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Acute and sub-chronic oral toxicity studies of

an aqueous stem bark extract of Pterocarpus
soyauxii Taub (Papilionaceae) in rodents

ARTICLE in JOURNAL OF ETHNOPHARMACOLOGY · OCTOBER 2010

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 Journal of Ethnopharmacology 133 (2011) 329–335

Contents lists available at ScienceDirect

Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jethpharm

Acute and sub-chronic oral toxicity studies of an aqueous stem bark extract of
Pterocarpus soyauxii Taub (Papilionaceae) in rodents
M.C. Tchamadeu a,d,e , P.D.D. Dzeufiet a , P. Nana a , C.C. Kouambou Nouga a , F. Ngueguim Tsofack a ,
J. Allard d,e,f , N. Blaes d,e , R. Siagat c , L. Zapfack b , J.P. Girolami d,e,∗ , I. Tack d,e,f , P. Kamtchouing a , T. Dimo a
a
Department of Animal Biology and Physiology, Faculty of Science, University of Yaounde 1, P.O. Box 812, Yaounde, Cameroon
b
Department of Plant Biology and Physiology, Faculty of Science, University of Yaounde 1, P.O. Box 812, Yaounde, Cameroon
c
Department of Organic Chemistry, Faculty of Science, University of Yaounde 1, P.O. Box 812, Yaounde, Cameroon
d
INSERM, U858, F-31432 Toulouse Cedex 4, France
e
Institut de Médecine Moléculaire de Rangueil, University of Toulouse, UPS, IFR31, F 31432 Toulouse Cedex 4, France
f
Department of Physiology, University of Toulouse, UPS, 133, Route de Narbonne, F-31062 Toulouse, France

a r t i c l e i n f o a b s t r a c t

Article history: Pterocarpus soyauxii Taub (Papilionaceae) is used in Cameroonian traditional medicine and pharma-
Received 18 May 2010 copoeia to treat hypertension, diabetes, gastrointestinal parasitizes and cutaneous diseases.
Received in revised form Aim of the study: The present investigation was carried out to evaluate the safety of an aqueous stem bark
14 September 2010
extract of Pterocarpus soyauxii by determining toxicity after acute and sub-chronic oral administration in
Accepted 27 September 2010
male and female rodents.
Available online 13 October 2010
Materials and methods: The acute toxicity test was conducted in mice. An aqueous extract of barks was
administrated by gavage in single doses of 2.5–12.5 g/kg. General behaviour and mortality were examined
Keywords:
Pterocarpus soyauxii
for up to 7 days. The sub-chronic toxicity test was performed in rats. The plant extract was adminis-
Aqueous stem bark extract tered by daily gavage of 150–600 mg/kg for 42 days. Body weight, food and water intakes were followed
Acute and sub-chronic toxicity weekly. Haematological, biochemical and organ parameters were determined at the end of the 42-day
Mortality administration.
Blood analysis Results: In the acute study in mice, oral administration of the aqueous extract of Pterocarpus soyauxii
Histopathology caused dose-dependent general behaviour adverse effects and mortality. The no-observed adverse effect
level (NOAEL) of the extract was 5.0 g/kg. The lowest-observed adverse effect level (LOAEL) was 7.5 mg/kg.
Mortality increased with the dose, LD50 was > 10.75 g/kg for the mouse. In the sub-chronic study in rats,
daily oral administration of the aqueous extract of Pterocarpus soyauxii did not result in death or signifi-
cant changes in haematological or biochemical parameters, excepted increased hepatic catalase activity
(P < 0.05) at the dose of 600 mg/kg. No alteration was observed in body weight, food and water intake.
Liver, kidney, lung and pancreas histopathology did not reveal morphological alteration.
Conclusions: The results showed that the aqueous stem bark extract of Pterocarpus soyauxii Taub had
very low toxicity in oral acute high dose administration and no toxicity in oral sub-chronic low dose
administration and indicate that the plant could be considered safe for oral medication.
© 2010 Elsevier Ireland Ltd. All rights reserved.

1. Introduction
Several species of the genus Pterocarpus disseminated in tropical
regions are widely used in Africa and Asia to treat diseases. Ptero-
carpus soyauxii is a 30–55-feet high rain forest tree. It belongs to the
Fabaceae or Papilionaceae family, branch of spermaphytes (Burkill,
1995). The plant stem bark, grey-brown to brown-coloured, scales
∗ Corresponding author at: INSERM, U858, BP 84225, 31432 Toulouse Cedex 4,
France. Tel.: +33 5 61 32 26 21.
E-mail addresses: marieclaire tchamadeu@yahoo.fr (M.C. Tchamadeu),
Jean-Pierre.Girolami@inserm.fr (J.P. Girolami).
off in fine irregular scales and contains a red sap. Leaves, wood,
stem bark, seed and flours are used in African traditional medicine,
especially in the Cameroonian pharmacopoeia, for treating various
diseases including hypertension, diabetes, intestinal parasitizes,
renal and cutaneous diseases. Bark is used as diuretic in Gabon
and, fresh leaves are also used as food in Nigeria (Oteng-Gyang and
Mbachu, 1987; Kimpouni, 1999; Okafor, 1999; Sarah, 1999).
Pterocarpus soyauxii contains various compounds such as
biflavonoids (santalin A, santarubins A and B), isoflavonoids (pte-
rocarpin, formononetin and prunetin), an isoflavone quinone
(claussequinone), isoflavanes (vestitol and mucronulatol), tannins,
ascorbic acid, glucosides, triterpenes, xanthones (Arnone et al.,
1977; Banerjee and Mukherjee, 1981; Bezuidenhoudt et al., 1987;

0378-8741/$ – see front matter © 2010 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2010.09.035
330 M.C. Tchamadeu et al. / Journal of Ethnopharmacology 133 (2011) 329–335
Kiec-Swierczynska et al., 2004; Surowiec et al., 2004). Other phe-
nolic constituents have been isolated from Pterocarpus marsupium
Roxb bark and Pterocarpus santalinus plant (Manickam et al., 1997;
Kesari et al., 2004; Maurya et al., 2004).
There is a lack of experimental reports on the pharmacologi-
cal activities of Pterocarpus soyauxii extracts. However, Pterocarpus
soyauxii presents similarities in phytochemical composition with
other species of the genus Pterocarpus which biological activities
have been tested. Several components are known for their antidi-
abetic potential (Jung et al., 2006). Roxb-(−)-epicatechin, a water
soluble benzopyran, stimulates insulin secretion (Hii and Howell,
1984). In addition, the biological activities of crude extracts of
Pterocarpus species include hypoglycaemic and antidiabetic prop-
erties (Pterocarpus marsupium (Abesundara et al., 2004; Saxena
and Vikram, 2004; Vats et al., 2004; Anandharajan et al., 2005;
Dhanabal et al., 2006), Pterocarpus santalinus (Nagaraju et al., 1991;
Kameswara Rao et al., 2001)) and also, cardiotonic activity (Pte-
rocarpus marsupium (Mohire et al., 2007)), antimicrobial activity
(Pterocarpus santalinus (Narayan et al., 2007); Pterocarpus angolen-
sis (Steenkamp et al., 2004; Sami et al., 2009); Pterocarpus indicus
(Ragasa et al., 2005)), antiulcerogenic activity (Pterocarpus san-
talinus (Narayan et al., 2007)). Methanolic extracts of Pterocarpus
santalinus have also been reported to induce apoptosis of cancer
HeLa cells (Kwon et al., 2006).
Despite knowledge of biological activities of the Pterocarpus
genus, toxicological studies are very few. Pterocarpus soyauxii wood
dust is known to cause skin irritation, allergic contact dermati-
tis and sensitization (Kiec-Swierczynska et al., 2004). Surprisingly,
although its oral use is widespread, the oral toxicity of Pterocarpus
soyauxii has not been studied.
Therefore, acute and sub-chronic toxicological studies were
conducted in mice and rats, respectively, to evaluate the safety of
the oral administration of an aqueous stem bark extract of Ptero-
carpus soyauxii.
2. Materials and methods
2.1. List of abbreviations
Alanine aminotransferase (ALT), serum aspartate aminotrans-
ferase (AST), malondialdehyde (MDA), catalase (CAT), body weight
(BW).
2.2. Plant material and preparation of the aqueous extract
Pterocarpus soyauxii (Papilionaceae) barks were collected in
March 2002 in Nkolbibanda village (Central province, Cameroon)
by Dr. L. Zapfack (Botany Department, University of Yaounde 1). The
plant was identified at the National Herbarium of Yaoundé where a
voucher specimen was deposited (No.: HNC/2427). The stem barks
were dried at room temperature and ground into powder. Dry pow-
der (200 g) was macerated in 2 l of boiling distilled water for 10 min
and then kept 24 h at room temperature. The resulting aqueous
extract was filtered. The filtrate was concentrated in a drying-room
at 40 ◦ C for 24 h, producing 24.10 g of red well-dried residue (w/w
yield: 12.05%). The extract was stored at −20 ◦ C.
2.3. Phytochemical screening
To determine the chemical constituents, qualitative phyto-
chemical screening of the aqueous stem bark extract of Pterocarpus
soyauxii was carried out following standard procedures routinely
used in the laboratory (Tchamadeu et al., 2010) and revealed alka-
loids (Meyer and Dragendoff’s test), tannins (FeCl3 test), saponins
(frothing test), lipids (Wattman paper test), flavonoids (Schinoda’s
test), glycosides and polyoses (NaCl, and Fehling’s solutions A and
B), anthraquinones (ether–chloroform and NaOH), phenols (FeCl3 ),
polyphenols (K3 Fe(CN)6 ), terpenoids (Liberman Burchard’s test).
2.4. Animals
BALB/c mice and Wistar rats, raised in the animal core facil-
ity of the Faculty of Science, University of Yaoundé 1, were used.
Animals were housed in colony cages (5 rats or mice per cage),
under standard laboratory conditions (ventilated room, 24 ◦ C, 75%
humidity, 12 h light/dark cycle) and had free access to standard
commercial diet and tap water. All animal experiments were con-
ducted in accordance with the internationally accepted principles
for laboratory animal use and care as described in the European
Community guidelines (Official Journal of European Union L197 vol.
50, July 2007).
2.5. Acute oral toxicity study in mice
Healthy BALB/c mice (3-month old, 19–30 g BW) were randomly
assigned to each of six groups of 10 mice (5 females and 5 males).
Mice were fasted overnight (12 h) with free access to water prior to
administration of single doses (0.0, 2.5, 5.0, 7.5, 10.0 and 12.5 g/kg)
of the extract dissolved in distilled water. Treatment was given by
gavage of 1 ml/100 g rat BW or of 0.5 ml/20 g mouse BW. The general
behaviour of the mice was continuously monitored for 4 h after
the treatment, intermittently during a 24-h period (Twaij et al.,
1983), and thereafter daily up to 7 days. The LD50 was determined
as previously described (Molle, 1986).
2.6. Sub-chronic oral toxicity study in rats
Healthy Wistar rats (3-month old, 145–265 g BW) were ran-
domly assigned to each of four groups of 10 rats (5 females and 5
males). The extract, dissolved in distilled water, was administered
by daily gavage for 42 days, to groups I to IV (doses of 0, 150, 300
and 600 mg/kg, respectively). The animals were observed for signs
of toxicity and mortality throughout the experimental period. The
BW, water and food consumption were recorded weekly. At the end
of the 42-day experiment, the animals, fasted for 12 h, were sac-
rificed by decapitation under anaesthesia (thiopental 50 mg/kg).
Blood was collected into two tubes: tube 1 containing EDTA was
processed immediately for haematological parameters; tube 2
without additive was centrifuged at 3000 × g at 4 ◦ C for 10 min to
obtain serum (stored at −20 ◦ C until analysis). The organs (kidneys,
liver, lungs, heart, testes and glands annexes, ovaries, spleen and
pancreas) were weighted. Organ samples (kidney, pancreas, lung
and liver) were either fixed in 10% formalin for histopathologi-
cal examination or stored at −20 ◦ C until analysis of tissue total
proteins, MDA, glutathione and catalase activities.
2.6.1. Blood analysis
Blood glucose was determined on days 0 and 42, using test strips
(Accuchek, Roche Diagnostics), on 20 ␮l caudal vein blood sam-
ples obtained from fasted rats. Whole blood cell (WBC), red blood
cell (RBC), leukocyte and platelet counts, hematocrit, haemoglobin,
mean corpuscular volume (MCV), mean corpuscular haemoglobin
(MCH) and mean corpuscular haemoglobin concentration (MCHC),
were determined as previously described (Tan et al., 2008), using an
automatic analyser (System H1, Bayer Diagnostics). The serum was
analysed as previously described (Tchamadeu et al., 2010) for total
protein, cholesterol, creatinine, triglycerides and (ALT/AST) activi-
ties, using specific commercial diagnostic kits (Fortress Diagnostics,
London, UK). Glutathione level was determined enzymatically
(Ellman, 1959). The optical density at the appropriate wavelength
was measured using a spectrophotometer (UV spectrophotometer
Hitachi).
 M.C. Tchamadeu et al. / Journal of Ethnopharmacology 133 (2011) 329–335
2.6.2. Kidney and liver samples biochemistry
Homogenates of kidney and liver samples were prepared in
Tris–KCl buffer (pH 7.4). After grinding, the mixture was cen-
trifuged at 12,000 × g, 4 ◦ C for 20 min, and the homogenates were
divided into four samples, stored at −20 ◦ C until analysis for tissue
total proteins, MDA (thiobarbituric acid reactive species method),
glutathione (enzymatic method, Ellman, 1959) and catalase activity
(Sinha, 1972). Optical density was monitored using a spectropho-
tometer (UV spectrophotometer Hitachi).
2.6.3. Histopathological analysis
Samples of liver, kidney, lungs and pancreas were fixed in 10%
neutral formalin and processed by conventional techniques. Paraf-
fin sections (5 ␮m thick) were stained with haematoxylin and eosin
before microscopic examination.
2.7. Statistical analysis
Data are presented as mean ± S.E.M. One-way ANOVA with
Dunett’s Multiple Comparison post-test was performed to assess
differences between groups (Graphpad Software, San Diego, CA,
USA). Values of P < 0.05 were considered statistically significant.
3. Results
3.1. Phytochemical screening of the aqueous extract of
Pterocarpus soyauxii
The phytochemical screening of the aqueous stem bark extract
of Pterocarpus soyauxii revealed the presence of the following
classes of chemical compounds: alkaloids, saponins, flavonoids,
tannins, terpenoids, phenols, polyphenols, glucids like glucose;
lipid components were absent.
3.2. Acute oral toxicity of Pterocarpus soyauxii stem bark extract
in mice
The effects of oral administration of single doses of aqueous Pte-
rocarpus soyauxii extract in mice are summarized in Table 1. There
was a regular dose-dependent increase in mortality and adverse
effects in both sexes. Males were more sensitive than females.
Twelve, i.e. 8 males and 4 females, of the 50 treated mice died.
The mortality rate (0% at 5.0 g/kg) progressively rose to 70% at
the highest dose tested (12.5 g/kg). The no-observed-adverse-effect
level (NOAEL) for the oral dose extract was 5.0 g/kg. The lowest-
observed-adverse-effect level (LOAEL) (Alexeeff et al., 2002) was
7.5 g/kg. The symptoms of toxicity were similar in both sexes.
Symptoms such as hair bristling or piloerection, body writhing,
hind limbs and body stretching, occurred within 15–120 min after
the oral administration. Diarrhoea, gasping and breathing difficulty,
immobility or constant changing position to diminish pain, and
weight loss (results not shown) were observed later and at high
doses. Diarrhoea occurred for males before females. The acute tox-
icity data indicated that the estimated oral LD50 of the aqueous
extract of Pterocarpus soyauxii was >10.75 g/kg in mice.
3.3. Sub-chronic oral toxicity of Pterocarpus soyauxii stem bark
extract in rats
3.3.1. General behaviour and mortality
Daily oral administration of Pterocarpus soyauxii extract for 42
days did not induce any obvious symptom of toxicity in rats of
both sexes, included the highest dose tested of 600 mg/kg. No
lethality was recorded for any dose during the 42 days of plant
administration. No differences in general behaviour, food and water
consumptions (Table 2) were observed between the groups of rats.
331
Changes in body weight during the treatment period are presented
in Table 3. As expected, rats gained weight with time. No difference
in body weight gain was noted between the control and the treated
groups.
3.3.2. Haematological and biochemical parameters
The effect of sub-chronic administration of Pterocarpus soy-
auxii on haematological parameters is presented in Table 4. All the
parameters remained within the physiological range throughout
the 42-day experimental period.
Biochemical parameter profiles of the treated and control
groups are shown in Table 5. A 42-day oral administration of
aqueous stem bark extract of Pterocarpus soyauxii did not cause
significant changes in serum creatinine, cholesterol, glutathione
and the transaminase activities (ALT/AST). Triglycerides were lower
(P < 0.05) in the groups II and IV. However, the effect was not dose-
dependent since it was not observed in the group III. The serum
glucose level was lower at D42 compared to D0 in the group IV but
a decrease at D42 was also noted in the control group I. The data on
the renal and hepatic glutathione, MDA and catalase are presented
in Table 5. No statistical difference was detected excepted a signif-
icant (P < 0.05) increase in hepatic catalase activity at the dose of
600 mg/kg, as compared with the control group I.
3.3.3. Organ parameters
Absolute and relative organ weights of 42-day treated rats are
shown in Table 6. There was no significant change in the organ
weights of the treated compared to the control groups. Histopatho-
logical analysis of liver, kidney, lung and pancreas did not show
morphological alteration (Data not presented).
4. Discussion and conclusion
Respiratory and dermatological reactions, due to sawdust
inhalation and direct skin contact, respectively, of Pterocarpus soy-
auxii wood have been reported (Kiec-Swierczynska et al., 2004) to
inform users about the risks or unsafety of wood manipulation.
Although wood, stem bark, leaves and roots of Pterocarpus soyauxii
are used as drug in medicine or as vegetable in cooking, experi-
mental studies referring to the risks of the oral use of the plant
and pharmacological studies are lacking. The present investigation
was carried out to estimate safety limits of oral administration of
the aqueous stem bark extract of Pterocarpus soyauxii Taub (Papil-
ionaceae) through toxicological assessment in rodents.
Estimation of safety of drugs and plant products is currently per-
formed in animals. A good correlation has been reported between
toxicological insults in rats and humans, correlation is weaker
between humans and mice (Olson et al., 2000). Therefore numerous
studies investigate the acute effects of high doses in mice and the
chronic effects of lower doses in rats including the doses potentially
usable in humans (Rhiouani et al., 2008; Lahlou et al., 2008).
In the oral acute toxicity study, mice administered doses up to
5.0 g/kg did not exhibit any sign of adverse effect (NOAEL). Higher
doses induced mortality and symptoms of pronounced adverse
effects. The LD50 for the oral administration of the Pterocarpus soy-
auxii aqueous extract was estimated to be >10.75 g/kg BW of mice,
a dose much higher than the medicinal dosage. Thus, referring to
the Hodge and Stemer scale (CCOHS, 2005), the orally adminis-
tered stem bark aqueous extract of Pterocarpus soyauxii could be
considered practically non-toxic or at worst slightly toxic.
A gender difference in acute mortality of aqueous Pterocarpus
soyauxii extract was observed in mice. A gender effect has been
observed for other plant extracts, the difference in susceptibility
being, as here for Pterocarpus soyauxii, in a narrow range of doses.
For example, the female were more sensitive to plant extracts
332 M.C. Tchamadeu et al. / Journal of Ethnopharmacology 133 (2011) 329–335

Table 1
Acute oral toxicity of an aqueous extract of Pterocarpus soyauxii Taub in mice.

Treatment dose (g/kg) Mice Effects

Sex D/T Mortality latency (h) Symptoms of toxicity

0.0 M 0/5 None

F 0/5
2.5 M 0/5 None
F 0/5
5.0 M 0/5 None
F 0/5
7.5 M 1/5 >6, <7 Piloerection, trembling, fore-limbs and body stretching
F 0/5
10.0 M 3/5 >5, <48 Piloerection, trembling, fore-limbs and body stretching, wriggling,
F 1/5 >7, <10 diarrhoea, breathing difficulty, constant changing position, immobility.
12.5 M 4/5 >4, <48 Piloerection, trembling, fore-limbs and body stretching, diarrhoea, gasping
F 3/5 >5, <48 or breathing difficulty, asthenia, constant changing position, immobility.

The aqueous stem bark extract of Pterocarpus soyauxii, dissolved in distilled water was administered as single oral doses to 6 groups of 10 mice (5 males, 5 females). All
animals were carefully examined for adverse effects (behavioural changes and mortality) for 48 h. Symptoms of toxicity are described for a group (both males and females).
M: male; F: female; D/T: dead/treated mice; latency: time to death after the dose; none: no toxic symptom during the observation period.

Table 2
Effect of sub-chronic oral administration of an aqueous extract of Pterocarpus soyauxii Taub on food and water intakes in rats.

Week of treatment Treatment

Group I (0 mg/kg) Group II (150 mg/kg) Group III (300 mg/kg) Group IV (600 mg/kg)

Food (g/day/rat) (n = 10)

W0 24.06 ± 4.13 27.32 ± 2.74 26.63 ± 4.51 25.96 ± 3.12
W1 21.38 ± 2.45 21.32 ± 3.09 19.65 ± 2.60 21.52 ± 2.68
W2 23.58 ± 3.95 21.93 ± 3.94 22.23 ± 3.18 22.58 ± 2.95
W3 16.03 ± 1.62 17.47 ± 2.53 17.58 ± 1.93 16.30 ± 0.87
W4 18.83 ± 1.19 22.98 ± 2.97 24.23 ± 4.17 19.20 ± 1.67
W5 19.58 ± 1.28 20.82 ± 4.58 21.25 ± 7.13 19.14 ± 2.51
W6 19.29 ± 0.28 22.84 ± 1.93 21.97 ± 2.74 20.30 ± 3.42
Water (ml/day/rat) (n = 10)
W0 21.40 ± 2.00 25.80 ± 4.60 25.70 ± 1.10 25.40 ± 0.80
W1 29.50 ± 4.00 29.85 ± 5.85 30.25 ± 0.75 35.03 ± 3.93
W2 29.94 ± 6.39 33.15 ± 6.75 30.80 ± 5.00 32.90 ± 4.70
W3 28.60 ± 5.20 30.45 ± 5.65 35.57 ± 0.27 31.38 ± 3.38
W4 20.30 ± 0.70 27.73 ± 1.63 33.18 ± 0.03 28.20 ± 4.60
W5 20.10 ± 0.10 22.10 ± 2.10 31.20 ± 5.00 25.70 ± 5.50
W6 27.10 ± 2.90 27.60 ± 3.60 33.50 ± 1.10 31.80 ± 0.40

An aqueous Pterocarpus soyauxii extract was administered by daily gavage for 42 days to groups of 10 rats (5 males, 5 females) at the doses of: 0, 150, 300 and 600 mg/kg.
Food and water consumptions were evaluated weekly (7 days) and expressed as g/day/rat and ml/day/rat, respectively. Data are mean ± S.E.M. W0: week before treatment.
There were no significant differences, both in food and water consumptions, between the groups of rats at any time period.

of Artemisia afra (Mukinda and Syce, 2007) or Tanacetum vulgare
(Lahlou et al., 2008).
In the sub-chronic toxicity study in rats, aqueous Pterocar-
pus soyauxii stem bark extract was administered at doses of
150–600 mg/kg BW. There were no significant changes in animal
behaviour, food and water consumptions as well as in body weight
gain. A decrease in body weight would be an indicator of adverse
effects (Raza et al., 2002; Teo et al., 2002).
Analysis of blood parameters is relevant to risk evaluation of
alterations of the haematological system in humans (Olson et al.,
2000). No significant alterations of the haematological and bio-
chemical parameters of both male and female treated rats can be
attributed to the plant extract. A slight decrease in blood glucose
level was observed at the highest dose tested. Since a decrease in
blood glucose was also observed in the control group I at the end
of the 42-day experiment, this rules out an hypoglycaemic effect

Table 3
Effect of sub-chronic oral administration of an aqueous extract of Pterocarpus soyauxii Taub on the body weight of rats.

Week (W) of treatment (n = 10) Treatment

Group I (0 mg/kg) Group II (150 mg/kg) Group III (300 mg/kg) Group IV (600 mg/kg)

Body weight (g) (n = 10)

W0 195.0 ± 25.6 195.3 ± 35.2 196.4 ± 17.2 192.3 ± 26.4
W1 208.8 ± 28.6 214.9 ± 43.4** 207.3 ± 25.4 208.9 ± 32.5
W2 225.1 ± 27.5** 224.7 ± 44.8** 219.8 ± 29.2** 223.9 ± 36.9**
W3 240.3 ± 30.3** 239.1 ± 45.9** 233.5 ± 38.8** 241.2 ± 45.8**
W4 240.4 ± 30.7** 244.7 ± 42.6** 237.7 ± 41.7** 241.8 ± 19.8**
W5 250.6 ± 32.7** 250.7 ± 36.2** 246.1 ± 45.9** 248.4 ± 13.8**
W6 256.1 ± 32.9** 259.8 ± 42.0** 253.6 ± 44.4** 253.0 ± 14.0**

Wistar rats (n = 10) were administered an aqueous extract of Pterocarpus soyauxii by daily gavage for 42 days. Data are expressed as mean ± S.E.M.
**
P < 0.01 vs the corresponding pre-treatment value (W0) of each group. There were no significant differences in the body weights between the groups of rats at any time
period.
 M.C. Tchamadeu et al. / Journal of Ethnopharmacology 133 (2011) 329–335 333

Table 4
Effect of sub-chronic oral administration of an aqueous extract of Pterocarpus soyauxii Taub on haematological parameters of rats.

Parameter Treatment

Group I (0 mg/kg) Group II (150 mg/kg) Group III (300 mg/kg) Group IV (600 mg/kg) Normal range

Male (n = 5)
WBC (×103 /␮l) 8.54 ± 1.35 11.20 ± 0.65 10.14 ± 1.95 9.86 ± 0.6 6.5–11.5
Lymphocytes (×103 /␮l) 6.92 ± 0.75 8.36 ± 0.75 8.52 ± 0.60 7.86 ± 0.81 5.8–9.2
RBC (×106 /␮l) 8.99 ± 0.96 9.02 ± 0.09 9.07 ± 0.01 9.17 ± 0.6 6.4–9.5
Haemoglobin (g/dl) 17.14 ± 0.65 16.40 ± 0.15 17.08 ± 0.35 16.78 ± 0.95 13–18
Hematocrit (vol.%) 45.54 ± 1.00 44.64 ± 0.15 44.90 ± 0.10 47.34 ± 4.95 31–54
Platelets (×103 /␮l) 565.0 ± 14.50 498.60 ± 2.5 582.8 ± 1.50 557.8 ± 21.5 450–750
MCV (fl) 50.74 ± 1.45 49.88 ± 0.65 49.54 ± 0.10 51.60 ± 2.05 48.5–55.0
MCH (pg) 19.14 ± 0.70 18.44 ± 0.00 18.84 ± 0.40 18.32 ± 0.20 17.5–22.0
MCHC (g/dl) 37.68 ± 1.30 37.02 ± 0.04 38.10 ± 0.75 35.52 ± 1.95 32.0–43.0
Female (n = 5)
WBC (×103 /␮l) 9.10 ± 0.79 10.84 ± 1.03 8.44 ± 0.87 8.58 ± 0.89 6.5–11.5
Lymphocytes (×103 /␮l) 8.76 ± 0.40 6.66 ± 0.65 6.58 ± 0.35 7.1 ± 0.65 5.8–9.2
RBC (×106 /␮l) 8.57 ± 0.16 7.88 ± 0.16 7.79 ± 0.34 7.95 ± 0.38 6.4–9.5
Haemoglobin (g/dl) 16.30 ± 0.12 16.30 ± 0.24 15.88 ± 0.28 16.28 ± 0.32 13–18
Hematocrit (vol.%) 43.26 ± 0.75 40.74 ± 0.77 39.96 ± 1.87 40.92 ± 1.87 31–54
Platelets (×103 /␮l) 641.4 ± 24.64 696.6 ± 41.12 654.6 ± 20.3 655.80 ± 81.83 450–750
MCV (fl) 50.54 ± 0.50 51.66 ± 0.24 51.32 ± 0.45 51.56 ± 0.83 48.5–55.0
MCH (pg) 19.04 ± 0.33 20.70 ± 0.41 20.48 ± 0.62 20.64 ± 0.85 17.5–22.0
MCHC (g/dl) 37.68 ± 0.75 40.12 ± 0.85 40.02 ± 1.26 40.04 ± 1.33 32.0–43.0

Wistar rats (n = 10 per group, 5 males and 5 females) were administered an extract of Pterocarpus soyauxii by daily gavage for 42 days. Data are expressed as mean ± S.E.M.
No significant difference in any haematological parameter was observed between the groups of rats.

Table 5
Effect of sub-chronic oral administration of an aqueous extract of Pterocarpus soyauxii Taub on biochemical parameters of rats.

Parameter Treatment

Group I (0 mg/kg) Group II (150 mg/kg) Group III (300 mg/kg) Group IV (600 mg/kg) Normal range

Serum (n = 10)
Glucose (mg/dl) D0 115.00 ± 1.50 100.20 ± 2.50 110.40 ± 2.00 113.30 ± 2.0 80.00–120.00
D42 93.20 ± 5.01a 107.60 ± 2.50 96.10 ± 2.50 86.50 ± 3.01 aa
Total cholesterol (mg/dl) 66.08 ± 2.77 78.23 ± 4.42 68.73 ± 2.21 75.25 ± 1.66 60.00–80.00
Triglycerides (mg/dl) 96.96 ± 7.97 71.16 ± 5.08* 86.09 ± 7.16 72.61 ± 7.25* 70.00–100.00
ALT (U/l) 98.98 ± 3.06 95.92 ± 3.06 83.88 ± 6.12 109.18 ± 4.08 80.00–120.00
AST (U/l) 37.64 ± 0.20 34.80 ± 0.60 41.92 ± 0.80 45.92 ± 1.20 30.00–50.00
Serum creatinine (mg/l) 3.27 ± 0.46 3.20 ± 0.12 3.48 ± 0.34 3.48 ± 0.23 3.00–3.70
Serum protein (g/l) 109.22 ± 6.91 101.15 ± 2.30 103.23 ± 2.30 98.39 ± 2.30 3.00–3.70
Serum glutathione (mmol/ml) 0.56 ± 0.19 0.40 ± 0.07 0.44 ± 0.17 0.50 ± 0.10 0.40–0.60
Liver (n = 10)
Hepatic protein (g/l) 61.52 ± 3.46 61.29 ± 1.15 71.66 ± 2.30 66.82 ± 1.15 60.00–75.00
Hepatic glutathione (mmol/g) 3.79 ± 0.18 3.60 ± 0.04 3.97 ± 0.13 4.04 ± 0.07 3.50–4.20
Hepatic MDA (nmol/g) 9.33 ± 0.53 10.29 ± 0.50 10.17 ± 0.61 8.91 ± 0.27 8.00–12.00
Hepatic catalase (U/min/mg) 6.08 ± 2.25 8.45 ± 2.75 8.42 ± 1.58 10.32 ± 2.75* 6.00–10.00
Kidney (n = 10)
Renal protein (g/l) 52.53 ± 2.30 46.54 ± 3.46 48.85 ± 3.46 53.69 ± 1.15 45.00–55.00
Renal glutathione (mmol/g) 3.98 ± 0.57 4.29 ± 0.48 3.92 ± 0.11 4.20 ± 0.31 3.80–4.30
Renal MDA (nmol/g) 4.69 ± 2.18 4.00 ± 0.03 4.29 ± 0.72 4.29 ± 0.75 4.00–4.80
Renal catalase (U/min/mg) 10.28 ± 0.33 9.97 ± 0.50 10.15 ± 0.25 11.12 ± 1.08 9.00–12.00

Wistar rats (n = 10 per group) were administered an aqueous extract of Pterocarpus soyauxii by daily gavage for 42 days. Data are expressed as mean ± S.E.M.
a
P < 0.05 vs day 0.
aa
P < 0.01 vs day 0.
*
P < 0.05 vs the corresponding Group I value.

of the plant extract. Of note, other species of the genus Pterocarpus
such as Pterocarpus marsupium (Jung et al., 2006) or Pterocarpus san-
talinus (Kameswara Rao et al., 2001) exhibit hypoglycaemic activity.
Triglycerides were reduced in some groups (II and IV), but the effect
was not dose-dependent and thus cannot be attributed to the plant
extract.
There were no adverse effects on usual markers of liver and
kidney toxicity, i.e. serum levels of ALT/AST and creatinine, respec-
tively. Furthermore, there were no changes in kidney levels of
MDA, glutathione and catalase activity. These results, associated to
the observed normal creatinine levels suggest that the stem bark
extract of Pterocarpus soyauxii did not alter kidney function at the
doses studied. For the liver, an increase in hepatic antioxydative
catalase activity was observed, at the highest dose tested. Since the
MDA (end products of lipid peroxydation) and antioxydative glu-
tathione levels did not change, this increase should not be indicative
of liver alteration. No difference was observed in the weight and
structure of liver, kidney, lung and pancreas between the control
and the treated groups. Altogether, the sub-chronic study indi-
cates that Pterocarpus soyauxii ingestion did not induce detrimental
changes and morphological alterations in these organs.
In conclusion, the present investigation provides valuable infor-
mation on the acute and sub-chronic toxicity profiles of oral use
of aqueous stem bark extract of Pterocarpus soyauxii. Information
will help for future clinical studies of the medicinal safety and in
vivo experimental studies of the pharmacological potentialities of
this mode of administration of the plant medicine. The aqueous
stem bark extract of Pterocarpus soyauxii, when administered orally,
presented a dose-dependent toxicity profile. Toxicity was achieved
with acute high doses in mice. At lower doses, the extract appeared
334 M.C. Tchamadeu et al. / Journal of Ethnopharmacology 133 (2011) 329–335

Table 6
Effect of sub-chronic oral administration of an aqueous extract of Pterocarpus soyauxii Taub on organ weights of rats.

Organ (g) Organ weight (g) Relative organ weight (g per 100 g body weight)

Group I Group II Group III Group IV Group I Group II Group III Group IV
(0 mg/kg) (150 mg/kg) (300 mg/kg) (600 mg/kg) (0 mg/kg) (150 mg/kg) (300 mg/kg) (600 mg/kg)

Male (n = 5)
Kidneys 0.79 ± 0.04 0.77 ± 0.02 0.83 ± 0.04 0.78 ± 0.03 0.27 ± 0.01 0.26 ± 0.02 0.28 ± 0.01 0.27 ± 0.03
Testicles 1.42 ± 0.11 1.37 ± 0.11 1.40 ± 0.09 1.38 ± 0.03 0.48 ± 0.04 0.46 ± 0.05 0.48 ± 0.03 0.48 ± 0.02
Epididyms 0.44 ± 0.04 0.43 ± 0.06 0.45 ± 0.09 0.47 ± 0.01 0.15 ± 0.01 0.14 ± 0.02 0.15 ± 0.03 0.16 ± 0.01
Liver 6.90 ± 0.63 7.21 ± 0.24 7.21 ± 1.01 7.09 ± 0.54 2.33 ± 0.20 2.40 ± 0.16 2.46 ± 0.34 2.43 ± 0.02
Heart 0.77 ± 0.03 0.82 ± 0.03 0.84 ± 0.10 0.81 ± 0.03 0.26 ± 0.01 0.27 ± 0.00 0.29 ± 0.03 0.28 ± 0.01
Lungs 1.54 ± 0.08 1.38 ± 0.02 1.78 ± 0.12 1.62 ± 0.03 0.52 ± 0.03 0.46 ± 0.02 0.61 ± 0.05 0.56 ± 0.03
Pancreas 0.39 ± 0.03 0.40 ± 0.02 0.44 ± 0.03 0.36 ± 0.05 0.13 ± 0.01 0.13 ± 0.01 0.15 ± 0.01 0.12 ± 0.01
Spleen 0.58 ± 0.07 0.60 ± 0.03 0.57 ± 0.10 0.60 ± 0.03 0.20 ± 0.02 0.20 ± 0.01 0.20 ± 0.03 0.21 ± 0.02
Female (n = 5)
Kidneys 0.56 ± 0.01 0.62 ± 0.02 0.62 ± 0.06 0.58 ± 0.08 0.26 ± 0.01 0.28 ± 0.01 0.29 ± 0.02 0.27 ± 0.01
Ovaries 0.08 ± 0.01 0.08 ± 0.01 0.08 ± 0.01 0.07 ± 0.02 0.04 ± 0.01 0.03 ± 0.01 0.04 ± 0.01 0.03 ± 0.01
Liver 5.83 ± 0.47 5.96 ± 0.22 5.90 ± 0.39 5.62 ± 0.39 2.70 ± 0.18 2.72 ± 0.10 2.77 ± 0.06 2.64 ± 0.09
Heart 0.57 ± 0.11 0.59 ± 0.03 0.66 ± 0.04 0.61 ± 0.03 0.26 ± 0.05 0.27 ± 0.01 0.31 ± 0.01 0.29 ± 0.02
Lungs 1.17 ± 0.04 1.19 ± 0.05 1.41 ± 0.11 1.22 ± 0.10 0.54 ± 0.01 0.55 ± 0.02 0.66 ± 0.5 0.57 ± 0.04
Pancreas 0.37 ± 0.06 0.37 ± 0.03 0.35 ± 0.01 0.42 ± 0.03 0.17 ± 0.02 0.17 ± 0.01 0.16 ± 0.01 0.20 ± 0.01
Spleen 0.47 ± 0.06 0.54 ± 0.02 0.49 ± 0.06 0.47 ± 0.02 0.22 ± 0.02 0.24 ± 0.01 0.23 ± 0.02 0.22 ± 0.01

Wistar rats (n = 10, 5 males and 5 females per group) were administered an aqueous extract of Pterocarpus soyauxii by daily gavage for 42 days. Data are expressed as
mean ± S.E.M. There was no significant difference.

non-toxic in mice and rats, up to the doses of 5.0 g/kg and 600 mg/kg
(the highest tested), respectively. Thus, at the doses empirically
used in traditional medicine, the plant, at least its aqueous stem
bark extract, could be considered with a wide margin of safety for
oral use. Since toxicity in humans cannot always be entirely extrap-
olated from animal studies, clinical evaluation should be performed
to precisely define the safe dosage to advise in humans.
Acknowledgements
This work was supported by funds from the Third World
Academy of Sciences (Grant No. 06-020RG/BIO/AF/AC-UNESCOFR:
3240144852), the French-Cameroon Cooperation (French Embassy
in Cameroon) and the National Institute of Health and Medical
Research–Institute of Molecular Medicine of Rangueil (INSERM
U858-I2MR), Toulouse, France. The authors thank C. Pecher for
technical assistance.
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