Phytomedicine 9: 259–262, 2002

© Urban & Fischer Verlag

http://www.urbanfischer.de/journals/phytomed
Phytomedicine

Formulation of an effective mosquito-repellent topical

product from Lemongrass oil
A. O. Oyedele1, A. A. Gbolade2, M. B. Sosan3, F. B. Adewoyin4, O. L. Soyelu3,
and O. O. Orafidiya1
1
Department of Pharmaceutics, 2Department of Pharmacognosy, 3Department of Plant Science and the 4Drug Research and
Production Unit, Obafemi Awolowo University, Ile-Ife, Nigeria

Summary
Ointment and cream formulations of lemongrass oil in different classes of base and the oil in liquid
paraffin solution have been evaluated for mosquito repellency in a topical application. Mosquito
repellency was tested by determining the bite-deterrence of product samples applied on an experi-
mental bird’s skin against a 2-day starved culture of Aedes aegypti L. mosquitoes. The 1%v/v solu-
tion and 15%v/w cream and ointment preparations of the oil exhibited ≥50% repellency lasting
2–3 h, which may be attributed to citral, a major oil constituent. This activity was comparable to
that of a commercial mosquito repellent. Base properties of the lemongrass oil formulations influ-
enced their effectiveness. The oil demonstrated efficacy from the different bases in the order of hy-
drophilic base > emulsion base > oleaginous base.

Key words: Cymbopogon citratus, essential oil, mosquito repellency, formulation.

j Introduction
Lemongrass oil is the essential oil obtained from the
aerial parts of Cymbopogon citratus (DC) Stapf., fami-
ly Poaceae. The plant has been widely recognized for
its ethnobotanical and medicinal usefulness (Dalziel,
1937). The insecticidal (Arias et al., 1992), antimicro-
bial (Syed et al., 1995), and therapeutic properties (Ak-
endengue, 1992) of its oil and extracts have been re-
ported. Trado-medicinal preparations of the oil have
been used both internally for alleviating colds and
fever symptoms (Comerford, 1996) and externally to
treat skin eruptions, wound and bruises (Spring, 1989).
Plant essential oils in general have been recognized as
an important natural resource of pesticides – insecti-
cides (Raguraman and Singh, 1997; Gbolade, 2001),
larvicides (Adebayo et al., 1999), and repellents
(Thorsell et al., 1998; Oyedele et al., 2000). The repel-
lents are designed as topical preparations or com-
bustible products that are able to protect the user or en-
vironment from harmful insects, such as mosquitoes,
which transmit diseases through their bite. The G.C.
analysis of Indian Cymbopogon essential oil as well as
its repellent effects against mosquito vectors of malaria
(Anopheles stephensi), filariasis (Culex quinquefascia-
tus), and dengue fever (Aedes aegypti) have been de-
scribed (Tyagi et al., 1998).
Personal protection by the use of repellents is of con-
siderable importance within the integrated disease-vec-
tor control endeavour (Curtis et al., 1989). In this light,
the present study was carried out to evaluate the protec-
tion affordable against Aedes aegypti L. by six formu-
lations of lemongrass oil prepared from different class-
es of base.
j Materials and Methods
Extraction of lemongrass oil
Lemongrass oil was obtained by hydro distillation (B.
P., 1980) from the fresh aerial parts of Cymbopogon
citratus growing within Ile-Ife town. The plant was au-

0944-7113/02/09/03-259 $ 15.00/0
260 A. O. Oyedele et al.
thenticated at the Department of Pharmacognosy
(Obafemi Awolowo University, Ile-Ife, Nigeria)
herbarium.
Preparation of lemongrass oil formulations
Graded concentrations of lemongrass oil (0.25–25%
v/v) in liquid paraffin were prepared. Typical formula-
tions of the oil were also made for repellency testing by
incorporating the requisite quantity of oil into selected
representative formulation bases to obtain stable prod-
ucts containing 15% v/w of the oil. This concentration
of oil was selected based on some preliminary data
(Table 1) and corroborated by an earlier study (Oyedele
et al., 2000). The following bases were investigated:
Oleaginous ointment bases namely, white petrolatum
only, and simple ointment containing cetostearyl alco-
hol (5% w/w), hard paraffin (5% w/w), wool fat (5%
w/w), and white petrolatum (85% w/w) (The Pharma-
ceutical Codex, 1979); surfactant-stabilized cream
bases namely, a glyceryl monostearin cream base con-
taining glyceryl monostearate (5%w/w), sodium lauryl
sulphate (3% w/w), cetostearyl alcohol (2%w/w), liq-
uid paraffin B.P. (25% w/w), and water (to 100%)
(Oyedele et al., 2000), and a cetomacrogol cream base
containing cetomacrogol emulsifying wax B.P. (9%
w/w), liquid paraffin B.P. (6% w/w), white petrolatum
(15% w/w), and water (70% w/w); as well as a hy-
drophilic ointment base containing macrogol 4000
(20% w/w) and macrogol 600 (80% w/w) (Orafidiya
et al., 2001).
Breeding of mosquitoes
Aedes aegypti adult mosquitoes were raised in netted
cages (37 × 30 × 28 cm3) under laboratory conditions
(25–30 °C, R.H. 60–70 %) from the larval colony al-
ready established in our laboratories (Adebayo et al.,
1999). Adult female mosquitoes were fed regularly
with blood from exposed skin of live chicken and the
males with a 10% sugar solution.
Repellency testing
When 80–100 adult mosquitoes had been raised, they
were fed regularly for 4–5 days and then blood-starved
(but water-provided) for the next 48 h before using
them for repellency test. The test sample was applied
thinly to the experimental bird’s bare skin using camel-
hair brush. The bird was then placed against the netted
cage. The rate of biting attack of the test mosquitoes
was determined in triplicate experiments of 2-minute
exposure of the treated bird’s skin to the insects as pre-
viously described (Oyedele et al., 2000). The number
of mosquitoes that bite within 2 min. was counted and
compared to the control bite. (Principles of laboratory
animal care, NIH publication No. 85–23, were com-
plied with.) Repellency was calculated as percent bite
deterrence (± SEM) as previously reported (Oyedele et
al., 2000; Gbolade et al., 2000). This was determined at
hourly intervals until 5 h following application of the
test sample. Results of the tests were compared with
those of the bland formulation bases and that of liquid
paraffin (solvent for the oil) as negative controls, and
against that of citral (Sigma), a major constituent in
lemongrass oil (Ekundayo, 1985). A commercial topi-
cal insect-repellent preparation, Wipe(R), containing di-
ethyl toluamide (20%) and dimethyl phthalate (10%)
(SmithKline Beecham Pharmaceuticals Plc.) was used
as the positive control.
j Results and Discussion
Liquid paraffin solutions of lemongrass oil exhibited
concentration-dependent repellency (Table 1). High
concentrations (20%, 25%) provided complete (100%)
protection lasting 1 h. Lower concentrations (1–15%)
also exhibited total repellency that was short-lived im-
mediately following application of the respective solu-
tions. The 1% and 5% oil concentrations sustained
more than 50% repellency for 3 h while the 10–25%

Table 1. Mosquito repellency of lemongrass oil in liquid paraffin

Time (h) after Bite-deterrence (%) of lemongrass oil solutions

application of
sample Concentration (% v/v) of oil in liquid paraffin Positive control
––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– –––––––––––––––––––
0.25% 0.5% 1% 5% 10% 15% 20% 25% 15% Citral

0 13 ± 4 66 ± 14 100 100 100 100 100 100 100

1 0 14 ± 7 76 ± 8 80 ± 10 96 ± 4 96 ± 4 100 100 85 ± 5
2 0 0 57 ± 4 57 ± 4 92 ± 8 94 ± 6 96 ± 4 96 ± 4 65 ± 5
3 0 0 51 ± 8 53 ± 15 93 ± 5 94 ± 6 94 ± 6 94 ± 6 50 ± 10
5 0 0 30 ± 6 40 ± 35 40 ± 5 41 ± 11 41 ± 18 44 ± 4 35 ± 10
 Mosquito-repellent topical product from Lemongrass
concentrations gave higher protection (> 90% repellen-
cy) over the same period (Table 1). Citral (15% v/v in
liquid paraffin) exhibited repellent effects that were
comparable to those of 5% lemongrass oil (Table 1), in-
dicating that this constituent may play an important
role in the repellent action of the oil. Tyagi et al (1998)
have earlier suggested citronellol and geraniol as other
constituents of the oil that may contribute greatly to its
protective properties.
In earlier studies, essential oils from Hemizygia and
Ocimum species have been found to produce similar
concentration-dependent mosquito repellency. Liquid
paraffin solutions of Ocimum gratissimum L. and O.
basilicum L. oils (1% and higher concentrations) were
shown to exhibit bite-protection (≥70%) for 2–3 h and
resulted in complete protection at the 20% and 35%
concentrations respectively (Gbolade et al., 2000).
Also Hemizygia welwitschii (Rolfe) M. Ashby oil at
5–15% solutions in liquid paraffin was reported to pro-
duce 85–100% antifeedant activity for 4 h (Oyedele et
al., 2000). These solutions were shown to give total re-
pellency at 12% and 15% concentrations, which lasted
2 and 4 h respectively, and this indicates a better activi-
ty compared with lemongrass oil in this present study.
Furthermore, Dua et al (1996) have reported more than
50% protection from the bite of Aedes mosquitoes by
methanolic extracts of Lantana camara L. flowers for
4 h, while the repellency produced by Eucalyptus oil
lasted 5 h (Trigg and Hill, 1996).
Table 2 presents a comparison of the repellent effects
of lemongrass oil (15%v/w) in different formulations.
This concentration of the oil in each preparation pro-
duced an initial total protection, which the hydrophilic
ointment sustained for 1 h. The hydrophilic base best
supported the repellent action of the oil for 2 h.
Throughout the test duration, this ointment demon-
strated the highest repellency potential among the oth-
ers, and was closest to Wipe(R) in activity (Table 2).
Table 2. Mosquito repellency of lemongrass oil formulations
Time (h) after Bite-deterrence (%) of preparations containing 15% v/w oil
application of
sample Oleaginous ointments Surfactant-stabilized creams
––––––––––––––––––––––––– –––––––––––––––––––––––––––––
White Simple Monostearin Cetomacrogol
petrolatum ointment cream
based based
0 100 100 100
1 72 ± 8 96 ± 4 86 ± 8
2 28 ± 12 47 ± 6 42 ± 8
3 0 9±6 29 ± 8
5 0 4±4 14 ± 8
261
However, the 15% lemongrass oil solution (Table 1)
was superior to Wipe(R) in that it sustained higher repel-
lency for longer duration. The surfactant-based creams
were next in efficacy, providing 86–100% protection
for 1 h (Table 2). The oleaginous ointments, which con-
sisted mainly of hydrocarbon constituents, were the
least effective. Their appreciable activity (>70% repel-
lency) also lasted 1 h (Table 2). In general, the formula-
tions in each particular class of base were similar in
their repellent actions. The negative controls exhibited
no repellent activity.
Liquid paraffin solution of Hemizygia oil has been
reported to be a more effective mosquito repellent than
the emulsion formulations containing similar concen-
trations of the oil (Oyedele et al., 2000). Emulsification
of an oil results in entrapment of the dispersed droplets,
which hinders release (Rees and Collett, 1975). Anoth-
er correlation of the properties of formulation base with
activity of preparations of an essential oil showed that a
hydrophilic ointment of lemongrass oil made from a
1:1 w/w blend of macrogol 4000 and macrogol 400
produced greater antibacterial effects than the same
concentration of oil in emulsion products (Onawunmi,
1994). This was attributed to better release from the
base.
The repellent effects of all the products dwindled
with time, as might be expected. The vapour phase-me-
diated repellency of an essential oil fades with time ob-
viously due to diffusion and continuous dilution of the
vapour in the local convectional air currents, resulting
in its gradual reduction and elimination from the vicin-
ity. The observed influence of formulation base types
on product repellency thus indicates that the rate of re-
lease as vapour of the repellent principle of the essen-
tial oil (studied under the same laboratory conditions)
was higher from the hydrophilic ointment preparation
than from the surfactant and hydrocarbon-based
(cream and ointment) products, while the oil solutions
Hydrophilic ointment Positive control
–––––––––––––––––––––– –––––––––––––––––––––––––
Macrogol ointment Commercial
cream Insect-repellent
Wipe(R)
100 100 95 ± 4
87 ± 3 100 84 ± 5
51 ± 6 64 ± 4 74 ± 4
35 ± 6 37 ± 4 55 ± 7
13 ± 6 22 ± 8 43 ± 11
262 A. O. Oyedele et al.
most facilitated the release of the repellent. Volatile oils
diffuse faster from liquid (solutions) than from the
semisolid preparations (Oyedele et al., 2000). The slow
release of the volatile repellent principle of lemongrass
oil from the oleaginous formulations can thus be at-
tributed to the lipophilic affinity of the oil and its con-
stituents for the bases, as well as the viscosity of the
base.
Six different formulations of lemongrass oil have
been evaluated in this study. The mosquito bite-deter-
rent effect of 15%v/w hydrophilic ointment formula-
tion of the oil is very promising for topical use, and the
preparation can be reapplied should a longer effect be
required provided no untoward effect is produced.
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j Address
Dr. A. O. Oyedele, Department of Pharmaceutics,
Obafemi Awolowo University, Ile-Ife Nigeria.
e-mail: aoyedele@oauife.edu.ng