Abstract
The development
of new drugs imperative is made by the problem of resistance of malarial
parasites to available anti-malarial drugs .Discovery of new drugs is an
alternative source with the use of natural plant products. The aim of this is
to evaluate the anti-malarial activity of eluted fractions of Acacia nilotica root extract and also
the phytochemicals responsible for this activity. The gradient of solvent
mixture such as hexane, ethyl acetate and methanol were used in multiples of
100 ml where the extract was eluted successively and then each fractions were
collected separately.4 main fractions namely F1, F2,F3 and F4 were produced
which were selected on the basis of elutes that showed similar Rf values and
thin chromatographic profiles. Curative test was used to test those fractions
separately for their anti-malarial activity. The packed cell volume (PCV), temperature
and changes in body weight were also recorded. The results that were obtained
were that fraction F1 at 50 and 100 mg/kg b/w produced significant and dose
dependent reduction in the parasite count of Plasmodium berghei infected mice compared to that of the control
used, there was a significant increment in the survival time of the mice
compared to the control group and the malarial-induced anemia was made better
by improving PCV by Fraction F1 in treated mice. The anti-malarial activity of A.nilotica’s extract was localized in F1
fraction of the extract. The F1 fraction was found to be rich in alkaloids and
phenolics.
INTRODUCTION
Malaria is a
disease caused by a parasite which is preventable and curable but it causes
thousands of infections and death hence it is also called tropical disease. The
resistance of plasmodium parasite to the used anti-malarial drugs is the main
challenge to the effective diagnosed malarial cases. Acacia nilotica which is also known as prickly tree or gum Arabic
tree is a thorny, nitrogen fixing tree that grows to 14-17 cm and 2-3 diameter
and is found in Egypt, South Africa, Kenya and Nigeria. It is used to treat
cough, diarrhea, dysentery and leprosy and to cure impotence. The fruit is used
to treat tuberculosis and powdered pods were used to treat diabetes mellitus.
A.nilotica is rich in secondary metabolites such as tannins, terpenes,
alkaloids, flavonoids and phenolic with known pharmacological properties.
MATERIALS AND METHODS
At first, the
plant was collected, identified and authenticated. The Swiss albino mice (Mus
musculus) were used for the experiment. The parasite used for this experiment
was chloroquine sensitive rodent Plasmodium;
P. berghei. Chloroquine phosphate and silica gel (70-230 mesh) were used as
the main chemicals and other chemicals were prepared using distilled water. The
root extract was air dried and pulverized using grinding machine and the powder
was kept in air tight container in cool and dry place. Aqueous extraction was
prepared by cold maceration process. Here, 400 g of powered root of A.nilotica was soaked in 1 L of
distilled water and kept for 24 hours and was filtered. The filtrate was
freeze-dried and this was carried out 3 times. Wet silica gel was loaded in
column and the extract was kept in upper layer and was eluted in gradients of
solvent mixture in multiples of 100 and the fractions were collected
separately. Elutes having similar Rf values and thin chromatographic profiles
were combined and 4 fractions were made which were tested for anti-malarial
activity each using curative test. The fractions were then subjected to
phytochemical screening to determine the predominant secondary metabolite in
each fraction. The parasite was then inoculated with 0.2 ml of infected blood
in each mouse and then the mice were divided in 5 groups randomly with 6
mice/group. Group 1 was administered 10 mg/kg distilled water (negative
control) , group2,3 and 4 were administered 25,50 and 100 mg/kg b/w of each
fraction and group 5 was administered 5mg/kg b/w chloroquine (positive
control). Same was done for F2, F3 and F4. In established malarial infection,
the rane test was used to evaluate the schizonticidal activity of the fractions
of A.nilotica and mean survival time
was calculated and also percentage parasitemia was determined.
PCV was then
determined using a standard Micro-Hematocrit Reader, the body weight and
temperatures were also monitored. The data so obtained was analyzed by Graphpad
Prism version 5.02 and were expressed as means ±standard error mean (SEM).
DISCUSSION
Chloroquine was
used in this research as a control group because Plasmodium parasite used for this
is chloroquine-sensitive strain. This study was carried out in-vivo to see the
involvement of immune system and effects of prodrug in the removal of infection
in living host. Here, F1 and F2 caused dose-dependent and significant reduction
in mean parasite count in doses 50 and 100mg/kg.
RESULT
From the
experiment, it is seen that fraction F1 shows the anti-malarial activity which
may be due to high proportion of alkaloid found in this fraction or due to
synergistic effect of alkaloids with phenolic compound. This F1 fraction was
able to improve PCV of treated mice either by prevention of malaria-induced
hemolysis or by enhancing erythropoises. But this could not reverse the body
temperature and weight loss associated with rodent malaria.
REFERENCE
Alli LA,
Adesokan AA and Salawu AO (2016) Antimalarial Activity of Fractions of Aqueous
Extract of Acacia nilotica root, Journal of Intercultural Ethnopharmacology, 5 (2); 180-185.
By: Safalta Ranabhat
Fifth Semester, B.Pharm.
School of Health and Allied Sciences
Pokhara University, Dhungepatan, Lekhnath,
Kaski, Nepal.
Tags:
Natural products