in the
pummelo (Citrus grandis Osbeck forma Tosa-buntan). J Agric Food Chem.
1988;36(3):567–569.
20. Pignatti S. Flora d’Italia. Edizioni Edagricole, Bologna, Italy; 1982.
21. Adams RP. Identification of essential oil components by gas
chromatography/quadrupole mass spectroscopy. Allured Publishing Corporation;
2001.
22. Davies NW. Gas chromatographic retention indices of monoterpenes and
sesquiterpenes on methyl silicone and Carbowax 20M phases. Chromatogr.
1990;503:1-24.
23. Heller SR, Milne GWA. EPA/NIH mass spectral data base U. S. Government Printing
Office; 1983.
European Journal of Medicinal Plants, 3(1): 1-9, 2013
9
24. Jennings WG, Shibamoto T. Qualitative analysis of flavour and fragrance volatiles by
glass capillary gas chromatography. Academic Press; 1980.
25. McLafferty FW, Staufer DB. The Wiley NBS registry of mass spectral data. John Wiley
and Sons; 1989.
26. Aruoma O, Halliwell B, Hoey BM, Butler J. The antioxidant action of N-acetylcysteine:
its reaction with H2O2, OH*, OH*2-, HOCl. FRBM. 1989;6(6):593-597.
27. Gressier B, Cabanis A, Lebegue S, Brunet C, Dine T, Luyckx M, Cazin M, Cazin JC.
Decrease of hypochlorous acid and hydroxyl radical generated by stimulated human
neutrophils: comparison in vitro of some thiol-containing drugs. Methods Find Exp Clin
Pharmacol. 1994;16(1):9-13.
28. Gonzalez-Flecha B, Llessuy SF, Boveris A. Hydroperoxide-initiated
chemiluminescence: an assay for oxidative stress in biopsies of heart, liver and
muscle. Free Radic Biol Med. 1991;10(2):93–100.
29. Wright JR, Rumbaugh RC, Colby HD, Miles PR. The relationship between
chemiluminescence and lipid peroxidation in rat hepatic microsomes. Arch Biochem
Biophys. 1979;192(2):344-351.
30. Dugo P, Mondello L, Cogliandro E, Cavazza A, Dugo G. On the genuineness of citrus
essential oils. Part LIII. Determination of the composition of the oxygen heterocyclic
fraction of lemon essential oils (Citrus limon (L.) Burm. f.) by normal-phase high
performance liquid chromatography. Flavour Fragr J. 1998;13(5):329-334.
31. Dellacassa E, Lorenzo D, Moyna P, Verzera A, Mondello L, Dugo P. Uruguayan
essential oils. Part VI. Composition of lemon oil. Flavour Fragr J. 1997;12(4):247- 255.
32. Miyake Y, Yamamoto K, Osawa T. Isolation of eriocitrin(eriodictyol 7-rutinoside) from
lemon fruit (Citrus limon Burm. f.) and its antioxidative activity. Food Sci Technol Int
Tokyo. 1997;3(1): 84-89.
33. Waters RD, Kesterson JW, Braddock RJ. Method for determining the
-tocopherol
content of citrus essential oils. J Food Sci. 1976;41(2):370-371.
34. Espina L, Somolinos M, Lorán S, Conchello P, Pagán DGR. Chemical composition of
commercial citrus fruit essential oils and evaluation of their antimicrobial activity acting
alone or in combined processes. Food Control. 2011;22:896-902.
35. Ahn HS, Jeon TI, Lee JY, Hwang SG, Lim Y, Park DK. Antioxidative activity of
persimmon and grape seed extract: in vitro and in vivo. Nutr Res. 2002;22(6):1265-
1273.
36. Katiyar SK, Afaq F, Perez A, Mukhtar H. Green tea polyphenol (-)-epigallocatechin-3-
gallate treatment of human skin inhibits ultraviolet radiation-induced oxidative stress. J
Carcinog. 2001;22(2):287-294.
37. Frankel EN, Huang SW, Kanner J, German JB. Interfacial phenomena in the
evaluation of antioxidants: bulk oils vs emulsions. J Agric Food Chem.
1994;42(5):1054-1059.
_________________________________________________________________________
© 2013 Bertuzzi et al.; This is an Open Access article distributed under the terms of the Creative Commons
Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited.
Peer-review history:
The peer review history for this paper can be accessed here:
http://www.sciencedomain.org/review-history.php?iid=159&id=13&aid=729.
... The lemon essential oil extracted from Citrus limonum Risso has properties that can benefit human skin, has a scavenging action, and can be used to heal oxidative damaged human skin in esthetic medicine. In addition, it was reported in the study that the permanent application of lemon essential oil solubilized in grape-seed oil might contribute to the avoidance of lifestyle-related skin illnesses by controlling the oxidative stress level [40]. ...
... Citrus fruits are known to be rich in vitamins C and B and used as a good source of folate, and they are among the dietary fiber sources due to the dietary fibers they contain. The effects on the peel essential oils, pectic polysaccharides (pectin), flavonoids, and organic acids of fruits, as well as on health, are revealed by research, and such studies keep up to date [40][41][42]. In addition, the citrus species' leaves, flowers, peels, and seeds are the focus of the attention of researchers [6]. ...
... Citrus EO components exhibit antioxidative activities against the oxidation of linoleic acid. In addition, antioxidant activities have also been reported against in vitro oxidation of human low-density lipoprotein induced by Cu 2+ , and 2, 2 -azobis (2-aminopropane) hydrochloride [128]. The antioxidant properties of citrus EOs are attributed to the presence of phenolic compounds in their composition. ...
... Scalvenzi et al. [127] reported that the EO of M. splendens showed cytotoxic activity against the human tumor cell lines MCF-7 (breast) and A549 (lung) and the nontumor cell line HaCaT (human keratinocytes). Melo et al. [128] stated that the EO of M. lundiana showed toxicity against the insect Acromyrmex balzani. ...
... Furthermore, they are widely employed as flavoring agents in various food, beverage, and confectionery products [10]. The beneficial properties of citrus EOs encompass a range of advantages, including antioxidant [11,12], antimicrobial [13], antiviral [14], anti-inflammatory [15], and anticancer [16] properties. Given their numerous benefits, EOs find wide applications in the agricultural, food, cosmetic, textile, and pharmaceutical industries [17,18]. ...
Article
Full-text available
Citrus fruits, members of the Rutaceae family, have seen a surge in the popularity of their essential oils (EO) due to their versatile industrial applications. These EOs are primarily derived from citrus fruit peels, a practice that not only reduces waste generation but also minimizes environmental contamination. Citrus EO serves as a natural and cost-effective substitute for synthetic preservatives and flavoring agents, contributing to the pursuit of safe and wholesome food, a paramount goal in the food industry. The antimicrobial properties of key compounds such as D-limonene, linalool, α & β-pinene, sabinene, β-myrcene, α-terpineol, and other bioactive chemicals are well-documented. Moreover, these components exhibit antioxidative and potential anticancer attributes. Additionally, citrus EO-based films and coatings offer diverse applications in the realm of food packaging materials. This comprehensive review delves into a variety of extraction methods, component characterization, and recent applications of citrus essential oils across different food categories. As the demand for safe and natural food continues to grow, citrus essential oils employed as food preservatives hold a promising future. Nonetheless, further research is imperative to explore broader applications and ascertain potential allergenic and toxicological consequences, ensuring the continued advancement of this promising field.
... These substances have the capacity to shield cells and tissue from UV radiation. As a result, lotions or ointments containing these essential oils might make the claim that they shield skin from UV rays and prevent oxidation (Bertuzzi et al. 2013;Wei and Shibamoto 2007). The term "nano-cosmeceuticals" refers to a cosmetic formulation that uses nanotechnology as a means of delivering bioactive ingredients to the skin (Hougeir and Kircik 2012;Kaul et al. 2018). ...
Chapter
Essential oils (EOs) are complex mixtures of volatile secondary metabolites extracted from different parts of aromatic plants such as leaves, flowers, fruits, and seeds. They are also referred to as aromatic oils. In general, EOs are cocktail of different low-molecular-weight compounds, such as flavonoids, terpenoids, terpenes, and phenylpropanoids. They represent a diverse class of stereochemistry that results in a range of medicinal properties, viz., antimicrobial, antioxidant, anti-inflammatory, and antiviral effects. Additionally, EOs are biodegradable and hardly affect non-targeted species, which can be beneficial in delaying the development of resistance. EOs have been used for a long time in various areas such as food, medicine, cosmetics, and aromatherapy. However, due to certain limitations associated with them such as high volatility, intense aroma, and chemotypic variation, etc., they were replaced by chemical alternatives that were more efficient and better in terms of bioactivity. However, in view of green consumerism and the increased negative concerns (non-biodegradability and the adverse effects of their by-products on the environment and human health) associated with the indiscriminate use of synthetic chemicals, industries are looking toward green chemicals as a preferred alternative to synthetic ones.
... They are high in vitamin C, which is an antioxidant and a brightening aging. They remove excess oil from your skin and reduce acne (Bertuzzi et al., 2013). ...
... The major chemical compounds are the limonene (66%), the β-pinene (13.80%) and the γ-terpinene (9, 10%). The previous work has revealed the existence of majority compounds as limonene (54.6%), γ-terpinene (19.1%) and β-pinene (14.5%) [3] There are many factors that can explain the difference in the chemical composition of the EOs of C. limonum between the result of our study and it's of the anterior work such as such as climate regions of origin of the samples, the genetics of the plant, altitude and the soil. ...
Article
Full-text available
The three botanical species: Mentha spicata, Thymus vulgaris, and Citrus limonum are a medicinal plants widely used in Morocco. The aromatic fractions of these plants offer new perspectives in herbal medicine through the development of new pharmaceutical preparations for therapeutic purposes. The essential oils (EOs) are extracted from dried plants, in the open air and away from light. The choice of these medicinal plants was made following an investigation and a statistical study conducted in various regions of Morocco. The results of physico-chemical analysis of the EOs oils are consistent with those of the AFNOR [1] standards. The analysis of their chemical composition was determined by gas chromatography-mass spectrometry (GC/MS). The antibacterial activity of these EOs was tested in two types of bacterial germs and the results for the in vitro activity show that these two germs has shown high sensitivity to these three EOs.
... Citrus EO components exhibit antioxidative activities against the oxidation of linoleic acid. In addition, antioxidant activities have also been reported against in vitro oxidation of human low-density lipoprotein induced by Cu 2+ , and 2, 2′-azobis (2-aminopropane) hydrochloride [128]. The antioxidant properties of citrus EOs are attributed to the presence of phenolic compounds in their composition. ...
Article
Full-text available
Citrus is one of the main fruit crops cultivated in tropical and subtropical regions worldwide. Approximately half (40–47%) of the fruit mass is inedible and discarded as waste after processing, which causes pollution to the environment. Essential oils (EOs) are aromatic compounds found in significant quantities in oil sacs or oil glands present in the leaves, flowers, and fruit peels (mainly the flavedo part). Citrus EO is a complex mixture of ~400 compounds and has been found to be useful in aromatic infusions for personal health care, perfumes, pharmaceuticals, color enhancers in foods and beverages, and aromatherapy. The citrus EOs possess a pleasant scent, and impart relaxing, calming, mood-uplifting, and cheer-enhancing effects. In aromatherapy, it is applied either in message oils or in diffusion sprays for homes and vehicle sittings. The diffusion creates a fresh feeling and enhances relaxation from stress and anxiety and helps uplifting mood and boosting emotional and physical energy. This review presents a comprehensive outlook on the composition, properties, characterization, and mechanism of action of the citrus EOs in various health-related issues, with a focus on its antioxidant properties.
Preprint
Full-text available
The study aimed to explore the potential of lemon peel extracts as a cosmetic raw material, with a focus on their aromatic, antimicrobial and various biological activities. Lemon peel essential oil (PLEO), extract (PLE) and absolute oil (PLAO) were prepared by steam distillation and organic solvent extraction, respectively. The compositions of PLEO, PLE and PLAO were analyzed by GC/MS, revealing 22, 39, and 9 components respectively, with terpenoids being the main component. PLE had the highest total flavonoid content, and surpassing that of total polyphenols. Aroma intensity, measured with an electronic pen, followed the order LPEO > LPE > LPAO, while aroma persistence was ranked LPAO > LPEO > LPE. All three lemon peel extracts showed strong antibacterial (against E. coli , S. aureus , and C.albicans ), as well as antioxidant and anti-tyrosinase properties, with inhibition rates exceeding 90% in a dose-dependent manner. LPEO demonstrated superior anti-inflammatory effects compared to LPAO and LPE, with inhibitory rates of 87.79 ± 3.86% and 80.75 ± 2.33% on TNF-α and IL-6 at a concentration of 1×10 − 2 mg/mL. Lemon peel extract was found to promote HaCat cell migration, with LPEO showing greater effectiveness than LPE and LPAO. The healing rate of scratched HaCat cells treated with LPEO at a concentration of 1×10 − 2 µL/mL for 12 hours was 95.29 ± 3.41%. In conclusion, the combination of these extracts could broaden their applications in cosmetics by offering aroma-enhancing, antioxidant, whitening, antibacterial, anti-inflammatory, and skin wound healing benefits.
Article
Full-text available
Documentation of indigenous knowledge about plants plays a key role for conservation and utilization of plant resources. The present paper documents the diversity of plants and their traditional use in and around the Jhilmil Lake Area, one of the forest-dominated peri-urban areas of Kanchanpur district lying in Sudurpaschim Province. Vegetation sampling and ethnobotanical surveys were carried out twice between January, 2020 and January, 2021. Semi-structured questionnaire and checklists were used to record the use and distribution of the plant species and their conservation. A total of 126 plant species representing 52 families and 113 genera were reported. Among the total plants recorded, 114 (90.48 %) species were found to be ethnomedicinally used. The results showed that the use of plants as ethnomedicine was culturally motivated and less influenced by availability of the plants. The plant “importance value index” (IVI) was found to be negatively associated with the plant “relative frequency of citation” (RFC; p=0.057–0.790). The high RFC values of the trees and climbers hinted that the plant collection was subjectively oriented towards quality products. The findings suggested that the rare plants with high-use values such as Pterocarpus marsupium, Dalbergia latifolia, Rauvolfia serpentina, Citrus limonum, and Mussaenda frondosa should be prioritised for future conservation. It illustrates that lakes in forested areas are essential resources for plant diversity and local life because they include a variety of rare and useful local plant species
Article
Full-text available
Fish by-product oil and lemon oil have potential applications as active ingredients in many industries, including cosmetics, pharmaceuticals and food. However, the physicochemical properties, especially the poor stability, compromised the usage. Generally, nanoemulsions were used as an approach to stabilize the oils. This study employed an ultrasonication method to form oil-in-water nanoemulsion of lemon and fish by-product oils (NE-FLO). The formulation is produced at a fixed amount of 2 wt% fish by-product oil, 8 wt% lemon oil, 10 wt% surfactant, 27.7 wt% co-surfactants and 42 min of ultrasonication time. The size, polydispersity index (PDI) and zeta potential obtained were 44.40 nm, 0.077, and −5.02 mV, respectively. The biological properties, including antioxidant, antibacterial, cell cytotoxicity, and anti-inflammatory, showed outstanding performance. The antioxidant activity is comparable without any significant difference with ascorbic acid as standard and is superior to pure lemon oil. NE-FLO successfully inhibits seven Gram-positive and seven Gram-negative bacterial strains. NE-FLO’s anti-inflammatory activity is 99.72%, comparable to nordihydroguaiaretic acid (NDGA) as the standard. At a high concentration of 10,000 µg. mL−1, NE-FLO is non-toxic to normal skin cells. These findings demonstrate that the NE-FLO produced in this study has significant potential for usage in various industries.
Article
Full-text available
Adams, R. P. 2007. Identification of essential oil components by gas chromatography/ mass spectrometry, 4th Edition. Allured Publ., Carol Stream, IL Is out of print, but you can obtain a free pdf of it at www.juniperus.org
Article
Full-text available
The composition of 31 genuine Uruguayan lemon oils obtained by industrial processing (FMC on line) during the 1995 season is reported. The samples were representative of all production areas situated in the North and South of Uruguay. The volatile fraction was analysed by HRGC–FID and GC–MS; the enantiomeric distribution of β-pinene, sabinene, limonene, linalol, terpinen-4-ol and α-terpineol was studied by multidimensional HRGC–HRGC; coumarins and psoralens present in the non-volatile residue were analysed by normal phase HPLC. The results relative to the volatile fraction were compared with those obtained for Uruguayan oils produced in the 1992 and 1993 seasons and for Italian FMC oils. Coumarins and psoralens have been analysed for the first time in Uruguayan oils; their content is compared with that of Italian oils. © 1997 John Wiley & Sons, Ltd.
Article
Full-text available
Composition of essential oils and antioxidant activity of peel ethanol extract were analyzed in 18 local lemon cultivars. Essential oils composition was determined by GC/FID analysis, and antioxidant activity with the ABTS method. Fruit weight, polar and equatorial diameters, peel thickness, seed number, juice percentage, titratable acidity and juice pH were also determined for each cultivar. The main component in the peel essential oil was limonene, accounting for 72.5–76.4%, followed by β-pinene (11.6–18.7%). Several other monoterpene hydrocarbons were also identified at appreciable contents, namely terpinene (2.88–8.26%), α-pinene (1.4–1.5%) and myrcene (0.95–1.12%). No clear relationship could be shown between the antioxidant activity and the essential oil composition of the extracts. In this study, cultivars with higher essential oil content and antioxidant activity were identified. Data were subjected to analysis of variance using ANOVA and means were compared by the Duncan test.
Article
We determined in vitro radical scavenging activity of persimmon seed extract (PSE) and grape seed extract (GSE), and quantified total tannin concentrations of each extract. It has been found that both PSE and GSE have radical scavenging activities, and total tannin concentration of PSE was significantly higher than GSE (p < 0.05). In order to investigate the protective effect on oxidative stress in the liver, rats were administered PSE and GSE, respectively. PSE significantly decreased the liver thiobarbituric acid reactive substances (TBARS) and phosphafidylcholine hydroperoxides (PCOOH) level compared to control group (p < 0.05). Furthermore, catalase and superoxide dismutase (SOD) activities were increased by both PSE and GSE administration. However, fatty acid composition in phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were not significantly different among control and other group rats. We concluded that PSE and GSE have strong radical scavenging activity in vitro, and inhibited lipid peroxidation in vivo.
Article
An antioxidant was isolated from the peel and juice of lemon fruit (Citrus limon BURM. f.). It was identified as eriocitrin (eriodictyol 7-rutinoside) of the flavanone glycoside by HPLC, 1H-NMR and 13C-NMR analyses. The purified eriocitrin was readily soluble in water, methanol, and ethanol. A water solution of 0.05% eriocitrin was weakly acidic (pH 4.2). Eriocitrin was found to be stable even at high temperature (121°C, 15 min) in acidic solution (pH 3.5). The distribution of eriocitrin in citrus fruits was found to be especially abundant in lemons and limes, however, it was scarcely found in other citrus fruits. In the case of lemon fruit, eriocitrin was primarily distributed in the peel (about 1,500 ppm) composed of the albedo (mesocarp), flavedo (epicarp), and pulp vesicles. It was also significantly present in the juice (about 200 ppm) but was not detected in the seed. Two varieties of lemon fruits, eureka and lisbon, almost had the same eriocitrin content. The antioxidative activity of eriocitrin in the linoleic acid autoxidation system was equal to that of α-tocopherol, and it was enhanced when used together with citric acid. The eriocitrin had a synergistic effect on α-tocopherol.
Article
Antioxidative activities of thirty kinds of citrus essential oils and their fourteen components against linoleic acid oxidation were examined by a thiocyanate method. The tocopherol contents of these oils were determined by high-performance liquid chromatography monitored at 290 nm. All the citrus essential oils and the flavor components showed inhibitory effects against linoleic acid oxidation. Highest antioxidative ability was observed in the essential oils of yuzu, lemon, hassaku, sudachi, mochiyu, yuko and Tarocco orange. Antioxidative activities of β-pinene, myrcene, α-terpinene, γ-terpinene and decanal were higher or similar to that of δ-tocopherol. Tocopherols were contained abundantly in the essential oils of limes, Valencia orange, yuzu and lemons. However, there was little correlation between tocopherol contents and antioxidative activities in citrus essential oils. The composition of terpene compounds seems to be a major factor in the antioxidative activities of these oils.
Article
This Supplement to the EPA/NIH Mass Spectral Data Base (NSRDS-NBS 63) presents an additional collection of 8807 verified mass spectra of individual substances compiled from the EPA/NIH mass spectral file. The spectra are given in bar graph format over the full mass range. Each spectrum is accompanied by a Chemical Abstracts Index substance name, molecular formula, molecular weight, structural formula, and Chemical Abstracts Service Registry Number. A cumulative index has also been issued which provides access to the entire file of 34,363 mass spectra. (Author)