A comprehensive review on Citrus aurantifolia essential oil: its phytochemistry and pharmacological aspects

Abstract

Citrus essential oil, commonly, known as lime oil, has been widely reported in traditional system of medicine. Industrially, oil is isolated by mainly by hydrodistillation from fruit and peel of Citrus aurantifolia, family, Rutaceae. Cultivation practice of citrus plants dates back for over 4000 years and are one of most valuable fruit crops in the world. In this review, we aim to summarise the phytochemical and biological properties of citrus oil. The literature was collected from various online resources such as e journals, books and magazines. The citrus essential oil is globally used in food industry to impart citric flavour and odour to cuisines. Llime juice and oil is known to possess multiple biological properties including anti-cancer, antimicrobial, antioxidant, antiulcer, anti-inflammatory, hypolipidemic, antityphoid and hepatoprotective properties. Due to potent antibacterial and antifungal properties, citrus oil is becoming important component of skin care products. The medicinal importance of plant is due to presence of various secondary metabolites, alkaloids, carotenoids, coumarins, essential oils, flavonoids, phenolic acids, and triterpenoids. The citrus oil is rich in aromatic compounds namely, monoterpenes and their derivatives, aldehydes, ketones, esters, alcohols such as limonene (58.4%), β-pinene (15.4%), γ-terpinene (8.5%), citral (4.4%) and others. The bitter taste and aroma of citrus fruit peels is attributed to limonoids. p-caryophyllene constitute 5.7% of all the sesquiterpenes. On the basis of the available information, we conclude that citrus oil possess huge potential to be developed into pharmaceutical products.

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ISSN 2595-0584 - V.3 N.2
July 2020 - pag. 354 - 364
Electronic journal
Brazilian Journal of Natural Sciences www.bjns.com.br
Brazilian Journal of Natural Sciences
¹Department of Pharmacognosy and Phytochemistry, School of Pharmaceutical Sciences, Delhi Pharmaceutical Sciences
and Research University, New Delhi, India. ²Department of Pharmaceutics, School of Pharmaceutical Sciences, Delhi
Pharmaceutical Sciences and Research University, New Delhi, India.
Authors: Shagun Jain¹, Poonam Arora1,A, Harvinder Popli2,B
A COMPREHENSIVE REVIEW ON CITRUS AURANTIFOLIA ESSENTIAL
OIL: ITS PHYTOCHEMISTRY AND PHARMACOLOGICAL ASPECTS
Review Article
ACorresponding author:
Poonam Arora, Associate Professor (Temporary faculty), Department of Pharmacognosy and Phytochemistry, School
of Pharmaceutical Sciences, Delhi Pharmaceutical Sciences and Research University, New Delhi, India. E-mail:
poonamarora96@gmail.com - https://orcid.org/0000-0002-4118-8180. Contact no.: 91-9811551378.
DOI: https://doi.org/10.31415/bjns.v3i2.101 - Article received on July 01 , 2020; Accepted on July 07, 2020; published
on 23 July 2020 in the Brazilian Journal of Natural Sciences, Vol. 3, N.2, online, ISSN 2595–0584. www.bjns.com.br. All
authors contributed equally to the article. The authors declare that there is no conict of interest. This is an open access
article under the CC - BY license: http://creativecommons.org/licenses/by/4.0.
Abstract
Citrus essential oil, commonly, known as lime oil, has been widely
reported in traditional system of medicine. Industrially, oil is isolated by
mainly by hydrodistillation from fruit and peel of Citrus aurantifolia,
family, Rutaceae. Cultivation practice of citrus plants dates back for over
4000 years and are one of most valuable fruit crops in the world. In this
review, we aim to summarise the phytochemical and biological properties
of citrus oil. e literature was collected from various online resources
such as e journals, books and magazines. e citrus essential oil is globally
used in food industry to impart citric avour and odour to cuisines. Llime
juice and oil is known to possess multiple biological properties including
anti-cancer, antimicrobial, antioxidant, antiulcer, anti-inammatory,
hypolipidemic, antityphoid and hepatoprotective properties. Due to potent
antibacterial and antifungal properties, citrus oil is becoming important
component of skin care products. e medicinal importance of plant is
due to presence of various secondary metabolites, alkaloids, carotenoids,
coumarins, essential oils, avonoids, phenolic acids, and triterpenoids.
e citrus oil is rich in aromatic compounds namely, monoterpenes and
Key words:
Citrus essential oil,
monoterpenes, limonene,
antifungal, antioxidant.
Article ID
BCorresponding author:
Harvinder Popli, Dean and Professor, Department of Pharmaceutics, School of Pharmaceutical Sciences, Delhi
Pharmaceutical Sciences and Research University, New Delhi, India. E. mail: popli.harvinder@gmail.com - https://orcid.
org/0000-0001-9030-5462. Contact no.: 91-98997 00704.

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Introduction
e genus Citrus (Rutaceae) is one of the most
widely consumed and economic important group
(1). e global production of citrus fruits has
signicantly increased to 82 million tons in the
years 2009–2010 (2). Around 70% of the world’s
total marketable citrus are grown in the America,
Brazil, Mediterranean countries. Of these, India is
the world’s largest producer of dierent varities of
limes (Table 1) while China produces most of the
world’s mandarins an important variety of lime
(3,4). Citrus products are a rich source of vitamins,
minerals and dietary bers that are essential for
growth and development of body. e fruits possess
their derivatives, aldehydes, ketones, esters, alcohols such as limonene
(58.4%), β-pinene (15.4%), γ-terpinene (8.5%), citral (4.4%) and others.
e bitter taste and aroma of citrus fruit peels is attributed to limonoids.
p-caryophyllene constitute 5.7% of all the sesquiterpenes. On the basis of the
available information, we conclude that citrus oil possess huge potential to
be developed into pharmaceutical products.
anti-cancer (5), antimicrobial, antioxidant (6),
antiulcer, anti-inammatory, and hypolipidemic,
antityphoid and hepatoprotective (7) properties.
e essential oils contain many volatile compounds,
mainly aldehydes, ketones, esters, alcohols and
terpenes, which give the characteristic aromas
and avours of the citrus fruits. Because of their
nutritional values and pleasant aromas, some of
citrus fruit juices are used as functional drink (3, 8).
Limonoids are the principal compounds found in
citrus fruit peels where they produce the bitter taste
and the zest aroma (9–11). Citrus-peel essential oils
are amongst the most important of these, including
orange, lemon, mandarin, tangerine and grapefruit
oils are leading oils in terms of volume (12,13).
Table 1. Some popular varieties of citrus
S. No. Var i et y Reference
1 Sweet oranges (Citrus sinensis Osbeck) (14)
2Mandarins (Citrus reticulata Blanco) (15,16)
3 Grapefruits (Citrus paradisi Macfadyen) (17)
4 Lemons (Citrus limon Burmann) (18)
5Limes (Citrus aurantifolia Swingle) (19)
Chemical constituents
e peculiar phytochemical composition of the
peel and leaf oils of C. aurantifolia suggest use of the
essential oils as a characteristic taxonomic marker
for species (20) (Table 2, Fig. 1). e phytochemistry
of citrus oil has been studied extensively by many
researchers. GC-FID and GC-MS of hydrodistilled
essential oil of C. aurantifolia, shows presence of
limonene (58.4%), β- pinene (15.4%), β-terpinene
(8.5%), and citral (4.4%) as the major constituents (21).
Some exclusive terpenes such as the sesquiterpene
santal-10-en-2-ol have been identied in the lime
peel oil (22). An oxygenated monoterpene, fenchol,
has also been isolated in C. aurantifolia (23,24).
Some othermono- and sesquiterpene hydrocarbons
and oxygenated monoterpenes such as β-pinene,
neryl acetate, geranyl acetate, β-bisabolene, (E)-α-
bergamotene, germacrene D and β-caryophyllene
(25) have also been reported in C. aurantifolia. In
addition, lime oil also contain coumarins which are
known to cause phototoxic reaction in humans. In
experimental animals, these coumarins were found
to promote tumour formation on skin and abdominal
epithelium of mice induced by 9,10-dimethyl-1,2-
benxanthracene and benzo-[a]-pyrene (26–28).

356 ARORA, P. et al - Braz. J. Nat. Sci. – electronic journal ISSN: 2595-0584 - V.3 - N.2
C. aurantifolia peel oil
e chemical composition of C. aurantifolia peel
oil is very similar to that of C. hystrix, a Malaysian
citrus species with presence of monoterpenes (94.6%).
e two most abundant compounds were limonene
(39.3%) and p-pinene (28.4%). However, the former
can be distinguished by the presence of relatively
high concentrations of geraniol (7.5%), neral (5.3%)
and geranial (2.1%), citronellal (0.1%) with absolute
absence of citronellol. GC-MS analysis of some species
of citrus, C. hystrix D.C., C. aurantifolia Swingle, C.
maxim Merr. and C. microcarpa Bunge, revealed that
C. hystrix peel oil comprises mainly of monoterpenes
(97.2%) with p-pinene (39.3%), limonene (14.2%),
citronellal (11.7%) and terpinen-4-ol (8.9%) as the
major components. Other monoterpenes present
in appreciable amounts include α-terpineol (3.0%),
terpinene (2.4%), α-pinene (2.0%), linalool (l.9%) and
furanoid cis-linalool oxide (1.9%). 17 sesquiterpenoids
in smsll quantities constituting 2.6% of the oil have
also been identied in the lime essential oil. Myrcene,
is present at 1.6% and 1.8% concentrations in the peel
oils of C. maxima and C. microcarpa. In comparison,
peel oils of C. maxima and C. microcarpa contained
more than 94% of monoterpene hydrocarbon,
limonene, and could be one of the important natural
sources of limonene.
C. aurantifolia leaf oil
e leaf oil of C. aurantifolia contain the highest
concentration of monoterpenes amongst other species
of citrus. Geranial (19.4%), limonene (16.4%), neral
(11.4%), nerol (9.5%), geraniol (7.5%) and geranyl
acetate (6.6%) are the major constituents of the leaf oil
of C. aurantifolia. Sesquiterpenes present in amounts
greater than 1% concentration are p-caryophyllene
(5.7%), (Z)-nerolidol (2.0%), (Z)-p-farnesene (1.8%)
and p-elemene (1.6%). whilst, the leaf oil of C. hystrix
contains mainly citronellal (72.4%) and related
compounds, citronellol (6.7%) and citronellyl acetate
(4.1%). Of the other 39 components present in the leaf
oil of C. hystrix, only p-pinene (1.9%), linalool (l.7%)
and trans-sabinene hydrate (1.5%) are present at greater
than 1% concentration. Sesquiterpenes accounts for
only 4.5% of the oil. In contrast, C. macrocarpa leaf
oil possess more than 70.8% sesquiterpenes with
hedycaryol (19.0%), p-sesquiphellandrene (18.3%),
α-eudesmol (l4.4%) and p-eudesmol (8.6%). While,
p-pinene (13.4%), linalool (6.1%) and (E)-p-ocimene
(2.0%) are main the monoterpenes C. macrocarpa leaf.
Table 2. List of phytochemicals in C. aurantifolia
S. No. Type Compounds Reference
1Sugars Glucose, fructose and sucrose (1-15%) (10,29,30)
2 Polysaccharides Cellulose, hemicelluloses and pectin (31)
3 Organic acids
Citric and malic acids with small quantities of succinic,
malonic, lactic, oxalic, phosphoric, tartaric, adipic and
isocitric acids
(20,32)
4Lipids Phospholipids (0.1%), palmitic, palmitoleic, oleic,
linoleic and linolenic acids (14,33)
5 vitamins
Ascorbic acid, thiamine, riboavin, niacin, pantothenic
acid, inositol, biotin, vitamin A, vitamin K, pyridoxine,
paminobenzoic acid, choline and folic acid
(33)
6 Inorganic elements Potassium and nitrogen (80%), calcium, iron,
phosphorus, magnesium and chlorine (34)
7Flavonoids Flavanones, avones and anthocyanins (1,31,35)
8Limonoids Limonene (36,37)
9Volatile compound Limonene (21,38)

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ARORA, P. et al - Braz. J. Nat. Sci. – electronic journal ISSN: 2595-0584 - V.3 - N.2
Fig. 1. Structures of major chemical constituents present in C. aurantifolia essential oil
Health Benets of phytochemicals isolated from
Citrus
Citrus is rich in avonoids including apigenin,
rutin, quercetin, kaempferol, nobiletin, hesperidin,
hesperitin, and neohesperidin. Quercetin, has been
reported as one of the most active avonoids that
possess signicant anti-inammatory, anti-tumor,
anticancer, anti-prostatitis, anti-allergic and anti-
asthmatic (31,39-43). Carotenoids found in citrus
are β-carotene, lutein, zeaxanthin and cryptoxanthin
(44,45). Presence of vitamin C in citrus enhances
its medical applicability in treatment of stress, cold,
chills, muscle fatigue and scurvy (40, 46–50).
Pharmacological activities of C. aurantifolia
Pharmacological activities of the extract of
dierent parts of C. aurantifolia have been studied.
e plant possesses the numerous biological activities
described below:
Antibacterial activity
Antimicrobial activity of citrus oil against several
pathogens, including, S. aureus, Escherichia coli,

358 ARORA, P. et al - Braz. J. Nat. Sci. – electronic journal ISSN: 2595-0584 - V.3 - N.2
Klebsiella pneumonia, Pseudomonas spp, A. niger and
C. albicans, has extensively been studied (28,51–55).
Hydrodistilled lime oil (12.25-100 μg/ml) possess
potent antibacterial activity against gram positive
compared to gram negative strains (56). e values of
zone of inhibition (ZOI) recorded for lime essential
oil against some microbes investigated in study
were, S. aureus (10 to 20mm), Enterococcus faecalis
(26mm), Salmonella spp. (6-10mm) and C.albicans
(24 mm). e oil demonstrates powerful results
in isoniazid-resistant strain of Mycobacteria that
suggest probably, oil could have role in overcoming
antimicrobial resistance (57,58). e antibacterial
activity of C. aurantifolia has been attributed to the
presence of phytochemicals, 5, 8-dimethoxypsoralen,
5-geranyloxypsoralen, palmitic acid, linoleic acid,
oleic acid, 4-hexan-3-one and citral (58).
Abubakar U Zage found that citrus ethanolic
extract (2.125-20 mg/ml) shows signicant activity
against clinical isolates of Shigella, Salmonella typhi,
Klebsiella. e study indicated that Shigella was more
sensitive to the extract with average zone of inhibition
of 14.90 mm, followed by Klebsiella (14.49 mm), E.
coli (13.77 mm) and S. typhi (12.01 mm) (59–61).
In comparison, citrus peel methanolic extract is
potent against S. aureus at concentration 31.25 µg/
ml, while ethyl acetate extract is eective at higher
concentration, 250-750 µg/ml (62).
Antifungal
e oil is becoming an important component of
dermatological formulations used in skin and scalp
diseases (63). Antifungal eects of citrus oil has been
studied against Malassezia furfur in in vitro model
using disk diusion method. In a study, oil elicited
fungistatic eects in a dose dependent manner with
zone of inhibition of 2.6 mm at minimum inhibitory
concentration (MIC) 2 mg/ml when compared to oil-
untreated culture (64,65). Lime essential oil at 2 mg/
ml was found to inhibit growth of M. furfur, KCCM
12679 cultured on sabrouds dextrose agar media with
incubation temperature 37°C for 2-7 days, ZOI was
found to be 2.6 mm when compared with reference
standard (66).
In some studies, A. niger have shown explicitly
high susceptibility to oil isolated from lime leaves (55).
Matan. N. concluded that limonene in lime oil inhibits
the growth of A. niger cultured in potato dextrose
agar medium at 70°C. Additionally, monoterpene
hydrocarbon, at MIC 90 µl/ml, also showed synergistic
activity with other secondary metabolites present in
lime oil (67, 68). Due to antifungal activity of citrus,
the plant may be a potential candidate for use in
agriculture and food industry for protection against
aatoxin contamination. e eects of lime essential
oil on some species of molds has been determined by
Matan and Matan. At concentration, 20–200 µl/ml,
the oil was found to be fungistatic as well as fungicidal
on the test species. Lime oil was eective at 100 µl/ml
against P. chrysogenum and Penicillium sp. while A.
niger was susceptibile at only higher concentrations of
lime oil (140 µl/ml). e MIC values performed by the
broth dilution of all conditions were examined (70).
Anti-obesity activity
Co-administration of C. aurantifolia essential oil
with ketotifen in wistar mice suppressed weight gain
in animals. e weight loss was described due to
possibility of promoting anorexia, reduction in both
the amount of food intake compared with the control
group. (71)
Anticancer/cytotoxic activity
C. aurantifolia fruit from Texas, USA, consists of at
least 22 volatile compounds, and its major compounds
limonene (30%) and dihydrocarvone (31%) and
ve active components of C. aurantifolia seeds
such as limonin, limonexic acid, isolimonexic acid,
β-sitosterol glucoside, and limonin glucoside. Patil
and group reported that 100 µg/ml extract of C.
aurantifolia inhibits the growth of colon SW-480
cancer cell in 78% aer 48 h of exposure. It increased
level of caspase-3. (72). ey also reported that C.
aurantifoliaextract can stop the growth of pancreatic
Panc-28 cancer cells with inhibitory concentration
50%, IC50, 18–42 µM. Among all the phytochemical
tested in study, the order of apoptosis was isolimonexic
acid > limonexic acid > sitosterol glucoside > limonin
> limonin glucoside, based on the expression ratio of
Bax/Bcl-2) (73).
Antioxidant property
Both fruit and peel juice of Citrus aurantifolia
posses numerous flavonoids that contribute to

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ARORA, P. et al - Braz. J. Nat. Sci. – electronic journal ISSN: 2595-0584 - V.3 - N.2
antioxidant effects of plant. Lime juice and peel
inhibits LDL oxidation in a dose dependent
manner. At 234 nm polyphenolic extract solution
of fresh lime juice (0-40 μl) prepared in DMSO,
showed significant antioxidant property measured
by lowry method (74). Patil and group revealed
that freeze-dried lime juice extracted with different
solvents, such as chloroform, acetone, methanol
and methanol/water (8:2). The chloroform extract
showed the highest (85.4 and 90%) radical-
scavenging activity analysed by 1,1-diphenyl-
2-picryl hydrazyl (DPPH) and 2,20 -azino-bis
(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS)
methods at 624 μg/ml (72). Limonoids possess the
ability to inhibit tumor formation by stimulating
the enzyme glutathione S-transferase (GST),
enzyme that catalyzes the reaction of glutathione
(75). Endogenously produced radical oxygen
species (ROS) perpetuate ongoing inflammation
that is a major factor in airway remodelling in
asthma. Vitamin C is a major antioxidant present
in airways. The plants enriched with phenolics
and ascorbic acid have shown promising results
in counteracting the radical production in lungs,
thereby, indicating prophylactic role of plant
in several diseases including, asthma (76-80).
Concentrated juice of C. aurantifolia cv. swingle
(Lime) at 250 µg/l, is able to significantly inhibit
proliferation of phytohaemagglutinin activated
mononuclear cells suggesting immunomodulating
activity of plant that suggests immuno-modulatory
property of plant (81, 82).
Anti-cholinesterase activity
e essential oils isolated from some species of C.
aurantifolia and C. aurantium have shown signicant
inhibitory activity on AChE and BChE with IC50
values of 139.3-147.5 μg/ml and 235.5 to 266.6 μg/ml
respectively (83, 84).
Anti-fertility activity
In experimental animals, oral administration
of undiluted lime juice to Sprague-Dawley female
rats has shown to compromise fertility by partially
blocking ovulation, ova formation (5.10 +/- 2.37)
in comparison with the control animals (12.70 +/-
1.14) (85).
Cardiovascular activity
C. aurantifolia is used in African folk medicine
for the management of hypertension. the eect was
validated in ex vivo studies conducted on isolated
heart of rabbit. Aqueous extract of C. aurantifolia
(10-80 mg/ml-10-20 mg/ml) produced both negative
inotropic and chronotropic eects on the heart
induced a dose-dependent relaxation of contractions
produced by adrenalin (3.10-3 mM) and KCl (80 mM)
(86). In cadmium induced hypertensive model of
spargue daweley rats, C. aurantifolia fruit extract, 0.75
g/kg, was able to successfully reduce both diastolic
and systolic blood pressure (87).
Conclusion
C. aurantifolia is valued for its nutritional qualities
and numerous health benets. e innumerable
health benets of C. aurantifolia and its essential oil
are attributed to multitude of bioactive compounds
including terpenes and phenolic components. is
opens new horizons for development of essential
oils into pharmaceutical products. However, lack
of scientic evidence to conrm medicinal value
warrants need of huge research in this direction.
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