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Evaluation of Anti-Microbial, Anti-Inflammatory and Anti-Oxidative

Properties Artemisia afra, Gunnera perpensa and Eucomis autumnalis

Article · September 2014

DOI: 10.4172/2155-9600.1000312

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 Nutrition and Food Muleya et al., J Nutr Food Sci 2014, 4:6
http://dx.doi.org/10.4172/2155-9600.1000312
Sciences

Research Article Open Access

Evaluation of Anti-Microbial, Anti-Inflammatory and Anti-Oxidative

Properties Artemisia afra, Gunnera perpensa and Eucomis autumnalis
Muleya E1*, Ahmed AS2, Sipamla AM1, Mtunzi FM1 and Mutatu W3
1
Department of Chemistry, University of Technology, P. Bag 021, Vanderbijlpark, 1900 South Africa
2
Phytomedcine Programme, University of Pretoria, P. Bag X04, 0110 Onderstepoort, South Africa
3
Midlands State University, Department of Chemical Technology, P. Bag 9055, Gweru, Zimbabwe

Abstract
Antimicrobial, anti-inflammatory and free radical scavenging activities of crude root extract fractions from
Artemisia afra, Gunnera perpensa and Eucomis autumnalis were determined. Minimum inhibitory concentration was
determined by using micro dilution method. In order to assess antioxidant scavenging capacity of plant extracts and
fractions, 2, 2-di (4-tert-octylphenyl)-1-picrylhydrazyl and 2, 2´-azinobis (3-ethylbenzothiazoline)-6-sulfonic was used
as substrate. Anti-inflammatory activity of the plant extracts against 15-soybean lipoxygenase enzyme was evaluated
by measuring change in absorbance at 234 nm using linoleic acid as substrate. The highest activity was obtained
from methanol fraction of Gunnera perpensa with EC50 value of 1.069 µg/ml against 2,2-di (4-tert-octylphenyl)-1-
picrylhydrazyl. Eucomis autumnalis crude and acetone fraction displayed DPPH• free radical scavenging activity
of EC50 of 2.891 µg/ ml and 2.41 µg/ ml respectively. Artemisia afra crude fraction and fractions of acetone and
methanol displayed activity (EC50 for DPPH• radical, 2.113 µg/ ml with crude 4.393 µg/ ml with acetone fraction, 4.715
µg/ ml, with methanol fraction and with ABTS•+ radical cation, 6.447 µg/ ml and 6.208 µg/ ml from crude and methanol
fraction respectively). The antioxidant properties of the extracts increased with the polarity of the fractions. Gunnera
perpensa crude extract and fractions displayed antimicrobial properties with the methanol fraction being the most
active with an EC50 of 80 µg/ ml, against Pseudomonas aeruginosa and EC50 of 160 µg/ml against Candida albicans.
Artemisia afra acetone and methanol fractions displayed inhibitory activities of 20 µg/ml against Escherichia coli and
good-moderate activity ranging 160-320 µg/ml for the crude extract. Eucomis autumnalis had activities ranging 160-
320 µg/ml by the crude extract and fractions against the organisms tested except for crude extract activity against
Escherichia coli of 630 µg/ml. The activities validate claims by the traditional healers use for cure offering possible
alternative as dietary supplements to the management of inflammation related conditions.

Keywords: Minimum inhibitory concentrations (MIC); Reactive [8].The beneficial medicinal effects of plant phytochemicals typically
oxygen species (ROS); Reactive nitrogen species (RNS). result from the synergism of secondary products present in the plant
[9].
Abbreviations: ABTS•+: 2,2´-Azinobis(3-ethylBenzothiazoline)-
6-Sulfonic acid radical; DPPH•: 2,2-Di(4-tert-octylPhenyl)-1- Infection is usually accompanied by microbial invasion followed
PicrylHydrazyl radical; 15-LOX: 15-Soybean Lipoxygenase by the occurrence of oxidative stress and serious inflammation
[7]. Activation and proliferation of pro-inflammatory cytokines in
Introduction respiratory epithelial cells and macrophages are down regulated
by supplying and maintaining sufficient levels of exogenous and
Infectious disease is an illness resulting from the invasion of the
endogenous antioxidants [10].
host species by a pathogenic microbial agent, and outcome of the
disease depends on the degree of success of the invading pathogen and Phytochemicals from medicinal plants have provided unlimited
immune system of the host [1]. They are considered a major threat opportunities for new drug discoveries because of their inherent
to human health, because of the unavailability of vaccines or limited chemical diversity, which has prompted a continuous search for plant
chemotherapy even in the developed parts of the world, although sources with medicinal value [11,12]. Artemisia afra Jacq. ExWilld.
developing countries are carrying the major part of the burden Sub (Asteraceae), Gunnera perpensa L. (Gunneraceae) and Eucomis
Saharan African countries, including South Africa are mostly affected autumnalis (Mill) (Hyacinthaceae) Chitt are medicinal plants mainly
by respiratory infections, diarrhea, HIV/AIDS, tuberculosis and malaria used in the Mabandla village of UMzimkhulu Local Municipality,
[2,3] The continued resurfacing of antibiotic- resistant infections
drives research to produce better drugs to combat the more resistant
pathogens [4]. Auto immune diseases such as lupus are becoming a
*Corresponding author: Muleya E, Midlands State University, Department of
major concern in both the developed and third world countries [5]. Chemical Technology, P. Bag 9055 Gweru. Zimbabwe, Tel: +263 778 739 201;
Investigating plants that could be included in affected people’s diet E-mail: e_muleya@yahoo.co.u, tarirogopoza@gmail.com
would assist in managing such diseases.
Received July 07, 2014; Accepted September 22, 2014; Published September
Fighting infections with natural products will be more 27, 2014
advantageous and affordable in the southern African context over the Citation: Muleya E, Ahmed AS, Sipamla AM, Mtunzi FM, Mutatu W (2014)
conventional drugs to most patients since they are readily available in Evaluation of Anti-Microbial, Anti-Inflammatory and Anti-Oxidative Properties
Artemisia afra, Gunnera perpensa and Eucomis autumnalis. J Nutr Food Sci 4:
the environment in which they live and some are cultivated to ensure 312. doi: 10.4172/2155-9600.1000312
product quality and safety [6]. Antimicrobials of plant origin have
enormous therapeutic potential in the treatment of infectious diseases Copyright: © 2014 Muleya E, et al. This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which permits unrestricted
[7]. They are effective as well as have the advantage of mitigating many use, distribution, and reproduction in any medium, provided the original author and
of the side effects that are often associated with synthetic antimicrobials source are credited.

J Nutr Food Sci

ISSN: 2155-9600 JNFS, an open access journal Volume 4 • Issue 6 • 1000312
Citation: Muleya E, Ahmed AS, Sipamla AM, Mtunzi FM, Mutatu W (2014) Evaluation of Anti-Microbial, Anti-Inflammatory and Anti-Oxidative Properties
Artemisia afra, Gunnera perpensa and Eucomis autumnalis. J Nutr Food Sci 4: 312. doi: 10.4172/2155-9600.1000312

Page 2 of 6

Kwa-Zulu Natal, South Africa for the treatment of infectious and extract was concentrated. The concentrated slurry was dried at room
some inflammatory mediated diseases (Table 1). Therefore, it is of temperature in the fume hood. The crude plant extract was stored in
interest to observe if these plant extracts can affect these mechanisms the fridge at 4°C. Fractions of different polarities were then produced
of pathogenic changes that lead to diseases, if over-expressed. This by solvent/ solvent extraction from the crude extracts as shown in the
study focuses on the antimicrobial, antioxidant and anti-inflammatory protocol in Figure 1. Percentage yields for the crude, and the different
properties of selected plant extracts, namely their ability to directly fractions were reported in Table 2.
scavenge free radicals; their effects on soya bean derived 15 LOX
inhibitory activities and inhibition of microbial growth (Staphylococcus Quantitative evaluation of the biological activities of the
aureus, Enterococcus faecalis, Escherichia Coli, Pseudomonas aeruginosa plant extracts
Aspergillus fumigatus, and Candida albicans). Determination of antimicrobial activity: The following
microorganisms were used as test organisms in the screening: two Gram-
Material and Methods
positive reference strains (Staphylococcus aureus (ATCC 29213) and
Plant selection and collection Enterococcus faecalis (ATCC 29212)), and two Gram-negative reference
strains (Pseudomonas aeruginosa (ATCC 27853), and Escherichia coli
Artemisia afra, Gunnera perpensa and Eucomis autumnalis (Table 1)
from Mabandla village of UMzimkhulu Local Municipality, KwaZulu-
Natal, and South Africa were used for the study. They were identified
and collected by the aid of the head of traditional healers from the Plant dry powder

village (Mr. Sanoyi Paulos Dlamini, Traditional healer, KwaZulu-

Acetone extract
Natal 2009). Authentication of plants was carried out at South Africa Drying using rotary
National Biodiversity Institute, Pretoria and their voucher specimens Dried extract
evaporator

were deposited in the Pretoria National Herbarium.

Plant treatment Dissolve in 70% acetone Fractionation with solvent of
different polarities

Plant root and bulb materials were washed in water, air dried at
room temperature for three weeks and ground to powder using a Lasec Hexane fraction Dichloromethane Ethyl acetate Residual water
Polymix PX-MFC 90D crusher. The dried pulverized material was fraction fraction fraction

stored in glass containers in a cool dry place. The extraction was carried
Acetone soluble fraction Methanol soluble fraction
out by shaking the powder (200 g) in a ratio of 1 g to 10 ml solvent for
6 h on a Remi R S-12R shaker (Rajendra Electrical India Ltd, India) at Figure 1: Extraction and fractionation procedures.
200 rpm [13]. Excess solvent was recovered on a rotary vapor until the

Botanical name and

Biological activities and isolated compounds from previous
specimen Voucher South African name Traditional medicinal use
studies
numbers
Umhhlonyane (Xhosa),
Among the volatile secondary metabolites, there are 83
Mhlonyane (IsiZulu), Lanyana Rhizomes and roots are used for,
monoterpenoids, 8and 50 sesquiterpenes and under the non-volatile, 2
Artemisia afra: 3166- (Sesotho), Lengana (Tswana), colds, influenza, stomach and
sesquiterpens, 7 glaucolides,6 guaianolides,.4 triterpenes, 6 long chain
4001 African wormwood (English), respiratory ailments, rheumatism and
alkanes,5 coumarins,1 organic acid,1 glycoside,11 flavonoids (Watt and
and Wilde also (Afrikaans). Van wounds(Somova et al (2001)
Breyer- Branwijk, (1962); Van Staden et al., 2009) Liu et al, 2008).
Staden et al., (2009)
Roots extracts are used to aid
2-methyl-6-(3-methyl-2-butenyl) benzo-1,4-quinone and - hydrxxy-
pot-partum expulsion, relief of
Gunnera perpensa: 8-methyl-2,2- dimethyl-2H-benzopyran from Gunnera perpensa.
Ugobho (isiZulu) dysmenorrhea, psoriasis and cure for
2537-1 Respective MIC range = 70 µg/ml and 75 µg/ml of isolate (Drewes et
female infertility (Kaidoo, T. L., (1997)
al., 2005)
Van Wyk et al, (1997))
3,7-dihydroxy-1methyl-6H-dibenzo[b,d,]pyran-6-one(autumnariol)
3,4,7-trihydroxy-1-methyl-6H-dibenzo[b,d]pyran-6-one (autumnariol)
5,7,dihydroy-8methoy-3-(4’-methoxybenzyl)-4-chromanone(4’-O-
methyl-3,9-dihydropunctatin)
5,7-dihydroxy-6-methoxy-3-(4’-hydrxybenzyl-4-chromanone(3,9-
dihydrocucomnalin or 3,9-dhydroautumnalin)
3,5,7-trihydroxy-3-(4’-methoxybenzyl)-4-chromanone(eucomol)
(E)-5,7-dihydroy-6-methoxy-3-(4’-hydroxybenzylidene)-4-
chromanone(autumnalin) Bulbs are used for a variety of
(Z)-5,7-dihydroxy-6-methoxy-3-(4’-hydroxybenzylidene)-4- ailments including urinary diseases,
Eucomis autumnalis: Umathunga (isiZulu) Khapumpu
chromanone(autumnalin) stomach ache and fevers. (Jager et
3790-4001 (Sesotho)
5,7-dihydroxy -8-methoxy-3-(4’-methoxybenzylidene)4-chromanone(4’- al, (1996); Taylor and van Staden,
O-methylpunctatin) (2001); Stafford et al., (2004).
(23S)-17,23-epoxy-3β,29-dihydroxy-27-norlanost-8-ene-15,24-
dione(eucosterol)
(23S)-17,23-epoxy-3β,16β,29-trihydroxy-27-norlanost-8-ene-15,24-
dione(16β-hydroxyeucosterol)
3β-hydroxy-4β-hydroximethyl-4α,14α-dimethy-15-oxo-24-nor-5α-chola-
8,16-dien-23-oic acid
R-(-)-2-(4’-hydroxybenzyl)-malic acid (eucomic acid) (Taylor and van
Staden,(2001); Stafford et al., (2004,)
Table 1:Ethno pharmacological information of the plant use in this study.

J Nutr Food Sci

ISSN: 2155-9600 JNFS, an open access journal Volume 4 • Issue 6 • 1000312
Citation: Muleya E, Ahmed AS, Sipamla AM, Mtunzi FM, Mutatu W (2014) Evaluation of Anti-Microbial, Anti-Inflammatory and Anti-Oxidative Properties
Artemisia afra, Gunnera perpensa and Eucomis autumnalis. J Nutr Food Sci 4: 312. doi: 10.4172/2155-9600.1000312

Page 3 of 6

Plant species C H DCM ET AC Met W a blank solution of 40 μl solvent and 160 μl ABTS•+ . For the positive
control, 160 μl solution of ABTS•+ in methanol was mixed with 40 μl
Artemisia afra 2.73 0.55 12.08 1.09 57.69 1.09
methanol. Trolox and ascorbic acid were used as positive control. The
Gunnera perpensa 6.45 1.70 0.39 0.46 4.03 40.23 38.20
assay was carried out in triplicate wells on each plate and with three
Eucomis autumnalis 2.66 0.19 4.71 0.56 0.56 56.12
independent experiments. Percentage inhibition of free radical ABTS•+
C= crude extract, H= hexane fraction, DCM= dichloromethane fraction, ET = ethyl was calculated thus: Percentage inhibition (%IP) = [(A t=0 min – A t=30
acetate fraction, AC = acetone fraction and Met= methanol fraction, W= water
fraction min
)/At=0 min]×100 The EC50 value (μg/ml) (concentration of extract
Table 2: The % yield of the crude and fractions of the extracts. required to inhibit ABTS•+ radical formation by 50%) was determined
via extrapolation from best fit non-linear analysis, using the Graph Pad
(ATCC 25922)). In addition, two pathogenic clinical isolate of fungi Prism software (California, USA).
(Candida albicans and Cryptococcus neoformans (supplied by the
Phytomedicine laboratory, University of Pretoria, South Africa) were In vitro lipoxygenase inhibition assay: The soybean derived
also employed as test organisms. Stock cultures were maintained at 4°C 15- lipoxygenase type I-B (15-LOX) was evaluated as described by
on slopes of nutrient agar. The cultures were diluted to achieve optical Malterud and Rydland (2000) [16]. The reaction mixture contained 20
densities corresponding to 2.0×106 colony forming units (CFU/ml) for mM borate HCl buffer (pH 8.8), test compound, linoleic acid solution
bacterial and 2.0×105 spore/ml for fungal strains. (40 μM), and soybean lipoxygenase (131.000 U/mL). For inhibition
experiments, 15-LOX was incubated with inhibitor for 5 min in a 10
Antimicrobial assay mm path length cuvette at room temperature. The reaction was then
Inoculate of the bacteria were made from 24 h Mueller–Hinton started by the addition of 10 μl of linoleic acid solution (substrate). The
broth (Sigma) cultures and standardized using 0.5 McFarland standards control was run in DMSO the solvent used to dissolve the inhibitor.
comparable to a bacterial suspension of 1.0×108 cell/ml. Crude extracts After 5 min, the absorbance was read at 234 nm as a measure of the
and fractions of various polarities were reconstituted in 70% acetone conjugated diene produced. For the determination of IC50, the inhibitor
(10 mg/ml). Gentamicin and 70% acetone were used as the positive was varied at the constant substrate concentration of 80 μM linoleic
and negative controls respectively. Minimum inhibitory concentration acid. The % inhibition of 15-LOX activity by extract was calculated
(MIC) values of the samples and controls were determined using a micro- from the absorbance values at 234 nm at the end of 5 min: Inhibition
well dilution method over three independent experiments [13]. The (%) = (Ac – As/Ac)×100 where Ac and As were absorbance of the control
sterile 96-well microtitre plates were prepared with 100 μl of Millipore (without test sample) and the sample, respectively. All measurements
filtrated water per well and a two-fold serial dilution was prepared from are an average of triplicate measurements and expressed as the 50%
100 μl crude extracts or fractions in 70% acetone (10 mg/ml) added inhibition concentration value (IC50) from the control without inhibitor.
to the first well. An aliquot (100 μl) of the standardized inoculum was The enzyme solution was kept on ice, and controls were measured
added to wells and the plates were incubated overnight at 37°C. The at intervals throughout the experimental periods to ensure that the
final concentration in wells ranged from 2.5 to 0.019 mg/ml. Bacterial enzyme activity was constant.
growth was determined by adding 40 μl p-iodonitrotetrazolium (INT)
Results and Discussion
violet (Sigma, UK) 2 mg/ml in water. The extracts were then incubated
for 30 min and bacterial growth was determined by INT formazan The antioxidant scavenging capacity of plant extracts and fractions
production. were determined using the DPPH• and ABTS• free radical scavenging
assays and the results were expressed as EC50 as presented in Table 2 and
DPPH radical scavenging assay: The DPPH scavenging activity of
antimicrobial assays in Figures 2-4. The results for the 15-LOX activities
the extracts was measured from the bleaching of the of purple DPPH are summarized in Table 3. The percentage yield (Table 2) of fractions
solution in methanol [14]. Twenty microliter extract or reference showed that for Gunnera perpensa and Eucomis autumnalis more than
standards in methanol (500-1.9 μg/ml) were added to 180 μl of 6.5×10−5 50% of total extract for each plant was in the polar solvents of methanol
M DPPH methanol solution in 96-well microtitre plates. The plates were and water.
incubated at room temperature in the dark for 30 min after which the
optical density was recorded at 517nm using a microtitre plate reader.
For the positive control, 160 μl solution of DPPH in methanol was mixed
with 40 μl methanol. Trolox and ascorbic acid were used as positive
control. The assay was carried out in triplicate wells on each plate and
with three independent experiments for 30 min. Percentage inhibition
of free radical DPPH was calculated thus: Percentage inhibition (%IP) =
[(A t=0 min – A t=30 min)/At=0]×100 . The EC50 value (μg/ml) (concentration
of extract required to inhibit DPPH radical formation by 50%) was
determined via extrapolation from best fit non-linear analysis, using
the Graph Pad Prism software (California, USA).
ABTS•+ scavenging assay: The scavenging activity against
ABTS cation radical was assayed to determine the Trolox Equivalent
Antioxidant Capacity value (TEAC) using a reported method [15] The
blue-green ABTS•+ was produced by reacting 7 mM ABTS and 2.45
mM potassium persulfate in water and the solution was diluted with Figure 2: Antimicrobial activities of Eucomis autumnalis crude extracts and
methanol to a final absorbance of 0.7 ± 0.02 at 734 nm. Forty microliters fractions of different polarities as indicated by solvent used: S.a= Staphylococcus
aureus, Ef = Enterococcus faecalis, E.c. = Escherichia coli, P.a. = Pseudomonas
of antioxidants solution were added to 160 μl ABTS•+ solution, mixed
aeruginosa, C.a. = Candida albicans, A.f= Aspergillus fumigatus.
vigorously, and measured promptly at the absorbance 734 nm against

J Nutr Food Sci

ISSN: 2155-9600 JNFS, an open access journal Volume 4 • Issue 6 • 1000312
Citation: Muleya E, Ahmed AS, Sipamla AM, Mtunzi FM, Mutatu W (2014) Evaluation of Anti-Microbial, Anti-Inflammatory and Anti-Oxidative Properties
Artemisia afra, Gunnera perpensa and Eucomis autumnalis. J Nutr Food Sci 4: 312. doi: 10.4172/2155-9600.1000312

Page 4 of 6

Antimicrobial assays dilution method. Ethno medicine is one method of discovering new
drugs which may offer a solution to cure for disease caused by pathogens
Gunnera perpensa crude extract and fractions displayed high which have resistance to known drugs [17]. In this investigation, Gunnera
activities (Figure 4) against the organisms tested using the micro plate perpensa extracts and fractions displayed good broad based antimicrobial
activities against the organisms tested using the micro plate dilution
method. The most active fraction was the methanol fraction (80 µg/ ml)
against Pseudomonas aeruginosa. Gunnera perpensa fractions (hexane,
DCM, ethyl acetate, acetone and methanol) have good activity of 160 µg/
ml against Candida albicans. The antimicrobial activities of water, ethyl
acetate and ethanol fractions of the root extract of Gunnera perpensa
against some bacteria and fungi has been reported before for Bacillus
subtilis, 12.5 mg/ml by Buwa and Van Staden [18]. Crude water extract
of Gunnera perpensa have antimicrobial activity against Escherichia
coli (0.78 mg/ml); Klebsiella pneumoniae (0.78 mg/ml); and Candida
albicans (25 mg/ml) while the corresponding ethyl acetate and ethanol
fractions were less active (Buwa and van Staden. However, the reported
values are not considered to be significantly high by the phytomedicine
program. This explains the use of the plant by South African Traditional
Figure 3: Antimicrobial activities of Artemisia afra crude extracts and fractions healers as treatment against venereal diseases [19]. Drewes et al., isolated
of different polarities as indicated by solvent used: S.a= Staphylococcus aureus,
E.f= Enterococcus faecalis, E.c. = Escherichia coli, P.a. = Pseudomonas 1,4 –benzoquinone derivatives, which were identified as 2, methyl-6-
aeruginosa, C.a. = Candida albicans, A.f= Aspergillus fumigatus. (3-methyl-2-butenyl) benzo-1,4- quinone (MIC of 18 µg / ml against
Bacillus cereus) and 3-hydroxy-2-5-(3-methyl-2-butenyl)benzo-1,4-
quione (MIC of 37 µg / ml against Candida albicans and 75 µg / ml and
against Cryptococcus neoformans) the active antimicrobial components
against the bacteria and fungus [19]. Although this characterization is
not exhaustive, it explains why Gunnera perpensa is so active against the
organisms tested (Table 4). South African traditional healers use Gunnera
perpensa aqueous decoction to inducing labor, facilitating the expulsion
of placenta and relief of dysmenorrhea [20]. Gunnera perpensa also
displayed antinociceptive and anti-inflammatory activity [21].
Artemisia afra acetone and methanol fractions have good inhibitory
activities (20 µg/ml) against E. coli and good-moderate activity
ranging 160-320 µg/ml for the crude extract and all fractions against
the organisms tested (Figure 3) against Aspergillus fumigatus and
Staphylococcus aureus. Artemisia afra is used in traditional medicine
in the treatment of a variety of diseases ranging from respiratory
Figure 4: Antimicrobial activities of Gunnera perpensa crude extracts and infections to dysmenorrhea, diabetes and malaria [22,23]. The plant
fractions of different polarities as indicated by solvent used: S.a= Staphylococcus rich in terpenes such as artemisia alcohol, camphene, camphor and
aureus, E.f= Enterococcus faecalis, E.c. = Escherichia coli, P.a. = Pseudomonas artemisia ketone [23] and displayed a variety of biological activities
aeruginosa, C.a. = Candida albicans, A.f= Aspergillus fumigatus.
(using disc diffusion method) against bacteria such as Staphylococcus
aureus (MIC 2.0 mg/ml), Mycobacterium smegmatis (MIC 1.9 mg/ ml)
plant species EC50 fungi such as Candida albicans (% minimum inhibitory percentage of
G. Perpensa 81.18 0.25) and protozoa such as P falciparum (IC50 of 4,4 µg/ml) tested in the
E. autumnalis 42.76 investigations [23]. This explains why the percentage yield for the polar
A. afra 21.84
fraction from Artemisia afra (Table 2) was lower compared to the other
two plants. Artemisia afra is used to treat various inflammatory related
Table 3: Soya bean based 15 LOX inhibitory activity expressed as EC50 values for
the crude in 25 µg/ml.
diseases as displayed in Table 4.

Sample A. afra G. perpensa E. autumnalis

DPPH ABTS DPPH ABTS DPPH ABTS
EC50 R2 EC50 R2 EC50 R2 EC50 R2 EC50 R2 EC50 R2
Cr 2.113 0.5043 6.447 0.9697 1.069 0.346 32.49 0.9302 2.891 0.7544 12.18 0.8771
H 29.5 0.4448 96.66 0.8541 46.88 0.5588 646.5 0.4257 21.67 0.5432 929.40 0.6797
DCM 36.03 0.8452 89.08 0.9166 48.71 0.8813 164.9 0.6992 87.71 0.9615 130.00 0.726
ET 199.3 0.6795 68.57 0.9308 57.67 0.7889 11.39 0.8678 95.32 0.8442 24.44 0.9709
Ac 4.393 0.9288 32.95 0.8637 2.795 0.2938 261.5 0.4348 2.461 0.7564 24.44 0.9709
Met 4.715 0.8696 6.028 0.8994 2.795 0.2938 292.9 0.4577 6.621 0.7286 15.17 0.9446
Cr=crude extract, H=hexane fraction, DCM=dichloromethane fraction, ET=ethyl acetate fraction, Ac=acetone fraction, Met=methanol fraction, Positive standards: trolox
=0.04 ± 0.001, ascorbic acid=0.04 ± 0.001
Table 4: Free radical scavenging activity of the crude extract and fractions of varied polarities (μg/ml).

J Nutr Food Sci

ISSN: 2155-9600 JNFS, an open access journal Volume 4 • Issue 6 • 1000312
Citation: Muleya E, Ahmed AS, Sipamla AM, Mtunzi FM, Mutatu W (2014) Evaluation of Anti-Microbial, Anti-Inflammatory and Anti-Oxidative Properties
Artemisia afra, Gunnera perpensa and Eucomis autumnalis. J Nutr Food Sci 4: 312. doi: 10.4172/2155-9600.1000312

Page 5 of 6

Eucomis autumnalis displayed good to moderate (160-320 µg/ml) indomethacin (0.5 μ/ml) at 65% and also displayed selective COX-
activity from the crude extract and fractions against the organisms tested 2 activity [26,27]. Gunnera perpensa acetone, methanol fraction and
except for the crude extract against E coli in which the activity of 630 µg/ crude extract displayed high activity of DPPH free radical scavenging
ml, which was low. The various antimicrobial activities demonstrate the of 2.795 µg/ ml (Table 4) illustrating ant oxidant capability. In previous
validity of the healing capacities of the plant that the traditional healers research, Gunnera perpensa methanol extracts demonstrated strong
of the community claim it possesses. Invasion of the body by organisms ABTS•+ (78.45) and DPPH• (78%) scavenging at a concentration of
such as Escherichia coli, Candida albicans, Pseudomonas aeruginosa and 50 µg/ ml [21]. Gunnera perpensa EC50=81.18 µg/ ml exhibited some
other such pathogens can result in chronic inflammation [7]. soya bean 15-LOX inhibitory activity. Gunnera perpensa methanol
and aqueous extracts has been demonstrated to possess analgesic and
Antioxidant and 15-LOX activity anti-inflammatory activity before [21]. In previous research, Eucomis
The highest activity is obtained from methanol fraction of Gunnera autumnalis also inhibited totally prostaglandin synthesis and displayed
perpensa with EC50 value of 1.069 mg/ml against DPPH•, EC50. preferential COX-2 inhibitory activity [6] and these pharmacological
The lowest activity was displayed by hexane fraction from Eucomis properties make the plant suitable for treating inflammation and would
autumnalis (929.4 µg/ ml). Gunnera perpensa crude and fractions were be ideal as dietary supplements to sufferers of autoimmune diseases and
more active with DPPH• (57.67 mg/L) than with ABTS•+ (11.39 µg/ ml) other inflammation related conditions.
except for the ethyl acetate fraction. Eucomis autumnalis crude and
acetone fraction displayed high DPPH• free radical scavenging activity Conclusions
of EC50 of 2.891 µg/ ml and 2.41 µg/ ml respectively. ABTS•+ free radical The plants in this investigation exhibited good anti-microbial
activity for the plant sample was generally lower for all samples except activity on the organisms tested. Gunnera perpensa and Eucomis
for the ethyl acetate moderate activity of 24.44 µg/ ml. Artemisia afra autumnalis methanol fractions exhibited high free radical activity
crude extract and methanol fraction DPPH• and ABTS•+ free radical against both ABTS•+ and DPPH• radicals displaying some nutritional
scavenging activity as displayed by the curves on Figures 2 to 4 and value. Biological activities of fractions of different polarities (from
the EC50 values. High activity is also displayed by the acetone fraction hexane to methanol) were highlighted in this study are compared to
DPPH• anti-radical activity Gunnera perpensa had the highest amount reports that have been made before of the same plants but different
of yield of extract of 6.2% and 38% of the Gunnera perpensa extract was fractions from previous studies. These plants are used for treating
water soluble. Acetone and methanol extracts were also much higher the various disorders related to inflammation by the traditional
for Gunnera perpensa and Eucomis autumnalis extracts. ’The minimum Zulu practitioners (Table 1). The activities also validate claims by the
inhibitory concentrations of the methanol crude extracts and fractions traditional healers’ use of plants for cure. These results were produced
were presented in Table 4. from in vitro assays using artificial radicals ABTS, DPPH, microbes,
Artemisia afra forms part of southern African indigenous medicines Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis,
with a range of applications including treatment of conditions associated Escherichia coli, Candida albicans and Aspergillus fumigatus therefore
with chronic inflammation such as rheumatism fever, diabetes, asthma, the plant extracts and fractions activity in vivo still need to be carried
malaria and wounds [23,24] A. afra volatile oils have been reported out in order to fully validate their activities as antioxidants in the body.
to demonstrate antioxidant activity when sprayed by DPPH• during a The soya bean derived 15-LOX enzyme inhibitory activities were used
TLC screening method of evaluation and during non-enzymatic lipid only to serve as a guideline to demonstrate inhibitory ability. In vivo
peroxidation in liposomes [24]. In this investigation, high anti-oxidant experiments using human derived 15-LOX still need to be carried
activities were displayed (Table 5). Simelane et al. 2010 have reported out to provide a better picture of the inhibitory activities of the crude
DPPH and ABTS scavenging activity IC50 of 1.6 µg/ ml for DPPH and extracts reported here.
0. µg/ ml for ABTS values which are comparable (EC50 of 1-2.8 µg/ ml) Acknowledgements
to those obtained in this study.
University of Pretoria Phytomedicines Programme, Vaal University
Several reports on the biological activities of Artemisia afra have of Technology for funding the study, Traditional healers of Mabandla
been made before including the isolation and characterization of Village, KwaZulu-Natal, South Africa, Midlands State University,
volatile and nonvolatile metabolites [25]. The fact that Artemisia afra Gweru Zimbabwe
crude fraction and fractions of acetone and methanol displayed high
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J Nutr Food Sci

ISSN: 2155-9600 JNFS, an open access journal Volume 4 • Issue 6 • 1000312
Citation: Muleya E, Ahmed AS, Sipamla AM, Mtunzi FM, Mutatu W (2014) Evaluation of Anti-Microbial, Anti-Inflammatory and Anti-Oxidative Properties
Artemisia afra, Gunnera perpensa and Eucomis autumnalis. J Nutr Food Sci 4: 312. doi: 10.4172/2155-9600.1000312

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