Professional Documents
Culture Documents
ISSN: 2041-0778
© M axwell Scientific Organization, 2010
Submitted Date: January 26, 2010 Accepted Date: February 08, 2010 Published Date: March 10, 2010
Abstract: Selected plants (8 species) having a history of use in Sudanese traditional medicine for the treatment
of infectious diseases were investigated for antibacterial activity in vitro. Phytochemical screening of these
plants was performed for constituents: alkaloids, flavonoids, tannins, anthraquinone s, saponins and volatile oils.
Moisture, ash, crude fibres and soluble ethanol extractive contents have been carried out. The antibacterial
screening of the ethanol extracts of the selected plants was performed by the agar well diffusion method against
clinical isolates Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis) and Gram-negative
bacteria (Escherichia co li and Pseudom onas aeruginosa). All the eight ethano lic extracts show ed go od ac tivity
against four tested bacteria. The activity of the Cymbopogon schoenathus spp. proximus aerial parts,
Cymbopogon nervatus inflorescence and Cassia oc ciden talis seed extracts w ere more pronounced. T he results
of the antibacterial activity screening support the ethno medical uses of these plants. Further studies on the
isolation and characterization of the Cymbopogon schoenathus spp. proximus and Cymbopogon nervatus,
partially o r totally responsible for th e observed antibacterial pro perties in progress.
Key w ords: Antibacterial activity, ethanolic extracts, medicinal plants, phytochemical screening, Sudan
Corresponding Author: Dr. Hatil Hashim EL-Kamali, Department of Botany, Faculty of Science and Technology,
Omdurman Islamic University, P.O. Box 382, Omdurman, Sudan
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Curr. Res. J. Biol. Sci., 2(2): 143-146, 2010
Table 2: Antibacterial activity of the ethanolic extracts of selected medicinal plants from central Sudan
Na lidixic
Bac teria A.m G.a C.o C.sn C.t R.m C.n C.s Ge ntam icin Tetracycline acid Am picillin Strep tom ycin Colistin Nitro furan toin
Staphylococcus 12 16 14 12 12 16 15 20 15 7 0 0 9 0 12
aureus
Bac illus sub tilis 15 22 17 13 17 15 20 24 16 18 13 0 16 0 19
Esch erichia coli 15 13 15 15 14 13 14 14 0 15 13 0 10 13 9
Pseudomonas 15 13 16 0 13 14 16 15 15 0 0 0 10 13 0
aeruginosa
A.m = A m b ro si a m a ri ti m a; G.a = Ge igeria a lata; C.o = Cas sia occ identa lis; C.sn = Cassia senna; C.t = Cassia tora; R.m = R h yn ch o si a m i ni m a var. me mn onia ;
C.n = Cymbopogon nervatus; C.s = Cymbopogon schoenanthus ssp. proximus. Values are inhibition zon e (IZ ) dia me ters/m m; 0 = N o inh ibitio n.; Ethanol alone did not show
any activities
Table 3: Properties and phytochemical screening of selected medicinal plants from central Sudan
Plants Plant part Et OH extract Phytochemical M o is tu re (% ) T ota l a sh (% ) A cid in so lu ble as h ( %) S ulf ate d a sh (% )
% yield screening
A m b ro si a m a ri ti m a Leaves 1.5 Alkaloids, Flavonoids, 20.0 20.0 10.0 2.5
Vo latile oil
Ge igeria a lata Ae rial parts 1.8 Alkaloids, Flavonoids, 20.0 25.0 10.0 1.5
Tan nins, V olatile oil
Cas sia occ identa lis Seeds 1.3 Anthraquinones, 15.0 5.0 1.5 3.5
Tannins
Cassia senna Pods 2.5 Anthraquinones, 15.0 2.5 1.5 2.1
Tannins
Cassia tora Fermented 2.6 Anthraquinones, 25.0 20.0 7.0 3.3
leaves Tannins
R h yn ch o si a m i ni m a Roo ts 5.0 Saponins, 5.0 15.0 4.5 2.4
var. me mn onia Anthraquinones
Cymbopogon nervatus Inflorescence 2.0 Flavonoids, Tannins, 25.0 5.0 7.0 4.4
Vo latile oil
Cymbopogon Ae rial parts 1.3 Flavonoids, Tannins, 20.0 10.0 8.0 4.8
schoenanthus ssp. Vo latile oil
proximus
and the standa rd reference antibiotics (Gentamicin-25 mg, incubated in the upright position at 37 ºC for 24h. The
Tetracycline – 25 m g, Nalidixic acid – 30 mg , Am picillin plates were checked for bacterial growth after the
– 25 mg, Streptomycin – 25 mg, Colistin – 25 mg and incubation period and the resultant zon es of growth
Nitrofurantoin – 25 mg) were used. inhibition were accurately measured an d exp ressed in
mm.
Antibacterial testing: The inoculum size of each test
bacteria was adjusted to a 1 ml of bacterial suspension. Properties of selected sudanese med icinal plants: The
Using a flamed and then cooled 9mm. cork-borer, five moisture, ash, acid-insoluble ash, sulfated ash, crude fiber
cups were cut out of each of the ag ar plates wh ich were and alcohol-soluble extractive of selected Sudanese
previously inoculated with the test organism. T he cut medicinal plants (A. maritima, C.alata, C. occidentalis,
discs of agar were removed and decontaminated. By C. senna, C. tora, R. minima var. memnonia,
means of standard pipettes, 0.1ml of each of the different
Cymbopogon nervatus and C. schoenanthus spp.
test suspensions was added to the appropriate cups.
proximus) were determined a ccording to stan dard
Concentration of 25 mg / ml was tested for each plant
procedu res (AO AC , 1980; Lo u, 1980).
extract. Each test was replicated twice. In each plate one
well at the middle was saturated only with the solvent
(ethan ol) and used as a negative control. After filling the Phytochemical tests: Phytochemical screening was
reservoirs with the appropriate dilutions, the plates w ere carried out acco rding to Farnsworth (19 66).
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Curr. Res. J. Biol. Sci., 2(2): 143-146, 2010
Extraction procedures: Dried and powdered plant obtained by hydrodistillation for 4 hours in a C levenger-
materials (100 g) were extracted with 80% ethanol (500 type apparatus (BP, 1980).
ml) in Soxhlet Apparatus for 6 h. The solvent was
removed in vaccum (R otava por R E) to give concentrated RESULTS AND DISCUSSION
extract and the residue kept in the refrigerator until use.
In the present work medicinal uses and the yields of
Tests: ethan olic extracts of candidate plant sp. were
Alkaloids: The ethanolic extract (30 ml) was evaporated phytochemically screened and tested for their antibacterial
to dryness in an eva porating dish on w ater bath. Five ml activity. The results are shown in Tables 1-3. The
of 2NHC l were adde d and stirred w hile heating on the ethanolic extracts of all the examined plants showed
water bath for 10 min., cooled, filtered and the filtrate was stronger grow th inhibition against Gram-positive bacteria.
treated with a few drops of Mayer reagent. The samples Study on antibacterial activity of 8 p lants brought to
were then observed for the presence of turbidity or light some very interesting results. C. nervatus ssp.
precipitation. proximus, C. nervatus and Cassia oc ciden talis appeared
to be the most active against tested bacteria. For
Flavonoids: The alcohol extract (75 m l) of plant samp le Cymbopogon species this effect might be due to the high
were evaporated to dryness on a water bath, cooled and content of terpenoids w hereas for C. occidentalis the
the residue was defatted by washing several times w ith activity might be related to the presence of tannins.
petroleum ether. The defatted residue was dissolved in 30 A glance at Table 2 reveals that various plant
ml 80% ethanol and filtered. The filtrate wa s treated with ethan olic extracts may be bro adly d ivided into two
a few drops of concentrated HCl and magnesium turnings categories (1), those having a relatively higher propensity
(0.5 g). The prese nce of flavonoids was ind icative if a to act on Gram-positive bacteria (2), those equally or
pink or magenta-red colour developed within 3 min.
nearly equally effective against both types of bacteria.
Ethanolic extracts of G. alata, C. schoenanthus ssp.
Tannins: The alcoholic extract (25 ml) was evaporated to
proximus showed relatively higher propensity to act on
dryness on a water bath. The residue was extracted
Gram- positive bacteria. EtOH extracts of C. oc ciden talis,
several times with n-hexane and filtered, the insoluble
C. tora, C. senna, R. minima var. memnonia, A. maritima
residue was stirred with 10 ml of hot saline solution, the
and C. nervatus showed equal or nearly equal
mixture was cooled, filtered and the volume of filtrate
antibacterial activity both against Gram-p ositive and
was adjusted to 10 ml with more saline solution. To 5 ml
Gram-negative bacteria. Compared to the standard,
of this solution, few drops of ferric chloride test reagent
C. occidentalis, C. nervatus, C. schoenanthus ssp.
were added. An intense green, purple, blue or black
proximus ethanolic extracts exhibited a broader spectrum
colour was taken as an evidence for the presence of
of antibacterial activity.
tannins.
EL-Magboul et al. (1985) stated that chloroformic
Saponins: One gram of ethanol extract was dissolved in extracts of C. schoenanthus ssp. proximus was foun d to
10 ml of distilled water in a test tube and shaked be weak activity against S. aureus, B. sub tilis, E. coli and
vigorously for 1-2 min. The presence of saponins was P. aeruginosa except methano lic extrac t against S.
indicated by ch aracteristic hon eyco mb fro th at least 1 cm aureus. However, in the present stud y ethanolic extract of
in height, which persisted for 30 min. this plant was highly effective against all tested bacteria.
The obtained results may provide a support to some uses
Anthraquinone glycosides: To 1 g of the powdered plant of the plants in traditional medicine.
material, 10 ml of N/2 potassium hydroxide containing 1
ml of 3% hydrogen peroxide solution was added. The CONCLUSION
suspension was boiled for 3-5 min. then cooled, filtered
and 5 ml of the filtrate was acidified with 10 drops of Compared to reference antibiotics, the spectrum of
glacial acetic acid. This acidified mixture was extracted antibacterial activity of most investgated plants was found
by shaking with 10 ml of benzene. A 5 ml aliquot of the to be clearly superior. The demonstration of broad
benzene solution was shaken with 3 ml of 10% spectrum of C. schoenanthus ssp. proximus, C. nervatus
ammonium hydroxide solution an d the two layers were and C. occid entalis may help to discover new chemical
allowed to separate. A pink to red colouration of the classes of antibiotic substances that could serve as
alkaline layer indicated the presence of anthraquinone. selective agents for infectious disease chemotherapy and
control. The effect of these plants on more patho genic
Essential oils: The oil o f A. maritima, G. alata, C. organisms, and toxicological investigations and further
nervatus and C. schoenanthus spp. proximaus was purification, however, need to be carried out.
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