Journal of Ethnopharmacology 106 (2006) 312–316

Neurosedative and muscle-relaxant activities of ethyl

acetate extract of Baphia nitida AFZEL
Olufunmilayo O. Adeyemi a,∗ , Omoniyi K. Yemitan a,b , Adefunmilayo E. Taiwo a
a
Department of Pharmacology, College of Medicine of the University of Lagos, Idi-Araba,
P.M.B. 12003 Lagos, Nigeria
b Department of Pharmacology, Lagos State University College of Medicine, P.M.B. 21266 Ikeja, Lagos, Nigeria

Received 18 August 2005; received in revised form 22 November 2005; accepted 30 November 2005
Available online 3 March 2006

Abstract
The sedative, anxiolytic and muscle-relaxant effects of the ethyl acetate leaf extract of Baphia nitida (BN) was investigated in intact mice,
using the hole-board head-dip test for exploratory behavioural effect, elevated plus maze (EPM) and Y-maze (YM) models of anxiety; chimney,
inclined screen, traction and climbing tests for muscle-relaxant effects. In each of these tests, BN (100–400 mg/kg, p.o.), diazepam (1 mg/kg, i.p.)
or distilled water (10 ml/kg, p.o.) was administered, 30 or 60 min before performing the tests in mice. For exploratory behavioural test, number of
head-dip within 15 min was counted. For EPM and YM tests, the cumulative time spent in open and closed arms was recorded within 5 min. In
the muscle-relaxant tests, mice were subjected to modified models such as chimney, inclined screen, traction and climbing tests. BN produced a
significant (P < 0.05) dose-related decrease in exploratory behaviour in the head-dip test and prolongation of cumulative time spent in open arms
of both EPM and YM. BN did not show any significant effect in the chimney and traction tests, but produced significant, dose-dependent muscle
relaxation in the inclined screen and climbing tests. Furthermore, BN (200–1200 ␮g/ml) non-competitively shifted the curves of acetylcholine
contractions of the toad Rectus abdominis muscle to the right. Oral doses of BN (0.1–20 g/kg) did not produce mortality, but the LD50 when given
intraperitoneally, was 645.65 mg/kg. Results suggest that the leaf extract of Baphia nitida has sedative, anxiolytic and skeletal muscle-relaxant
effects and support its neurosedative use in traditional African medicine.
© 2006 Elsevier Ireland Ltd. All rights reserved.

Keywords: Baphia nitida; Sedative; Anxiolytic; Muscle-relaxant; Climbing test; Rectus abdominis

1. Introduction
Baphia nitida (Papilionaceae) has a wide geographical distri-
bution and appears mainly as a shrub or short tree, with immense
benefits. It has been used to provide shades as well as dyewoods.
In addition, it is richly endowed with a wide range of ethno-
pharmacological benefits hence, has been used by indigenes of
many West African countries for medicinal purposes (Irvine,
1961). Baphia nitida is popularly called camwood, but among
the Yoruba people of West Africa, it is called irosun. Natives
of various countries have used different parts of Baphia nitida
for traditional and medicinal purposes. The Yorubas, among
other uses, use parts of the plant for constipation (Irvine, 1961),
ringworm, sprains and swollen joints, parasitic skin diseases,
∗ Corresponding author. Tel.: +234 803 445 9618; fax: +234 1 5851432.
E-mail address: ooadey@yahoo.com (O.O. Adeyemi).
dressing wounds, ulcers and boils and veneral diseases (Dalziel,
1937).
The leaf extract has been reported to possess a depressant
effect on isolated cardiac tissues (Adeyemi, 1992) and neuro-
muscular blocking effect (Adeyemi and Ogunmakinde, 1991).
In this study, the sedative, anxiolytic and muscle-relaxant activ-
ities of the ethyl acetate leaf extract of Baphia nitida (BN) were
investigated.
2. Materials and methods
2.1. Plant material
Fresh leaves of Baphia nitida were collected from Egbeda
area of Lagos State, Nigeria. The botanical authentication was
carried out by Mr. T.K. Odewo, Senior Superintendent, Forestry
Research Institute of Nigeria (FRIN), where a herbarium

0378-8741/$ – see front matter © 2006 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2005.11.035
 O.O. Adeyemi et al. / Journal of Ethnopharmacology 106 (2006) 312–316
voucher specimen (FHI 106544) was deposited for further ref-
erence. Confirmation was done by Professor J.D. Olowokudejo
of the Botany Department, University of Lagos, Lagos, Nigeria.
2.2. Extract preparation
Fresh leaves of the plant (1 kg) were washed and cut into
pieces. This was subsequently placed in the extractor setup in
which 2 l of ethyl acetate was added to the round bottom flask
of the soxhlet extractor for 24 h. The leaf residue was packed
and weighed. About 750 ml of the solvent was recovered in the
process, giving a dark green residue (yield: 2.9%).
2.3. Animals
Adult albino mice of either sex, maintained on standard
rodent feed and given water ad libitum, and toads were obtained
from the Laboratory Animal Centre of the College of Medicine
of the University of Lagos, Lagos, Nigeria.
2.4. Head-dip test
This consists of a square wooden box containing 16 evenly
spaced holes. After 30 min of administration of BN (100, 200
and 400 mg/kg, per os, p.o.), diazepam (1 mg/ml, intraperitoneal,
i.p) or distilled water (10 ml/kg, p.o.), each mouse was placed
at the centre of the wooden box and the number of times the
mouse dipped its head inside the holes was counted for a period
of 3 min (n = 6) (Dhara et al., 2002).
2.5. Anxiolytic tests
2.5.1. Elevated plus maze
The elevated plus maze (EPM; 30 cm × 6 cm × 6 cm, each
arm) is made of wood, consisting of two open and two closed
arms across at 60 cm above the ground level. Each mouse was
placed in the central position facing an open arm and the cumu-
lative time spent in open or closed arms was recorded for 5 min.
BN (100, 200 and 400 mg/kg, p.o.), diazepam (1 mg/kg, i.p.) or
distilled water (10 ml/kg, p.o.) was administered, 30 or 90 min
before performing the tests in the mice (n = 6) (Yemitan and
Adeyemi, 2003).
2.5.2. Y-maze
BN (100, 200 and 400 mg/kg, p.o.), diazepam (1 mg/kg,
i.p.) or distilled water (10 ml/kg, p.o.) was administered, 30 or
90 min to different groups of mice (n = 6) before being placed
at the centre of a Y-shaped wooden runway (Y-maze, YM;
70 cm × 15 cm × 12 cm) with one of the two arms closed. The
cumulative time spent by a mouse in open or closed arm was
recorded for 5 min (Yemitan and Adeyemi, 2003).
2.6. Muscle-relaxant tests
2.6.1. Climbing tests
This test was adopted from a method by Yemitan and
Adeyemi (2003). Male mice were trained to climb a chain, 50 cm
313
long by placing the fore paws of each one on the free end of the
chain. The chain was suspended from a clamp standing on a lab-
oratory bench (90 cm from ground). A normal mouse grasped the
chain with the fore paws and when allowed to hang free, placed
the two feet on the chain and climbed till it got to a marked
point 2 cm to the top of the chain. Mice, which got to the mark
within 30 s was selected for further tests. Previously screened
male mice were used for the test, 30 min after administration
of BN (100, 200 and 400 mg/kg, p.o.), diazepam (1 mg/kg,
i.p.) or distilled water (10 ml/kg, p.o.) to different groups
(n = 6).
2.6.2. Inclined screen test
Thirty minutes after the administration of BN (100, 200 and
400 mg/kg, p.o.), diazepam (1 mg/kg, i.p.) or distilled water
(10 ml/kg, p.o.), each group of mice (n = 6) was left for 1 h on
a flat, slippery, rectangular glass plate (42 cm × 37 cm) inclined
at 30◦ to the horizontal, to observe for a paralyzing effect severe
enough to cause the mice to slide off the screen (modified from
Randall et al., 1960).
2.6.3. Chimney test
A mouse was introduced at one end and allowed to move to the
other end of a Pyrex glass tube (30 cm long × 3.0 cm diameter),
marked at 20 cm from base. When the animal reached the 20 cm
mark, the tube was moved to the vertical position and immedi-
ately, the mouse tried to climb the tube with a backward move-
ment. The mice, which successfully reached the mark within
30 s, were selected for further testing. The test was repeated
with screened animals 30 min after the administration of BN
(100, 200 and 400 mg/kg, p.o.), diazepam (1 mg/kg, i.p.) or dis-
tilled water (10 ml/kg, p.o.) to different groups (n = 6) (Boissier
et al., 1961).
2.6.4. Traction test
The fore paws of a mouse were placed on a small twisted
wire rigidly supported above a laboratory bench top. Normal
mice grasped the wire with the fore paws and when allowed
to hang free, placed at least one hind foot on the wire within
5 s. Inability to place at least one hind foot marked failure
in the traction test (Rudzik et al., 1973). Previously screened
male mice were used for the test after the administration
of BN (100, 200 and 400 mg/kg, p.o.), diazepam (1 mg/kg,
i.p.) or distilled water (10 ml/kg, p.o.) to different groups
(n = 6).
2.7. In vitro neuromuscular effect
Dose–responses to acetylcholine (0.02–6.0 ␮g/ml) was car-
ried out on an isolated toad Rectus abdominis muscle in an
amphibian ringer solution. Dose–responses to acetylcholine was
later repeated in the presence of each concentration of 200, 600
and 1200 ␮g/ml of BN. Experiments continued until maximum
or reducing responses were obtained for acetylcholine concen-
trations.
314 O.O. Adeyemi et al. / Journal of Ethnopharmacology 106 (2006) 312–316

2.8. Acute toxicity test

Orally administered doses of BN (0.1–20 g/kg) were tested

in mice of both sexes (n = 6, 3 per sex) randomly allotted into
groups. Similarly, another set of mice was randomly allotted
to six groups to which BN (0.1–1.6 g/kg) was given intraperi-
toneally to different groups. Distilled water (10 ml/kg, i.p.) was
administered to control mice. For each group, observations were
made within 24 h for lethality and mortality.

2.9. Statistical analysis

Results are expressed as mean ± S.E.M or percentage. Sta-

tistical analysis of data was done by Student’s t-test. A level of
significance (P < 0.05 or 0.01) was considered for each test.
3. Results
3.1. Exploratory behavioural effect
Fig. 1. Effect of Baphia nitida on exploratory behaviour in mice (head-dip
test). Bars represent the number of head-dip in a hole-board, by mice, 30 and
60 min after treatment with distilled water (10 ml/kg, control), Baphia nitida
(100–400 mg/kg) or diazepam (1 mg/kg). Data are presented as mean ± S.E.M.
(n = 6). * P < 0.05; ** P < 0.01, with respect to control.

Mice pretreated with extract (100 and 200 mg/kg) showed
a significant (P < 0.05) dose-dependent decrease in exploratory
behaviour in the hole-board. A significant (P < 0.01) effect com-
parable to that produced by 1 mg/kg diazepam, was produced by
400 mg/kg of BN (Fig. 1).
in the open arm of Y-maze compared to control, 30 and
60 min post-administration. Effect produced at 400 mg/kg of
extract was comparable (P < 0.01) to that produced by diazepam
(Table 1).

3.4. Climbing tests

3.2. Elevated plus maze
A progressive dose dependent increase in the time taken
Mice pretreated with extract (100, 200 or 400 mg/kg) showed
to climb the wire, was produced 30 and 60 min post-
a significant (P < 0.05) dose-dependent, prolongation of the
administration of BN (100–400 mg/kg). A peak response was
cumulative time spent in the open arms of the elevated plus
produced with 400 mg/kg of extract, which was comparable
maze, 30 and 60 min post-administration relative to control mice.
with diazepam (P < 0.05) at 30 min post-administration of BN
BN (400 mg/kg) produced a peak effect significantly higher
(Fig. 2).
than control (P > 0.01) and diazepam (P < 0.05) at 30 min post-
treatment (Table 1).
3.5. Inclined screen test
3.3. Y-maze
A significant (P < 0.05) reduction in time taken to slide off
The extract (100, 200 and 400 mg/kg) produced a signifi- the inclined screen was produced with 400 mg/kg of the extract
cant dose-dependent prolongation of the cumulative time spent and was comparable to that produced by diazepam (Fig. 3).

Table 1
Effect of Baphia nitida on anxiety (elevated plus maze and Y-maze)
Treatment and dose (mg/kg) Time spent in open arms as a function of total time spent in both open and closed arms (%)

Elevated plus maze Y-maze

30 min 60 min 30 min 60 min

Control 29.4 21.1 22.2 26.5

BN (100) 37.8a,c 39.3b,c 29.4a,c 29.6c
BN (200) 50.0b 43.4b,c 36.7a,c 34.0a
BN (400) 53.4b,c 48.6b 45.2b 37.9a
Diazepam (1) 46.0b 47.5b 50.0b 39.2a

Values represent percentages of time spent in open arms as a function of total time spent in both open and closed arms in the elevated plus maze or Y-maze, 30 or
60 min after treatment with distilled water (10 ml/kg, control), Baphia nitida (100–400 mg/kg) or diazepam (1 mg/kg); Student’s t-test (n = 6).
a P < 0.05.
b P < 0.01, significantly different from control.
c P < 0.05, significantly different from diazepam.
 O.O. Adeyemi et al. / Journal of Ethnopharmacology 106 (2006) 312–316 315

Table 2
Effect of Baphia nitida on muscle relaxation (chimney and traction tests)
Treatment Dose (mg/kg) Percentage of mice showing negative test

Chimney test (%) Traction test (%)

Control – 0 0
BN 100 0 0
BN 200 0 0
BN 400 0 0
Diazepam 1 100 100

Values represent percentages of mice (n = 6) showing negative effects in the

chimney and traction tests, 30 min after treatment with distilled water (10 ml/kg,
control), Baphia nitida (100–400 mg/kg) or diazepam (1 mg/kg).

Fig. 2. Effect of Baphia nitida on muscle tone (climbing test). Bars represent
the time (in s), by mice to climb a suspended chain, 30 and 60 min after treat-
ment with distilled water (10 ml/kg, control), Baphia nitida (100–400 mg/kg)
or diazepam (1 mg/kg). Data are presented as mean ± S.E.M. (n = 6). * P < 0.05,
with respect to control.

3.6. Chimney test

No significant effect was produced in mice pretreated with

BN (100–400 mg/kg) in the time taken to climb backward in
the Pyrex glass tube. However, mice pretreated with diazepam
could not climb backwards in the Pyrex glass tube (Table 2).

3.7. Traction test

Fig. 4. Effect of Baphia nitida (BN) on acetylcholine (Ach)-induced contraction
BN (100–400 mg/kg) did not impair the ability to place at of toad Rectus abdominis muscle. Plots represent dose-related, non-competitive
least one hind foot on the wire within 5 s, unlike diazepam inhibition of Ach responses, produced by BN (200–1200 ␮g/ml). Each point is
(Table 2). the mean of six values ± S.E.M.

3.8. In vitro neuromuscular effect tion of the toad Rectus abdominis muscle as shown in
Fig. 4.
BN (200–1200 ␮g/ml) produced dose-dependent, non-
competitive inhibition of acetylcholine-induced contrac- 3.9. Acute toxicity test

Orally administered doses of BN (0.1–20 g/kg) did not pro-

duce mortality in mice, but the median lethal dose (LD50 ) of
intraperitoneally administered BN was 645.65 mg/kg.

4. Discussion

A dose-dependent suppression of exploratory behaviour in

Fig. 3. Effect of Baphia nitida on muscle tone (inclined screen test). Bars repre-
sent the time (in min), by mice to slide off an inclined screen, 30 min after treat-
ment with distilled water (10 ml/kg, control), Baphia nitida (100–400 mg/kg)
or diazepam (1 mg/kg). Data are presented as mean ± S.E.M. (n = 6). * P < 0.05,
with respect to control.
the head-dip test is an indication that BN produces a central ner-
vous system (CNS) depressant activity (File and Wardill, 1975).
Furthermore, the extract producing a significant dose-
dependent prolongation of cumulative time spent in the open
arms of the EPM and YM, compared with the control, suggests
that BN causes less fear and less anxiety toward open and ele-
vated area (Yemitan and Adeyemi, 2003), and is comparable to
that produced by diazepam, an anti-anxiety drug. In both tests,
a dose of 400 mg/kg of the extract produced very significant
results showing that it acts by an inhibitory effect on the cen-
tral nervous system in a manner, similar to diazepam. The EPM
and YM tests are designed to detect the effect of anxiolytic drugs
(Hogg, 1996; Yemitan and Adeyemi, 2003). The anxiolytic, anti-
316 O.O. Adeyemi et al. / Journal of Ethnopharmacology 106 (2006) 312–316
convulsant, muscle-relaxant and sedative-hypnotic actions of
benzodiazepines make them the most important GABAA modu-
lating drugs (Doble and Martin, 1996; Stafford et al., 2005) and
this may explain the mechanisms of action of BN as well. It has
also been reported that the anxiolytic action of benzodiazepines
such as diazepam, may be mediated by a direct activation of
glycine synapses in the brain (Snyder and Enna, 1975). The
mechanism of anxiolytic action of BN might therefore involve an
action on both gamma aminobutyric acid (GABA) and glycine
neurotransmission; however, further studies are needed to ascer-
tain this. Furthermore, the phytochemical tests on the major
constituents of the ethyl acetate extract of the plant have yielded
a red pigment reported to be santalene (C33 H26 O11 ) (Mertini and
Taylor-David, 1981) as well as flavones, saponins and tannins
(Farnsworth and Euler, 1962; Odebiyi and Sofowora, 1978). It
is possible that the mechanism of anxiolytic, as well as other
CNS depressant actions of BN could be due to the binding of
any of these phytochemicals to the GABAA –benzodiazepine
complex. In support of this, it has been found that flavones,
which are present in the plant, bind with high affinity to the
benzodiazepine site of the GABAA receptor (Kahnberg et al.,
2002).
Experimental results show that the extract produced a signifi-
cant skeletal muscle relaxation as depicted in the inclined screen
and climbing tests which could be comparable with diazepam, a
standard skeletal muscle-relaxant. Locomotor activity is con-
sidered as an index of alertness and a decrease in it would
indicate sedative activity (Thakur and Mengi, 2005). Benzodi-
azepines, like diazepam, have been reported to produce their
muscle-relaxant property by a central action independent of
their sedative effect (Rang and Dale, 1991) and through mim-
icking of glycine at its post-synaptic receptor sites in the CNS
(Young et al., 1974). BN might have similar mechanisms of
action.
The result of the in vitro test indicates that the muscle-relaxant
effect of BN might also involve neuromuscular cholinergic sites.
This latter effect is a confirmation of an earlier report that the
aqueous extract of Baphia nitida, like the ethyl acetate extract
used in the present study, produced a non-competitive inhibi-
tion of acetylcholine-induced contraction of the toad Rectus
abdominis muscle (Adeyemi and Ogunmakinde, 1991). How-
ever, the ethyl acetate extract has shown more potency. There
is therefore an indication that the active muscle-relaxant princi-
ple, though present in the aqueous phase, is more extracted in
ethyl acetate. When administered orally, BN (0.1–10 g/kg) did
not produce mortality up to 14 days post-treatment, indicating
that the extract is quite safe. Results obtained suggest that the
leaf extract of Baphia nitida possesses sedative, anxiolytic and
skeletal muscle-relaxant activities possibly mediated through the
GABAA /benzodiazepine and glycine inhibitory mechanisms.
These results provide a justification for the use of the plant
extract to alleviate anxiety and muscle tension, and support its
use as a psychosedative in traditional African medicine. Further
investigation of the mechanism(s) of action of the plant extract,
as well as the active substance(s) responsible for its biological
actions, is going on in our laboratory.
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