International Journal of Bioassays : IJBNHY : 2278-778X

Abstract

Essential oils have found vast applications in many fields including aromatherapy, flavor and fragrance industries. Ixora coccinea Linn. is a reputable medicinal plant with long history of use in Nigeria. This study aimed to investigate the chemical constituents and antimicrobial activity of the leaf Essential Oil (EO) of Ixora coccinea grown in Nigeria. EO was obtained by hydrodistillation with yield of 0.16% (w/w). Chemical constituents of EO were determined using Gas Chromatography coupled to Mass Spectrometry (GC-MS). The GC-MS analysis identified 43 compounds, representing 94.67% of the oil constituents. The analysis revealed eight classes of compounds including hydrocarbons, alcohols, carboxylic acids, esters, aldehydes, ketones, sesquiterpenoids and triterpenoids. Hydrocarbons accounted for 33.77% with decane (11.12%) as the highest; alcohols comprised 28.86% of the oil with the highest being linalool (10.54%). Esters made up 14.15% of the oil. Carboxylic acid (10.91%) was dominated by malonic acid (10.26%); sesquiterpenoids made up 6.84% of the oil dominated by 3,7,11-trimethyl-1,6,10-dodecatrien-3-ol (3.07%). Aldehydes made up 3.36% of EO dominated by heptadecanal (2.30%). Ketones accounted for 1.38% of the oil. The inhibitory effect of EO was evaluated against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans and Mycobacterium tuberculosis (BCG) using broth microdilution method. The essential oil showed significant antimicrobial effects against the test organisms, with Minimum Inhibitory Concentration (MIC) ranging from 50 to 200 µg/mL. This result showed that the EO could serve as adjunct therapy in the treatment of community acquired infections.

Full text

International Journal of Bioassays
: IJBNHY
: 2278-778X
Open Access
*Corresponding Author:
Dr. Samuel Ehiabhi Okhale,
Department of Medicinal Plant Research and
Traditional Medicine, National Institute for
Pharmaceutical Research and Development, Idu
Industrial Area, P.M.B. 21 Garki, Abuja, Nigeria.
E-mail: samuelokhale@gmail.com
DOI: http://dx.doi.org/10.21746/ijbio.2018.7.5.1 pg. 5630
Research Article
1Department of Medicinal Plant Research and Traditional Medicine, National Institute for Pharmaceutical Research
and Development, Garki, Abuja, Nigeria
2Department of Microbiology and Biotechnology, National Institute for Pharmaceutical Research and Development,
Garki, Abuja, Nigeria
Received: 8/16/2018; Revised: 8/30/2018; Accepted: 9/07/2018
Available online: 12th September 2018
Abstract: Essential oils have found vast applications in many elds including aromatherapy, avor and fragrance
industries. Ixora coccinea Linn. is a reputable medicinal plant with long history of use in Nigeria. This study aimed to
investigate the chemical constituents and antimicrobial activity of the leaf Essential Oil (EO) of Ixora coccinea grown
in Nigeria. EO was obtained by hydrodistillation with yield of 0.16% (w/w). Chemical constituents of EO were
determined using Gas Chromatography coupled to Mass Spectrometry (GC-MS). The GC-MS analysis identied 43
compounds, representing 94.67% of the oil constituents. The analysis revealed eight classes of compounds including
hydrocarbons, alcohols, carboxylic acids, esters, aldehydes, ketones, sesquiterpenoids and triterpenoids. Hydrocarbons
accounted for 33.77% with decane (11.12%) as the highest; alcohols comprised 28.86% of the oil with the highest being
linalool (10.54%). Esters made up 14.15% of the oil. Carboxylic acid (10.91%) was dominated by malonic acid (10.26%);
sesquiterpenoids made up 6.84% of the oil dominated by 3,7,11-trimethyl-1,6,10-dodecatrien-3-ol (3.07%). Aldehydes
made up 3.36% of EO dominated by heptadecanal (2.30%). Ketones accounted for 1.38% of the oil. The inhibitory
effect of EO was evaluated against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans and
Mycobacterium tuberculosis (BCG) using broth microdilution method. The essential oil showed signicant antimicrobial
effects against the test organisms, with Minimum Inhibitory Concentration (MIC) ranging from 50 to 200 µg/mL. This
result showed that EO could serve as adjunct therapy in the treatment of community acquired infections.
Keywords: Ixora coccinea, Leaf, Essential oil, Antimicrobial
Introduction
Some medical and aromatic plants produce
fragrances and essence that are of immense benet
to man. Humans have exploited several plants
for these allures and essence termed “essential oil”.
Essential oils are volatile constituents extracted from
various parts of aromatic plants using mechanical
expression or hydrodistillation. Essential oils are
made up of a large array of chemical constituents
that consist essentially of terpenoids and non-
terpenoids found in various plant parts [1]. Herbal
therapy has gained popularity among physicians and
patients [2] as several medicinal and aromatic herbs
are loaded with metabolites that have demonstrated
several therapeutic effects [3].
Ixora, a genus of owering plants in the Rubiaceae
family consists of tropical evergreen trees that are
native to the tropical regions of Asia [4] comprising
about 500 different species with its centre of
diversity in Tropical Asia [5]. The word “Ixora” was
coined from a Portuguese version of Iswari, which
is the name of the Goddess “Parvati” to which Ixora
coccinea owers are offered, while “coccinea” is a
Latin word that means scarlet coloured [6]. Ixora
also grows in subtropical climates in the United
States, such as Florida and is distributed widely in
Nigeria [7,8]. The common names of Ixora coccinea
are Rangan, West Indian Jasmine, Kheme Chann
tanea, Jarum-jarum, Techi, Pan, Santan, Jungle ame
[7] and Jungle of Geranium or vetchi in Ayurveda
[4]. The plant Ixora coccinea (synonyms included
Ixora grandiora Bot and Ixora bandhuca Roxbg)
is cultivated as an ornamental plant. It is a multi-
branched, glabrous ever green shrub commonly 1-2
m in height, but capable of reaching up to 3.6 m
in height; bearing numerous bright scarlet coloured
owers which are in dense, senssile corymbiform
cymes. Leaves are coriaceous, from 2 cm to 15
cm in length, oblong, sessile or sub-sessile and
obtuse [9]. The plant is distributed preferably in
acidic soils and thrives in moist, well drained acidic
International Journal of Bioassays 7.5 (2018) pp. 5630-5637
Chemical constituents and antimicrobial activity of the leaf essential
oil of
Ixora coccinea L
(Rubiaceae) collected from North Central
Nigeria
Okhale SE1*, Ugbabe GE1, Oladosu PO2, Ib-rahim JA1, Egharevba HO1, Kunle OF1, Elisha EP1,
Chibuike AJ1, Ettah UO1,

Samuel EO International Journal of Bioassays 7.4 (2018) pp. 5630-5637
DOI: http://dx.doi.org/10.21746/ijbio.2018.7.1.1 pg. 5631
soil with tolerance for shade. Its pharmacological
prole include traditional uses as anti-mitotic,
hepatoprotective, antimicrobial, chemoprotective,
anti-oxidant, anti-nociceptive and anti-inammatory
activities [10]. The roots of I. coccinea is used as
antiseptic, astringent, stomachic, sedative and also
in the management of dysentery, diarrhea and
gonorrhea; anorexia, hiccups, sores, cough, fever
and chronic ulcers [11,12]. In Indo–china, root
decoction is useful in clarifying the urine and also
as poultice; fresh stems and leaves are used for
eczema, sprains, boils and contusions [6]. Flowers
are used in managing leucorrhoea, dysmenorrhea,
haemoptysis, dysentery and catarrhal bronchitis
[6,13,14]. The leaves have shown antimicrobial
[15-17], antinociceptive and anti-inammatory
properties [15,18]. The plant also has antioxidant
properties, anthelmintic activities, antileishmanial
activity, anti-asthmatic activity, anti-diarrhoeal
activity, hypoglycaemic and hypolipidaemic activity,
hepatoprotective activity, wound healing activity,
cytotoxic and antitumour activity, cardioprotective
activity, anti-ulcer activity, neuroprotective activity
and anxiolytic activity [15,17-35].
The leaves of Ixora coccinea yielded kaemferol,
avonoids, quercetin, anthrocyanidins, ferulic acids
and other phenolic acids [26]. The ower extract
contain avonoids, triterpenoids and tannins. The
owers are used topically to treat scabies, sores,
chronic ulcers and some type of dermatitis [16] and
also traditionally to enhance sexuality and the re-
kindling of passion [7]. Phytochemical investigation
of I. coccinea revealed important phytochemicals such
as ursolic acid, lupeol, oleanolic acid, rutin, sitosterol,
lecocyanadin, anthocyanins, proanthocyanidins,
quercetin, and kaempferol glycosides [7] many of
which have antimicrobial activities [17].
Phytochemical investigation of the root resulted
in the isolation of six phytoconstituents namely
β-amyrin, 9,12-octadecadienoic acid, kaempferitrin,
kaempferol-7-O-glucoside and quercetin [6,36]. The
root of I. coccinea also contained palmitic acid, stearic
acid, oleic acid, linoleic acid and mannitol [18,37].
Fifty-four components had been identied in
the essential oil of I. coccinea ower, representing
99.97% of the total components detected [7]. The
oil is composed mainly of triterpenes 62.60%,
monoterpenes 31.73%, sesquiterpenes 3.35% and an
ester 2.29%. The major triterpenes are ursolic acid
(27.34%), oleanolic (20.16%) and lupeol (15.10%).
Geranyl acetate (8.74%) is the major monoterpenes,
followed by linalyl acetate (6.79%), neryl acetate
(6.49%), terpineol acetate (4.91%), and borneol
acetate (4.77%); ethyl cinnamate (2.29%) an ester
while the sesquiterpenes are cyperene (2.72%) and
α–copaene (0.63%) [38]. A new triterpene, ixorene
with dammarane skeleton has been isolated from the
leaves of I. coccinea, along with β-sitosterol, lupeol
and D-mannitol [7]. Furthermore, the air-dried
owers of I. coccinea afforded cycloartenol esters,
lupeol fatty ester, lupeol, ursolic acid, oleanolic acid
and sitosterol [39].
A new natural terpenoid, ixoroid, was isolated from
the ower of Ixora coccinea along with the known
constituents stigmast-5-en-3-O-β-D-glucoside, 5-O-
caffeoylquinic acid and D-mannitol [40]. In addition,
ixorapeptide I and ixorapeptide II [41], as well as
compounds like biochin A, myricetin, quercetin, rutin,
diadzein and formononetin from the methanolic
ower extract had been reported [42]. The leaves of
Ixora coccinea had been reported to contain Ixora tannin
A-2 (a trimeric A-type proanthocyanidin), procyanidin
A2, cinnamtannin B-1 [43,44].
The increasing prevalence of multi-drug drugs
resistant strains of bacteria and the recent
appearance of strains with reduced susceptibility to
clinically used antibiotics has led to the quest for new
antimicrobial agent from plant sources to overcome
this challenge. Ixora coccinea L is a reputable medicinal
plant in North Central Nigeria, where it is used for
the treatment of infectious diseases, among other
ailments. There is no reported work on the chemical
constituents and antimicrobial activity of the leaf
essential oil of Ixora coccinea from North Central
Nigeria. The aimed of this study was to investigate
the chemical composition and antimicrobial activity
of the leaf essential oil of Ixora coccinea collected
from North Central Nigeria.
Materials and Methods
Plant materials and volatile oil Collection
The plant Ixora coccinea was collected in July at the
NIPRD Garden, Abuja, Nigeria. The plant was
identied by a taxonomist at the herbarium of
the National Institute of Pharmaceutical Research
Development, Abuja, Nigeria, where a voucher
specimen (NIPRD/H/5229) was deposited.
Fresh leaves of Ixora coccinea (500 g) were chopped
into small pieces and the material was then
hydrodistilled for 4 h using Clavenger type apparatus.
Light-yellow coloured oil in yield of 0.16% (w/w)
was obtained. The volatile oil obtained was dried
over anhydrous sodium sulphate and stored at 4ºC
in sealed vials until analysis.
The essential oil was analyzed by GC-MS using
Gas Chromatography–Mass Spectrometry (GC-
MS) analyses

Samuel EO International Journal of Bioassays 7.4 (2018) pp. 5630-5637
DOI: http://dx.doi.org/10.21746/ijbio.2018.7.1.1 pg. 5632
SN Name of compound Classication RT (min) % Composition
1 Cyclohexanepropanol Alcohol 3.286 4.67
2 2-methylpropyl cyclohexane Cyclic hydrocarbon 3.495 1.20
3 Malonic acid, 2-heptyl tetradecyl ester Carboxylic acid 3.722 10.26
4 Mesitylene Aromatic hydrocarbon 4.116 6.13
5 Decane Saturated hydrocarbon 4.192 11.12
6 1,3,3-trimethylnonyl benzene Aromatic hydrocarbon 4.493 3.16
7 Linalool Alcohol 5.495 10.54
8 Methyl salicylate Ester 6.827 3.40
9 Geraniol Terpene alcohol 7.617 2.42
10 Ionone Ketone 8.045 0.02
11 Citronellyl acetate Ester 8.931 0.52
12 β-Damascenone Ketone 9.411 0.41
13 Tetradecane Saturated hydrocarbon 9.603 1.41
14 5,9-Undecadien-2-one,6,10-dimethyl- Ketone 10.218 0.95
15 1-Dodecanol Alcohol 10.503 1.67
16 β-Selinene Aromatic hydrocarbon 10.837 2.30
17 α-Selinene Sesquiterpenoid 10.945 1.60
18 (-)-Spathulenol Sesquiterpenoid alcohol 11.320 0.63
19 Nerolidol Sesquiterpene 11.631 3.07
20 cis-3-Hexenyl benzoate Ester 11.709 1.89
21 Supraene Triterpenoid 11.807 0.94
22 Hexadecane Saturated hydrocarbon 12.055 1.42
23 Tetradecanal Aldehyde 12.113 0.12
24 1-Hexadecanol Alcohol 13.004 1.50
25 Heptadecanal Aldehyde 13.554 2.30
26 Hexadecane Saturated hydrocarbon 14.937 0.79
27 Isopropyl myristate Ester 15.233 0.59
28 6,10,14-Trimethyl-2-pentadecanone Sesquiterpenoid 15.488 2.17
29 Benzyl salicylate Ester 15.801 1.11
30 1-Hexadecanol Alcohol 15.976 0.29
31 cis,cis,cis-7,10,13-Hexadecatrienal Aldehyde 16.041 0.94
32 Hexadecanoic acid, methyl ester Ester 16.312 0.41
33 n-Hexadecanoic acid Carboxylic acid 16.715 0.65
34 n-Nonadecanol Alcohol 17.070 4.84
35 2-Methylhexacosane Acyclic hydrocarbon 17.134 0.84
36 Isopropyl palmitate Ester 17.324 1.12
37 Geranylgeranylacetate Ester 17.479 0.13
38 Methyl-6-octadecenoate Ester 17.908 0.68
39 Phytol Alcohol 18.036 0.31
40 Nonadecene Unsaturated hydrocarbon 18.666 1.46
41 Tetracosane Saturated hydrocarbon 19.324 0.95
42 1,54-Dibromotetrapentaconntane Hydrocarbon 21.748 2.78
43 2,2,4-Trimethyl-3-(3,8,12,16-tetramethyl-heptadeca-
3,7,11,15-tetraenyl]-cyclohexanol Alcohol 22.751 1.99
Total 94.67%
RT=Retention time.
Shimadzu QP-2010 GC with QP-2010 Mass
Selective Detector [MSD, operated in the EI mode
(electron energy=70 eV), scan range of 45-400 amu,
and scan rate of 3.99 scans/sec], and Shimadzu GC-
MS solution data system. The Gas chromatography
column was Optima-5 ms fused silica capillary with
5% phenyl-methylpolysiloxane stationary phase, with
length of 30 m, internal diameter of 0.25 mm and
lm thickness of 0.25 μm. The carrier gas was helium
with ow rate of 1.61 mL/min. The program used
for Gas chromatography oven temperature was 60-
180°C at a rate of 10°C/min, then held at 180°C for
2 min, followed by 18-280°C at a rate of 15°C/min,
then again held at 280°C for 4 min. The injection port
temperature was 250°C while detector temperature
was 280°C. Helium was used as a carrier gas, at a ow
rate 1.61 mL/min. Diluted sample (1/100 in hexane,
v/v) of 1.0 μL was injected using autosampler and
in the split mode with ratio of 10:90. Individual
constituents were identied by comparing their mass
spectra with known compounds and NIST Mass
Spectral Library (NIST 11). The percentages of each
component are reported as raw percentages based
on the total ion current without standardization. The
essential oil constituents of Ixora coccinea leaf is as
detailed in Table 1.
Table 1: Percentage of compounds in the volatile oil of Ixora coccinea leaf.

Samuel EO International Journal of Bioassays 7.4 (2018) pp. 5630-5637
DOI: http://dx.doi.org/10.21746/ijbio.2018.7.1.1 pg. 5633
Microbial strains
The following microorganisms were used in the
evaluation of the antibacterial activity of the
essential oil: Gram-positive bacteria Staphylococcus
aureus (ATCC 25923); Gram-negative bacteria,
Pseudomonas aeruginosa (ATCC 27853), Klebsiella
pneumonia (ATCC 13883), Escherichia coli (ATCC
10798), and fungi Candida albicans (ATCC 2876).
Antimicrobial activity
The Minimum Inhibitory Concentration (MIC)
values of the essential oil of Ixora coccinea leaf were
determined in triplicate by the broth microdilution
method in 96-well microplates. The oil sample was
dissolved in Dimethyl Sulfoxide (DMSO) followed
by addition of sterile Mueller-Hinton broth for
bacteria and Saboraud-Dextrose broth for Candida
albicans, to achieve concentration of 200 μg/mL.
The nal DMSO concentration was 10% (v/v)
and this solution was used as a negative control.
The inoculum was adjusted for each organism
to yield a cell concentration of 2 × 10-7 colony
forming units (cfu) per mL. Ciprooxacin (Fidson,
Lagos Nigeria) was used as a positive control for
bacteria and Fluconazole (Pzer, UK) was used as
the standard drug for fungi at stock concentration
of 50 μg/mL. Controls of sterility for the Mueller-
Hinton nutrient broth, control culture (inoculum),
ciprooxacin, uconazole, essential oil and DMSO
were performed. The microplates were closed and
incubated aerobically at 37°C for 24 h. The MIC
values were determined as the lowest concentration
of essential oil capable of inhibiting the growth of
the microorganisms. All assays were carried out in
triplicate. Results are shown in Table 2.
Results and Discussion
Plants contain a vast array of secondary
metabolites with diverse pharmacological activities
[45,46]. Among these secondary metabolites are
essential oils. The application of essential oils in
aromatherapy for treating different ailments has
been reported [47-49]. The antimicrobial activities
of essential oils from plants had been reported [50 -
53] and Ixora coccinea leaf extracts had been reported
to have antimicrobial properties [54]. This study
focused on evaluating the chemical constituents
and antimicrobial activity of the leaf essential oil of
Ixora coccinea grown in Nigeria.
The essential oil isolated from the leaves have
characteristic odour and a light yellow colour.
Further analysis using GC-MS resulted in the
identication of 43 compounds as shown in (Table
1) representing 94.67% of the total essential oil
constituents. Essential oil formation is affected
by seasonal variation, climatic conditions such as
temperature, sunlight, frequency and magnitude
of precipitation and time of harvesting. The leaf
volatile oil composed of several classes of chemicals
such as alcohols (also consisting monoterpene and
sesquiterpene alcohols) hydrocarbons; carboxylic
acids, esters, aldehydes, ketones, sesquiterpenoids
and triterpenoids. The leaf oil was highest in
hydrocarbons (cyclic, aromatic, saturated and
unsaturated hydrocarbons), amounting to 33.77%
of the total constituents such as decane (11.12%),
mesitylene (6.13%), cyclohexane (1.20%), benzene
(3.16%), tetradecane (1.41%), naphthalene (2.30%);
hexadecane (2.21%), 2-methylhexacosane (0.84%),
nonadecane (1.46%), tetracosane (0.95%) and
tetrapentacontane,1,54-dibromo (2.99%)). It has
been reported that steam-distillation and hydro-
distillation methods yield oils rich in terpene
hydrocarbons, while in contrast, the super-critical
extracted oils contained a higher percentage of
oxygenated compounds [55–58] which implies
that the qualitative and quantitative chemical
composition of the essential oil differs according to
the technique of extraction applied.
The second largest total constituent was alcohol
(28.86%) comprising of linalool (10.54%),
cyclohexanepropanol (4.67%), geraniol (0.74%),
geraniol, a terpene alcohol (2.42%), dodecanol
(1.67%), sputhulenol (0.63%), hexadecanol (1.79%),
n-nonadecanol (4.84%), phytol (0.31%) and
2,2,4-trimethyl-3-(3,8,12,16-tetramethyl-heptadeca-
3,7,11,15-tetraenyl]-cyclohexanol (1.99%). The
leaf oil had esters of fatty acid and carboxylic acid
S/N Micro-organisms Minimum inhibitory concentrations (MIC values μg/mL)
Essential oil Standard drug
1* Pseudomonas aeruginosa (ATCC 27853) 100 0.05
2* Klebsiella pneumonia (ATCC 13883) 50 0.39
3* Escherichia coli (ATCC 10798) 100 0.39
4* Staphylococcus aureus (ATCC 25923) 200 0.10
5** Candida albicans (ATCC No. 2876) 100 6.25
7*** Mycobacterium bovis BCG (ATCC 27290) 200 0.04
*Bacterial strain; **fungal strain; ***mycobacterium strain. Ciprooxacin (standard drug for bacterial strains); uconazole (standard drug for fungal strain);
isoniazid (standard drug for mycobacterium strain)
Table 2: Inhibitory effects on the growth of bacteria (MIC values μg/mL) of the volatile oil from the leaf of Ixora coccinea.

Samuel EO International Journal of Bioassays 7.4 (2018) pp. 5630-5637
DOI: http://dx.doi.org/10.21746/ijbio.2018.7.1.1 pg. 5634
which made up 14.15% of the total components
of the volatile oil such as methylsalicylate (3.40%),
citronellyl acetate (0.52%), 3-hexenyl benzoate
(1.89%), isopropl myristate (0.59%), benzyl salicylate
(1.11%), methyl hexadecanoate (0.41%), isopropyl
palmitate (1.12%); geranylgeranylacetate (0.13%),
methyl-6-octadecenoate (0.68%). Aldehydes made
up 3.36% of the total oil constituents and included
tetradecanal (0.125%), heptadecanal (2.30%) and
7,10,13-hexadecatrienal (0.94%). The percentage
content of sesquiterpenoids was 6.84% of the total
volatile oil components and comprised of α-selinene
(1.60%); 3,7,11-trimethyl-1,6,10-dodecatrien-3-ol
(3.07%) and 6,10,14-trimethyl-2-pentadecanone
(2.17%). Ketones accounted for 1.38% of the leaf
volatile oil constituents and comprised ionone
(0.02%); 6,10-dimethyl-5,9-undecadien-2-one
(0.95%) and β-damascenone. Triterpenoids made
up 0.94% of the total essential oil constituents and
comprised of supraene (0.94%); carboxylic acids
were malonic acid (10.26%) and hexadecanoic acid
(0.65%).
Sesquiterpenes are known to delete bad information
from the cell’s memory and is useful as xative in
perfume industry while monoterpenes inhibits
the accumulation of toxin in the body [59].
Methylsalicylate is used as fragrance in beverages
and food products [60,61] and also used as an
antimicrobial agent [61]. The antibacterial activity of
some monoterpenes, diterpenoids, sesquiterpenes,
triterpenoids and their derivatives had been reported
[62]. Diterpenoids and sesquiterpenes isolated from
different plants inhibited the growth of Mycobacterium
tuberculosis [63, 64] and exhibited bactericidal activity
against Gram-positive bacteria. The mechanism
of action of terpenoids is not fully understood,
but speculated to involve membrane disruption by
the lipophilic compounds. Sesquiterpenoids have
been reported to have antimicrobial properties
[65]; and alcohols such as phytol have antibacterial
activity against Pseudomonas aeruginosa [66]; geraniol,
occurring in the essential oils of several aromatic
plants is most important molecules in the avour
and fragrance industries because of its pleasant
odour, geraniol is also known to have antimicrobial
activities [67]. The preserving quality of benzoic
acid is due to its ability to delay the multiplication
of several groups of microorganisms making its
action bacteriostatic [68]. Most of the antimicrobial
activities of essential oils were attributable to the
oxygenated terpenoids, while some hydrocarbons
also exhibited antimicrobial effects [69–71].
Interactions between the different components may
lead to additive, antagonistic or synergistic effects.
The susceptibility of some micro-organisms were
evaluated against the essential oil obtained from the
leaves of Ixora coccinea (Table 2). The MIC values
were determined as the lowest concentration of
the essential oil capable of inhibiting the microbial
growth. Klebsiella pneumonia had Minimum Inhibitory
Concentration (MIC) of 50 μg/mL); Pseudomonas
aeruginosa with MIC of 100 μg/mL; Escherichia coli
with MIC of 100 μg/mL; Staphylococcus aureus with
MIC of 200 μg/mL; Candida albicans with MIC of
100 μg/mL and Mycobacterium tuberculosis (BCG)
with MIC of 200 μg/mL. The test pathogens are
contaminants in food, man as well as animals and
air [72]. The essential oil of Ixora coccinea was more
active against Klebsiella pneumonia with MIC of 50
μg/mL. The antibacterial activity exhibited by Ixora
coccinea leaf essential oil supports its use in traditional
medicine for treatment of infectious diseases.
Conclusion
Essential oils are natural plant products containing
complex mixture of components and thus
have multiple antimicrobial properties with the
interactions between these components resulting
in antagonism, additive or synergistic effects as
observed in the leaf essential oil of Ixora coccinea.
The antibacterial activity exhibited by the essential
oil suggested that it can be exploited in herbal
medicine for the management of respiratory and
gastrointestinal diseases.
References:
1. U.S. National Library of Medicine:Aromatherapy
and Essential Oils (PDQ®). PDQ Integrative,
Alternative, and Complementary Therapies
Editorial Board. 2018.
2. Jurga B, Ruta M, Julija D, Rimantas P, Romualda
G, et al. Topical application of Calendula
ofcinalis (L): Formulation and evaluation of
hydrophilic cream with antioxidant activity. J
Med Plant Res. 5.6 (2011): 868-877.
3. Chih-chien L, Chao-Hsun Y, Pey-Shiuan W,
Chang-Chin K, Yi-Shyan C. Antimicrobial,
antityrosinase and antioxidant activities of
aqueous aromatic extracts from forty- eight
selected herbs, J Med Plant Res. 5.26 (2011):
6203-6209.
4. Neelamegam R . Allelopathic effect of Ixora
coccinea Linn. on seed germination and early
seedling growth of paddy (Oryza sativa L.). J
Phytol. 3.6 (2011): 51-55.
5. Varier VPS. Indian Medicinal Plants, a
compendium of 500 species, University press
Pvt. Ltd, Hyderabad. (2010): 239.

Samuel EO International Journal of Bioassays 7.4 (2018) pp. 5630-5637
DOI: http://dx.doi.org/10.21746/ijbio.2018.7.1.1 pg. 5635
6. Joshi AB, Surlikar PM, Bhobe M. Ixora coccinea
Linn: Phytochemical Investigation. Int J Res
Pharm Chem. 3.3 (2013): 691-696.
7. Sunitha D, Hemalatha K, Bhagavanraju M.
Phytochemical And Pharmacological Prole of
Ixora: a review. Int J Pharm Sci Res. 6.2 (2015):
567-584
8. Obuzor GU, Nwakanma GU. Chemical
composition of essential oil of Ixora coccinea
ower from Port Harcourt, Nigeria. Int J Aca
Res. 3 (2011): 381–384.
9. Kharat AR, Nambiar VV, Tarkasband YS,
Pujari RR. A Review on Phytochemical and
Pharmacological activity of genus Ixora. Int J
Res Pharm Chem. 3.3 (2013): 628-635.
10. Elumalai A, Chinna E, Yetcharla V, Burle Shiva
K, Chava N. Phytochemical and pharmacological
prole of Ixora coccinea Linn. Int J Pharm Life
Sci. 3.3 (2012):1563-1567.
11. Nayak BS, Udupa AL, Udupa SL. Effect of
Ixora coccinea owers on dead space wound
healing in rats. Fitoterapia. 70 (1999): 233–236.
12. Aranachalam G, Subramanian N, Pazhani GP,
Karunanithi M, Ravichandran V. Phytochemical
and anti-ulcer investigations of the fresh leaf
extract of Ixora coccinea Linn (Rubiaceae) in
albino rat model. Int. J Pharma Sci.1 (2009): 26-
31.
13. Banerjee S, Chanda A, Ghoshal A, Debnath S,
Das A, et al. Nitric oxide scavenging activity
study of ethanolic extracts of Ixora coccinea
from two different areas of Kolkata. Asian J
Exp Biol Sci. 2.4 (2011): 595-599.
14. Khare CP. Indian Medicinal Plants. Springer
(India) Pvt.Ltd. New Delhi. 2007: 338-339.
15. Nripendra NB, Deb KS, Sudipta C, Partha PM
. Comparative study of in vitro free radical
scavenging activity of different leaf extracts of
Ixora coccinea L. Int J Biomed Res. 2.1 (2011):
32-40.
16. Mukesh CS, Smita S. Preliminary Phytochemical
and antimicrobial investigations of the aqueous
extract of Ixora coccinea L. and Commelina
benghalensis L. on Gram-Positive and Gram-
Negative Microorganisms. Middle East J Sci Res.
6.5 (2010): 436-439.
17. Nagaraj B, Krishnamurthy N, Liny P, Divya T,
Dinesh R. Biosynthesis of Gold Nanoparticles
of Ixora coccinea ower extract & their
Antimicrobial activities. Int J Pharm Bio Sci. 2.4
(2011): 557-565.
18. Baliga MS, Kurian PJ. Ixora coccinea L., traditional
uses, phytochemistry and pharmacology. Chin J
Integr Med. 18.1 (2012): 72-79.
19. Rohini S, Shalini N, Narayanaswamy,
Balakrishnan KP. Application of Natural
Products In Cosmetics: A Study of Ixora coccinea
Extracts for their Antityrosinase and Antioxidant
Activities. Int J Res Cos Sci. 2.1 (2012): 1-7.
20. Sankhadip B, Sushomasri M, Pranabesh C.
Comparative study of in vitro and in vivo
antioxidant property of different Ixora species. J
Adv Pharm Edu & Res. 2 (2011): 90-103.
21. Surana AR, Aher AN, Pal SC, Deore. Evaluation
of anthelmintic activity of Ixora coccinea. J Pharm
Life Sci. 2011; 2(6): 813-814.
22. Naskar M, Bhattacharya S, Biswas M.
Antileishmanial effect of Ixora coccinea leaf
extracts on the in vitro growth of Leishmania
donovani promastigotes. J Adv Pharm Edu Res.
3.4 (2013): 471-474.
23. Missebukpo A, Metwogo K, Agobon A, Eklu
Gadegbeku K, Akilikoku K, et al. Evaluation of
anti-asthmatic activity of Ixora coccinea. J Pharma
Toxicol. 6.6 (2011): 559-570.
24. Maniyar Y, Bhixavatimath P, Agashikar NV.
Anti-diarrhoeal activity of owers of Ixora
coccinea Linn. in rats. J Ayurveda Integr Med.
1.4 (2011): 287-291.
25. Prabhu B, Yasmeen M, Agashikar NV.
Evaluation of the Anti-diarrhoeal Activity of
the Leaves of Ixora coccinea L. in rats. J Clin
Diagno Res. 4 (2010): 3298-3303.
26. Yasmeen M, Prabu B. Evaluation of the
Hypoglycaemic and Hypolipidaemic activities
of the aqueous extract of the leaves of I. coccinea
L. in Diabetic Rats. J Clin Diagno Res. 5.7
(2011): 1381-1384
27. Shyamal S, Latha PG, Suja SR, Shine VJ, Anuja
GI, et al. Hepatoprotective effect of three
herbal extracts on aatoxin B1- intoxicated rat
liver. Singapore Med J. 51.4 (2010): 326-331.
28. Selvaraj N, Lakshmanan B, Mazumder PM,
Karuppasamy M, Jena SS, et al. Evaluation of
wound healing and antimicrobial potentials of
Ixora coccinea root extract. Asian Pac J Trop
Med. 4.12 (2011): 959-963.
29. Aadesh U, Pronobesh C, Danswrang G, Papiya
MM, Vijay V. Ixora coccinea enhances cutaneous
wound healing by upregulating the expression
of collagen and basic broblast growth factor.
ISRN Pharmacology. (2014): 1-9.

Samuel EO International Journal of Bioassays 7.4 (2018) pp. 5630-5637
DOI: http://dx.doi.org/10.21746/ijbio.2018.7.1.1 pg. 5636
30. Saravanan P, Boopalan E. Occurrence of
Camptothecin an Anticancer Drug from Ixora
coccinea Linn. Int J App Biol. 2.2 (2011): 30-34.
31. Firoz NM, Bharatesh RK, Tabassum SS, Nilofar
SN. Cardioprotective effect of methanolic
extract of Ixora coccinea L. leaves on doxorubicin-
induced cardiac toxicity in rats. Indian J
Pharmacol. 44.2 (2012): 178–183.
32. Arunachalam G, Subramanian N, Pazhani P,
Karunanithi M, Ravichandran V. Phytochemical
and anti-ulcer investigations of the fresh leaf
extract of Ixora coccinea L (Rubiaceae) in albino
rat model. Int J Pharm Sc.1.1 (2009): 26-31.
33. Patibandla NB, Nagaraju B, Vinay I. Evaluation
of antiulcer and in-vitro antioxidant activities of
Ixora coccinea owers and polyherbal extract in
Wistar Albino Rats. Int J Pharm Pharm Sci. 6.2
(2014): 339-344.
34. Harish KS, Srinath R. Evaluation of
Neuroprotective Activity of Ixora coccinea extract
on experimentally induced Neurotoxicity in
Albino Rats. Synopsis 2013. Rajiv Gandhi
University of Health Sciences, Bengaluru,
Karnataka.
35. Shan PM, Nasiya L, Sri GP. Evaluation of
Anxiolytic activity of Ixora coccinea L. ethanolic
extract in Swiss Albino mice. Clin Exp
Pharmacol. 4.146 (2014): 1-3.
36. Kartha ARS, Menon KN. The isolation and
constitution of an acid from the root bark of
Ixora coccinea L. Proceedings of Mathematical
Sciences. (1943): 11–15.
37. Yadava RN. Analysis of Fixed Oils from the
roots of I. coccinea. Asian J Chem. 1 (1989): 307-
308.
38. Gloria UO, Gibson UN. Chemical composition
of Essential oil of Ixora coccinea ower from Port
Harcourt, Nigeria. Int J Acad Res. 2. (2011): 381.
39. Ragasaa CY, Tiua F, Rideoutb JA. New
Cycloartenol esters from I. coccinea. Nat Prod
Res. 18.4 (2004): 319–323.
40. Versiania MA, Ikrama A, Khalida S, Faizib S,
Tahiria IA. Ixoroid: A New Triterpenoid from
the Flowers of Ixora coccinea. Nat Prod Commun.
7.7 (2012): 831-834.
41. Lee C, Liao Y, Hwang T, Wu C, Fang-Rong et
al. Ixorapeptide I and Ixorapeptide II, bioactive
peptides isolated from Ixora coccinea. Bioorganic
& Medicinal Chemistry Letters. 20 (2010): 7354–
7357.
42. Sumathy H, Sangeetha J, Vijayalakshmi K.
Chromatographic Fingerprint Analysis of Ixora
coccinea Methanolic Flower Extract. Int J Pharma
Sci Drug Res. 3.4 (2011): 327-330.
43. Ambreen I, Muhammad AV, Shumaila S, Salman
KA, Syed TA, et al. Ixorene, a new Dammarane
triterpene from the leaves of Ixora coccinea L. Rec
Nat Prod. 7.4 (2013): 302-306.
44. Idowu TO, Ogundaini AO, Salau AO, Obuotor
EM, Bezabih M, et al. Doubly linked, A-type
proanthocyanidin trimer and other constituents
of Ixora coccinea L. leaves and their antioxidant
and antibacterial properties. Phytochemistry. 71
(2010): 2092–2098.
45. Assob JC, Kamga HL, Nsagha DS. Antimicrobial
and toxicological activities of ve medicinal
plant species from Cameroon Traditional
Medicine. BMC Complement. Altern Med. 11.
(2011): 70-81.
46. Arruda AL, Vieira CJ, Sousa DG, Oliveira RF,
Castilho RO. Bignoniaceae as antibacterial agent.
J Med Food. 14 (2011): 1604-1608.
47. Silva-Néto RP, Peres MF, Valença MM. Odorant
substances that trigger headaches in migraine
patients. Cephalalgia. 34.1 (2014): 14-21.
48. Vethanayagam D, Vliagoftis H, Mah D, Jeremy
Beach, Ladd Smith, et al. Fragrance materials in
asthma: a pilot study using a surrogate aerosol
product. J Asthma. 50.9 (2013): 975-982.
49. Celeiro M, Guerra E, Lamas JP, Marta Lores,
Maria Llompart. Development of a multianalyte
method based on micro-matrix-solid-phase
dispersion for the analysis of fragrance allergens
and preservatives in personal care products. J
Chromatogr A. (2014): 1344: 1-14.
50. Radaelli M , Silva BP, Weidlich L, Hoehne L,
Luiz AF, et al. Antimicrobial activities of six
essential oils commonly used as condiments
in Brazil against Clostridium perfringens BMC
Complement. Altern Med. (2006): 6:39.
51. Prabuseenivasan S, Jayakumar M, Ignacimuthu
S. In vitro antibacterial activity of some plant
essential oils. Asian Pac J Trop Biomed. 9.2
(2012): 739–742.
52. Bachir RG, Benali M. Antibacterial activity
of the essential oils from the leaves of
Eucalyptus globulus against Escherichia coli
and Staphylococcus aureus. Asian Pac J Trop
Biomed. 9.2 (2012): 739-742.
53. Marcela R, Silva BP, Luciana Weidlich, Lucélia
Hoehne, Eduardo Miranda Ethur: Antimicrobial

Samuel EO International Journal of Bioassays 7.4 (2018) pp. 5630-5637
DOI: http://dx.doi.org/10.21746/ijbio.2018.7.1.1 pg. 5637
activities of six essential oils commonly used
as condiments in Brazil against Clostridium
perfringens. Brazilian Journal of Microbiology.
47.2 (2016): 424-430.
54. Annapurna J, Amarnath PV, Amar D,
Ramakrishna SV, Raghavan KV. Antimicrobial
activity of Ixora coccinea leaves. Fitoterapia. 74.3
(2003): 291-293.
55. Donelian A, Carlson LHC, Lopes TJ, Machado
RAF. Comparison of extraction of patchouli
(Pogostemon cablin) essential oil with supercritical
CO2 and by steam distillation. J Supercrit Fluids.
48 (2009): 15-20.
56. Eikani MH, Golmohammad F, Rowshanzamir
S. Subcritical water extraction of essential oils
from coriander seeds (Coriandrum sativum L.). J
Food Eng. 80 (2007): 735-740.
57. Reverchon E. Supercritical uid extraction
and fractionation of essential oils and related
products. J Supercrit Fluids. 10 (1997):1-37.
58. Wenqiang G, Shufen L, Ruixiang Y, Shaokun T,
Can Q. Comparison of essential oils of clove
buds extracted with supercritical carbon dioxide
and other three traditional extraction methods
chemistry of essential oils. Food Chem.101
(2007): 1558-1560.
59. Solo Build It-SBI. The chemistry of essential
oil reveals aromatherapy as a true science. 2010-
2017.
60. Mason L, Moore RA, Edward JE, McQuay HJ,
Derry S, et al. Systemic Review of efcacy of
topical rubefacients containing salicylates for
the treatment of acute and chronic pains. BMJ.
328.7446 (2004): 995.
61. Oloyede GK. Toxicity, antimicrobial and
antioxidant activities of methylsalicylate
dominated essential oils of Laportea aestuans
(Gaud). Arabian J Chem. 9.1 (2011): S840-S845.
62. Suryakala G, Archana G. Production and
quantication of Asiatic acid from in vitro raised
shoots and callus cultures of Centella asiatica (L.).
urban. Ann Phytomed. 2.1 (2013): 95-101.
63. Garcia A, Bocanegra GV, Palma-Nicolas JP,
Rivera G. Recent advances in antitubercular
natural products. Eur J Med Chem. 49 (2012):1-
23.
64. Kurek A, Grudniak AM, Kraczkiewicz DA,
Wolska KI. New antibacterial therapeutics and
strategies. Pol J Microbiol. 60 (2011): 3-12.
65. Fortuna AM, Juárez ZN, Bach H, Hernandez
JR. Antimicrobial activities of sesquiterpene
lactones and inositol derivatives from
Hymenoxys robusta. Phytochemistry. 72.18
(2011): 2413-2418.
66. Lee W, Woo E, Lee DG. Phytol has antibacterial
property by inducing oxidative stress response I
Pseudomonas aeruginosa. Free Radic Res. 50.12
(2016): 1309-1318.
67. Chen W, Viljoen AM. Geraniol - A review of
a commercially important fragrance material. S
Afr J Bot. 74.6 (2010): 643-651.
68. Jorge K. in Encyclopedia of Food Sciences and
Nutrition (Second Edition). 2003.
69. Delaquis PJ, Stanich K, Girard B, Mazza,
G. Antimicrobial activity of individual and
mixed fractions of dill, cilantro, coriander and
eucalyptus essential oils. Int J Food Micro. 74
(2002): 101–109.
70. Burt S. Essential oils: Their antimicrobial
properties and potential applications in foods:
A review. Int J Food Microbiol. 94.3 (2004):
223–253.
71. Koroch A, Juliani HR, Zygadlo JA. Bioactivity of
Essential Oils and Their Components. In Flavours
and Fragrances Chemistry. Bioprocessing and
Sustainability, Berger, R.G., Ed., Springer Verlag:
Berlin, Germany. (2007): 87–115.
72. Okhale SE, Buba CI, Oladosu P ,Okoro IJ,
Ugbabe GE, et al. Chemical constituents and
antibacterial activities of the volatile oils of
Crinum zeylanicum L. and Crinum ornatum
(AIT) Bury bulbs. Int J Mod Pharma Res. 7.2
(2018): 01-07.
Cite this article as:
Okhale SE, Ugbabe GE, Oladosu PO, Ibra-him JA,
Egharevba HO, et al. Chemical constituents and antimi-
crobial activity of the leaf essential oil of Ixora coccinea
L (Rubiaceae) collected from North Central Nigeria.
International Journal of Bioassays 7.5 (2018) pp. 5630-5637. DOI:
http://dx.doi.org/10.21746/ijbio.2018.7.1.1