Professional Documents
Culture Documents
PR 08022
PR 08022
Abstract
Purpose: Oxidative stress has been shown to play an important role in the development of anaemia in
malaria. Indeed, increase in total antioxidant status has been shown to be important in recovery from
malaria. The antioxidant activities of four medicinal plants traditionally used in the treatment of malaria
in southwestern Nigeria were determined.
Methods: The ethanolic extracts of the leaves of Carica papaya Linn. [Caricaceae] , stem bark of
Magnifera indica Linn. [Anacardiaceae], leaves of Psidium guajava Linn. [Myrtaceae] and the leaves of
Vernonia amygdalina Del. [Compositae], were used in the present study. The plant parts commonly
used in the locality in malaria therapy were employed in this study. The plants were screened for the
presence of phytochemicals and, their effect on 2,2-Diphenyl-1-picryl-hydrazyl radical (DPPH) was used
to determine their free radical scavenging activity.
Results: Phytochemical screening of the plants showed the presence of flavonoids, terpenoids,
saponins, tannins and reducing sugars. M. indica did not contain cardiac glycosides and alkaloids while,
P. guajava also showed the absence of alkaloids and anthraquinones. Anthraquinones was similarly
absent from V. amygdalina. Concentrations of the plant extracts required for 50% inhibition of DPPH
radical scavenging effect (IC50) were recorded as 0.04 mg/ml, 0.313 mg/ml, 0.58 mg/ml, 2.30 mg/ml and
0.054 mg/ml for P. guajava, M. Indica, C. papaya, V. amygdalina and Vitamin C, respectively.
Conclusion: All the plants showed potent inhibition of DPPH radical scavenging activity, P. guajava
being the most potent. The free radical scavenging (antioxidant) activities of these plants probably
contribute to the effectiveness of the above plants in malaria therapy.
Key words: Carica papaya, Magnifera indica, Psidium guajava, Vernonia amygdalina, Antioxidants,
Malaria, DPPH, Oxidative stress.
ammonia was added. The resulting solution gently to extract the alkaloidal base. The
was observed for colour changes. chloroform layer was extracted with 10 ml of
acetic acid. This was divided into two
Test for terpenoids (Salkowski test)
portions. Mayer’s reagent was added to one
To 0.5 g each of the extract was added 2 ml of portion and Draggendorff’s reagent to the
chloroform. Concentrated H2S04 (3 ml) was other. The formation of a cream (with Mayer’s
carefully added to form a layer. A reddish reagent) or reddish brown precipitate (with
brown colouration of the interface indicates Draggendorff’s reagent) was regarded as
the presence of terpenoids. positive for the presence of alkaloids.
Test for flavonoids Test for cardiac glycosides (Keller-Killiani
test)
Three methods were used to test for
flavonoids. First, dilute ammonia (5 ml) was To 0.5 g of extract diluted to 5 ml in water was
added to a portion of an aqueous filtrate of the added 2 ml of glacial acetic acid containing
extract. Concentrated sulphuric acid (1 ml) one drop of ferric chloride solution. This was
was added. A yellow colouration that underlayed with 1 ml of concentrated
disappear on standing indicates the presence sulphuric acid. A brown ring at the interface
of flavonoids. Second, a few drops of 1% indicated the presence of a deoxysugar
aluminium solution were added to a portion of characteristic of cardenolides. A violet ring
may appear below the brown ring, while in the
the filtrate. A yellow colouration indicates the acetic acid layer a greenish ring may form just
presence of flavonoids. Third, a portion of the above the brown ring and gradually spread
extract was heated with 10 ml of ethyl acetate throughout this layer.
over a steam bath for 3 min. The mixture was
filtered and 4 ml of the filtrate was shaken with Determination of antioxidant activity
1 ml of dilute ammonia solution. A yellow
The radical scavenging activities of the plant
colouration indicates the presence of
extracts against 2,2-Diphenyl-1-picryl hydrazyl
flavonoids.
radical (Sigma-Aldrich) were determined by
Test for saponins UV spectrophotometry at 517 nm. Radical
scavenging activity was measured by a
To 0.5 g of extract was added 5 ml of distilled slightly modified method previously
water in a test tube. The solution was shaken described12,13. The following concentrations
vigourously.and observed for a stable of the extracts were prepared, 0.05, 0.1, 0.5,
persistent froth. The frothing was mixed with 1.0, 2.0 and 5 mg/ml in methanol (Analar
3 drops of olive oil and shaken vigourously grade). Vitamins C was used as the
after which it was observed for the formation antioxidant standard at concentrations of 0.02,
of an emulsion. 0.05, 0.1, 0.2, 0.5 and 0.75 mg/ml. I ml of the
Test for tannins extract was placed in a test tube, and 3 ml of
methanol was added followed by 0.5 ml of 1
About 0.5 g of the extract was boiled in 10 ml mM DPPH in methanol. A blank solution was
of water in a test tube and then filtered. A few prepared containing the same amount of
drops of 0.1% ferric chloride was added and methanol and DPPH. The radical scavenging
observed for brownish green or a blue-black activity was calculated using the following
colouration formula:
Test for alkaloids % inhibition = {[Ab-Aa]/Ab} x 100 ……(1)
0.5 g of extract was diluted to 10 ml with acid where Ab is the absorption of the blank
alcohol, boiled and filtered. To 5 ml of the sample and Aa is the absorption of the extract
filtrate was added 2 ml of dilute ammonia. 5
ml of chloroform was added and shaken
Reducing sugar + + + +
Anthraquinone + - - -
Terpenoids + + + +
Flavonoids + + + +
Saponins + + + +
Tannins + + + +
Alkaloids + - + -
Cardiac + - + +
glycosides
100
90
% Inhibition of DPPH
80
70
60
50
40
30
20
10
0
0.05 0.1 0.5 1 2 5
Concentration mg/ml
5. Das B.S, Nanada NK. Evidence for erythrocyte lipid 1993; pp 150.
th
peroxidation in acute falciparum malaria. 10. Trease G.E., and Evans, W.C. Pharmacognosy. 13
Trans. Roy. Soc. Trop. Med. Hyg 1999; 93: 58- edn. Bailliere Tindall, London, 1989, pp 176-
62. 180.
6. Kremsner PG, Greve B, Lell B, Luckner D and 11. Harbone JB. Phytochemical Methods London.
Schmidt D. Malarial anaemia in African Chapman and Hall Ltd, 1973, pp49-188.
Children associated with high oxygen-radical 12. Brand-Williams W, Cuvelier ME, Berset C. Use of
production. Lancet 2000; 355: 40-41. free radical method to evaluate antioxidant
7. Loria P, Miller S, Foley M, Tilley L. Inhibition of activity. Lebensmittel Wissenschaft und
peroxidative degradation of heme as the basis Technologie 1995; 28: 25-30.
of action of chloroquine and other quinoline 13. Ayoola GA, Sofidiya T, Odukoya O, Coker H.A.B
antimalarials. Biochem. J 1999; 339: 363-370. Phytochemical screening and free radical
8. Odugbemi TO, Odunayo RA, Aibinu IE, Fabeku, OP. scavenging activity of some Nigerian medicinal
Medicinal plants useful for malaria therapy in plants. J. Pharm. Sci. & Pharm. Pract. 2006; 8:
Okeigbo, Ondo State, Southwest Nigeria. Afr. J. 133-136.
Trad. CAM. 2007; 4(2): 191-198. 14. Polterait O. Antioxidants and free-radical scavengers
9. Sofowora A. Medicinal plants and Traditional of Natural Origin. Current Org. Chem. 1997; 1:
Medicine in Africa. Spectrum Books, Ibadan. 415-440.