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Journal of Medicinal Plants Research Vol. 4(11), pp. 991-994, 4 June, 2010
Available online at http://www.academicjournals.org/JMPR
ISSN 1996-0875 ©2010 Academic Journals
Full Length Research Paper
In vitro anti-malarial activity of 20 quinones isolated
from four plants used by traditional healers in the
Democratic Republic of Congo
J. S. Kayembe*, K. M. Taba, K. Ntumba, M. T. C. Tshiongo and T. K. Kazadi
*Department of Chemistry, Faculty of Sciences, University of Kinshasa, Democratic Republic of Congo, P. O. Box 190,
Kinshasa XI.
Accepted 28 January, 2010
The anti-malarial activity of 20 quinones isolated from Cassia alata, Cassia occidentalis, Garcinia kola
and Ocimum basilicum was investigated in vitro using the micro dilution test of Desjardin by a visual
evaluation on thin blood smears. The six quinones isolated from C. occidentalis, three from C. alata and
three from O. basilicum were found to be the most active with an IC50 value of below 1 µg/ml. The others
quinones showed a moderate activity with IC50 values of between 5 and 20 µg/ml.
Key words: Cassia alata, Cassia occidentalis, Garcinia kola, Ocimum basilicum, quinones, anti-malarial
activity, in vitro.
INTRODUCTION
Malaria accounts for 1 - 3 million deaths yearly
worldwide, with most of this burden occurring in children
under 5 years of age in sub - Saharan Africa (Breman
2001). In Kinshasa, Democratic Republic of the Congo,
malaria is prevalent in pupils in suburbs where most of
poor people live (Kazadi et al., 2004). An ethno botanical
survey carried out in these areas had shown that more
than 70% of populations use plants to treat fever
(supposed to be malaria) (Ngalamulume et al., 1995). 58
species of plants and recipes commonly used were
identified. Among those, Cassia alata, Cassia
occidentalis, Garcinia. kola and Ocimum basillicum were
the most frequently used (Kasuku et al., 1999).
The intolerable burden of malaria can be conquered by,
among other things, searching new molecules with potent
anti - malarial activity in plants which always have been
the most common sources of medicines.
The aim of the present work is to evaluate in vitro anti -
malarial activities of 20 quinones isolated from these four
plants: C. alata (4), C. occidentalis (6), G. kola seeds (5)
and O. basillicum (5). In the chemotherapy of malaria,
most molecules belong to the class of alkaloids (Federici
et al., 2000; Andrade – Neto et al., 2003; Zirihi et al.,
*Corresponding author. E-mail: jeanksm@unikin.cd.
2005) and terpenes (Li et al., 1992). The class of
quinones has few representative such as Atovaquone
(Fowler et al., 1994; Basco et al., 1995), Lapachol and
Lapinone (The Merck Index 2005). Quinones as
antimalarial are attracting attention recently
(Kanokmedhakul et al., 2005; Laurent et al., 2006; Eyong
et al., 2006) and the present investigation is in line with
this interest.
MATERIALS AND METHODS
Plant materials
Plants were collected in Kinshasa/ Kisenso, Democratic Republic of
Congo and were authenticated by the Herbarium service of
Department of Biology, University of Kinshasa where voucher
specimens are preserved. The leaves and seeds were air dried at
room temperature for 20 days and then grinded with pestle and
mortar.
Preparation of crude ethanol extract
Crude ethanol extracts were prepared for primary in vitro anti-
malarial assay. The dried and grinded leaves or seeds (20 g) were
extracted with ethanol (2 ×1 L) at room temperature. After the
removal of ethanol in vacuo, the crude extracts obtained were
submitted for anti-malarial test in vitro.
992 J. Med. Plant. Res.
Table 1. In vitro antimalarial activity of crude ethanol extracts.
Plant Concentration ( µg/ml) Growth inhibition %
Cassia alata
Cassia occidentalis
Garcinia kola
Ocimum basilicum
Quinine
25
12.5
25
12.5
n.d
12.5
0.1
83
53
93
24
n.d
78
100
Extraction of quinones
The dried and grinded leaves (200 g) were extracted with methanol
(2 x 2.5 l) at room temperature. The methanol extract was
suspended in a Methanol - H2O 7/3 mixture and then extracted
successively with Petroleum ether (60 - 80°C) and CHCl3.The
CHCl3 fraction was subjected to vacuum liquid chromatography
(VLC) on silica gel previously soaked in a 0.5N oxalic acid solution
and dried over night. The fractions having the same TLC profile
were collected, concentrated together and then submitted each to
column chromatography on silica gel using the following solvent
systems: Pyridine/Amyl alcohol/H2O 4 : 6 : 3 for C. alata quinones;
CHCl3/Ethyl Acetate 5 :1 for C. occidentalis; Ethyl Acetate/
Petroleum Ether 4 :1.2 for G. kola and O. basillicum quinones. Four
fractions (CA1 - CA4) for C. alata, six fractions (CO1 - CO6) for C.
occidentalis, five fractions (GK1 - GK5) for G. kola and five fractions
(OB1 - OB5) for O. basillicum were obtained. All these fractions
showed a positive Borntrager test with a 5% KOH alcoholic
solution.
Anti - malarial activity
The anti - malarial activities of quinones were evaluated in vitro on
their ability to inhibit Plasmodium falciparum growth on RPMI 1640
medium. This method estimates the growth of parasites incubated
in RPMI 1640 medium containing plants extracts in various
concentrations (Desjardins et al., 1979). Parasitemia was
determined on blood smears 48 h after extracts and parasites
contact (Benoit et al., 1996). The IC50 values were determined
graphically in terms of concentration versus inhibition percentage.
RESULTS AND DISCUSSION
Ethanol extracts
Screening the anti - malarial activity of crude extracts is a
first step in the isolation of new molecules with potent
activity (Ma, Zhang and al. 2006; Njoroge G.N. and
al.2006). Anti - malarial activities results of crude ethanol
extracts (Table 1) show that for a concentration of 25
µg/ml, C. occidentalis extract is the most active with 93%
of plasmodium growth inhibition compared to the others
crude extracts. C. occidentalis extracts have been
reported to possess various biological activities (Sharma
et al., 2000; Bin-Hafeez et al., 2001). The anti – malarial
activities of C. occidentalis and G. kola have been
reported by Tona et al. (2001). It is likely that these
activities could be attributed to the presence of quinones
in these plants. The Petroleum Ether and ethanol extracts
of C. alata are reported to have a potent inhibitory activity
against Chrysomya megacephale (Kumarasinghe et al.,
2002) and against opportunistic HIV patients’ infections
(Crockett et al., 1992). Ocimum genus species are known
to have antimicrobial, antispasmodic and mosquitoes
repellent effects (Fatope he results obtained for the 20
quinones are summarized in (Table 2). 12 quinones (CO1
- CO6; CA1, CA2 and CA4; OB1, OB2 and OB4) were found
to be the most active with the IC50 values below 1 µg/ml.
4 quinones showed an acceptable activity with the IC50
values between 1 and 5.5 µ/ml. The others have shown a
low activity (IC50 > 10 µg/ml).
Natural quinones have benzoquinone, naphtoquinone
or anthraquinone structures. A growing interest is granted
to these compounds from the description of the
antimalarial activity of Atovaquone (Basco et al., 1995).
Three quinones isolated from Salacia krausii presented
very high activity on P. falciparum (Figueiredo et al.,
1998). Weiss et al. reported a good in vitro antimalarial
activity for Naphthoquinones isolated from Kigelia pinnata
with IC50 values around 0.002 µg/ml (Weiss et al., 2000).
This naphtoquinone is more active than all isolated by us.
A new Xestoquinone isolated in Vanuatu from a marine
sponge (Xestospongia sp.) exhibited also an interesting
inhibitory activity on plasmodium growth with an IC50
value of 3 µM (Laurent et al., 2006). These results added
to ours, showed that quinones are compounds with an
interesting antimalarial activity. They could be used as
starting point for the synthesis of molecules much more
active such as the reported synthesis of an
aminonaphtoquinone which has shown a IC50 of 37,3
ng/ml (Kapadia et al., 2001).
Conclusion
Quinones isolated from C. alata, C. occidentalis, G. kola
and O. basilicum have interesting antimalarial activities
(IC50 < 1 µg/ml for 12 of them). Further investigations are
required in order to characterize the isolated compounds.
Kayembe et al. 993
Table 2. In vitro IC50 (µg/ml) of isolated quinones against Plasmodium falciparum.
Plant Isolated quinone Rf IC
50
(µg/ml)
Cassia alata (leaves)
CA
1
0.61 <0.1
CA
2
0.70 5.4
CA
3
0.86 0.54
CA4
0.92
<0.25
Cassia occidentalis (leaves)
CO
1
0.17 0.25
CO
2
0.28 0.25
CO
3
0.33 <1
CO
4
0.53 0.25
CO
5
0.64 <0.1
CO6
0.84
<0.1
Garcinia kola (seeds)
GK
1
0.17 1.02
GK
2
0.25 2.0
GK
3
0.42 12.9
GK
4
0.54 15.75
GK5
0.85
n.d
Ocimum basilicum (leaves)
OB
1
0.12 0.52
OB
2
0.23 <0.35
OB
3
0.50 1.42
OB
4
0.63 <0.35
OB5
0.88
18
Naphthoquinones (Kigelia pinnata)* 0.002
Aminonaphtoquinones** 37.3 ng/ml**
Chloroquine
72 ng/ml**
*Weiss et al. (2000).
**Reported by Kapadia et al. (2001).
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