Phytochemistry, Antioxidant and Antibacterial Activities of Medicinal Plants - A Comparative Study
Phytochemistry, Antioxidant and Antibacterial Activities of Medicinal Plants - A Comparative Study
Phytochemistry, Antioxidant and Antibacterial Activities of Medicinal Plants - A Comparative Study
Rajamurugan R. et al
PLANTS- A COMPARATIVE STUDY
ABSTRACT
Plants are the most important sources of medicines. Today the large number of drugs in use is derived
from plants. The important advantages for therapeutic uses of medicinal plants in various ailments are
their safety besides being economical, effective and easy availability. The present investigation was
aimed to screen phytochemicals, antioxidant and antibacterial activities of ethanolic extracts of leaves
of Aegle marmelos, Ocimum sanctum, Alternanthera sessilis, Eclipta alba, and Acalypha indica. The
phytochemical analysis revealed the presence of terpenoids, phenols, flavonoids, tannins, alkaloids,
cardiac glycosides, and steroids in all the extracts. The antioxidant activity of the extracts was
evaluated by DPPH, ABTS and reducing power assay. All the extracts showed significant antioxidant
activity. The plant preparations were also screened individually for antibacterial activity against three
selected bacterial sp by agar-well diffusion method. Results showed that all the extracts were effective
against bacterial sp tested.
Keywords- Aegle marmelos, Ocimum sanctum, Alternanthera sessilis, Antioxidant activity,
Antimicrobial activity.
and leaves are valued in indigenous medicine the treatment of throat infections and wound
(Charakbraty et al., 1960). The plant has been healing, and also used as anti-venom and
employed for long time in folk therapy. Poultice migraine pain relief. (Oudhia, 2003; Valsara,
made of leaves are used for ophthalmia and 1994).
ulcers. The leaves are use to lowering the blood
glucose levels (Ayurvedic Pharmacopoeia of MATERIALS AND METHODS
India, 1988). Other actions like antifungal (Renu, Extraction of plant material
1983), antibacterial (Banerji and Kumar, 1980), The plant materials (Aegle marmelos (AM),
antiprotozoal (Banerjee, 1980), Ocimum sanctum (OS), Alternanthera sessilis
antispermatogenic (Sur et al, 1999) are also (AS) Eclipta alba (EA), and Acalypha indica
reported. Ocimum sanctum Linn. (Labiatae), (AI)) were collected locally. Authentication of
commonly known as holy basil, is an herbaceous the plant materials was made by
plant found throughout the South Asian region Dr.M.Shivashanmugam, Asst professor in
(Hannan et al 2007). The oil of Ocimum Botany, M.G.R College, Hosur. Leaves were
sanctum possesses antibacterial, antifungal, detached and dried in shade. About 100 gms of
antioxidant and radioprotective properties dried leaves were ground to powder and
(Sharma et al., 2002). Ancient Hindu literature is exhaustively extracted with 600 ml ethanol
rich with the medical actions of Ocimum separately using soxhlet apparatus and extracts
sanctum (Ubiad et al., 2002). Alternanthera were concentrated under reduced pressure and
sessilis is a prostrate or perennial herb. The then stored in an air tight container for further
branches are raised from the root and are up to 50 study.
cm long. This plant is found in damp places, wet Phytochemical screening
headlands, roadsides and sometimes as weed of Phytochemical screening was performed using
plantations. In South East Asia this plant is taken standard procedures (Sofowora et al., 1993;
as vegetables. In India it is used for treatment of Trease, 1989).
gastrointestinal problems (Archana et al., 2011). Evaluation of antioxidant activity
Eclipta alba L. Hassk. (Asteraceae) has been DPPH radical scavenging assay
reported for treating liver was cirrhosis and The free radical scavenging activity of all the
infective hepatitis (Chopra et al., 1966). In extracts was measured by decrease in the
Ayurveda, the root powder is used for viz. absorbance of ethanolic solution of DPPH (Braca
treating hepatitis, enlarged spleen and skin et al., 2001). Different concentration of extracts
disorders. Mixed with a little oil when applied to (10-500 µg/ml) and positive control ascorbic acid
the head, the herb relieves headache. The extract was added to 2 ml of freshly prepared DPPH. The
of its leaves is mixed honey and given to infants, measurement was performed in triplicates. After
for to expel worms. Eclipta alba is also given to incubation for about 30 min at room temperature
children in case of urinary tract infections (Prabu in the dark, the absorbance was measured at 520
et al., 2011). Acalypha indica belongs to the nm using spectrophotometer (RAYLEIGH).
family Euphorbiaceae. The leaf can be used for
Radical scavenging activity (%) was calculated using the following formula:-
OD of Control –OD of Sample
DPPH radical scavenging (%) = X 100
OD of Control
antiviral properties (Ahmad et al., 2001; reference compound and that of the ethanolic
Chapagain et al., 2007; Xiong et al., 2011). In extracts. The scavenging effect of AM, OS, AS,
this study, Aegle marmelos, Ocimum sanctum, EA, and AI and standard ascorbic acid on the
Alternanthera sessilis, Eclipta alba, and DPPH radical decreased in the order: ascorbic
Acalypha indica leaves were extracted with acid > AM > AI > OS > EA >AS. The IC50 values
ethanol. The percentage yield was found to be for these compounds were found to be 92 μg/ml,
6.4, 8.8, 5.8, 6.2, and 5.6 respectively (Table 1). 98 μg/ml, 154 μg/ml, 178 μg/ml, 324 μg/ml and
The extracts were screened qualitatively for 522 μg/ml respectively (Table 3). The results also
phytochemicals by chemical method. The results show that the ethanolic extract of AM possesses
of the phytochemical screening revealed the significantly higher DPPH scavenging activity
presence of terpenoids, tannins, steroids, than other extracts and were most potent among
flavonoids, saponins, cardiac glycosides, the five extracts.
alkaloids and phenols in all the extracts. The test ABTS radical scavenging activity
for saponins, however, showed negative result for The ABTS assay is based on the inhibition of the
Eclipta alba, and Alternanthera sessilis (Table radical cation ABTS+, which has a characteristic
2). long wavelength absorption spectrum (Sanchez-
DPPH radical scavenging activity Moreno, 2002). The ABTS chemistry involves
Free radicals have been a subject of significant direct generation of ABTS radical mono cation
interest among scientists in the past decade and with no involvement of any intermediary radical,
their possible role in human diseases has gained it is a decolorization assay, thus the radical cation
importance nowadays (Maxwell, 1995; Jovanovic is formed prior to addition of antioxidant test
and Simic, 2000). Antioxidants neutralize free system rather than the generation of radical
radicals that are defined as atoms or groups of taking place continually in the presence of
atoms having an unpaired electron (Finkel and antioxidant. This method is used for the screening
Holbrook, 2000). These also include related of antioxidant activity of herbal preparations, is
reactive oxygen species (ROS) that leads to free applicable to both lipophilic and hydrophilic
radical generation, causes the cascading chain antioxidants (Long et al., 2000). In this study,
reaction in biological system. Antioxidants ABTS radical was effectively scavenged by the
present in various dietary supplements offered ethanloic extract of AM, followed by AI, OS,
their beneficial effects by neutralizing these ROS EA, and AS in a concentration dependent manner
during various disease conditions. DPPH radical but the scavenging effect was lesser than that of
is widely used as a model to investigate the the reference compound ascorbic acid (Fig.2).
scavenging potential of several natural IC50 values were 102 μg/ ml, 110 μg/ ml, 252 μg/
compounds such as phenolics or crude extract of ml, 288 μg/ ml, and 364 μg/ ml for AM, AI, OS,
plants (Veerapur et al., 2009). In the present EA, and AS respectively. Similarly IC50 for
study, the ethanolic extracts of Aegle marmelos, standard was 96 μg/ ml (Table 4).
Ocimum sanctum, Alternanthera sessilis, Eclipta Reductive ability
alba, and Acalypha indica exhibited marked The reducing capacity of a compound may serve
DPPH radical scavenging activity significantly. as a significant indicator of its potential
Lower absorbance of the reaction mixture antioxidant activity (Blazovics et al, 2003).
indicates higher the free radical scavenging Reducing power is to the measure of the
activity. Fig. 1 shows that % inhibition decreases reductive ability of antioxidant and it is evaluated
the concentration of DPPH radical due to by the transformation of Fe3+ to Fe2+ in the
scavenging ability of standard ascorbic acid, as a presence of sample extracts (Gulcin et al., 2003).
In this study, the reducing capacities of the biological antioxidants (Hagerman, 2002). A
extracts were also determined. Results of the number of mechanisms have been proposed to
reducing power of standard antioxidant ascorbic explain the tannin’s antimicrobial activity which
acid and the ethanolic extracts of AM, OS, AS, includes extracellular microbial enzymes and
EA and AI are shown in Fig. 3. The data show an proteins, deprivation of iron as substances for
increase in the reducing power of the extracts microbial growth or direct action towards its
with increase in dosage. However, the ethanolic membranes (Scalbert, 1991). Condensed tannins
extract of OS presents better activity at a have been determined to bind cell walls of
concentration of 500 µg/ml followed by EA, AS, luminal bacteria, preventing growth and protease
AM, and AI (Table 5). activity (Jones et al., 1994). From the results of
Antibacterial activity this investigation, it is therefore postulated that
Plant based antimicrobial compounds have tannins present in the crude extracts were
enormous therapeutical potential as they can responsible for the antibacterial activity.
serve the purpose without any side effects that are
often associated with synthetic antimicrobials. CONCLUSION
The active ingredients of the plant parts are better From the results of this study it could be
extracted with alcohol than other solvents. The concluded that all the five medicinal plants
alcohol extracts contain alkaloids, coumarins and contain bioactive principles and found to have a
tannins (Okemo, 1996). Coumarins and tannins strong antioxidant and antibacterial activities
have antibacterial and antihelminthic properties specifically in the ethanolic extract of leaves.
(Hedberg et al., 1983). Eloff (1998) and Cowan Results of our findings further confirm the use of
(1999) found that alcohol was more efficient than these herbs as traditional medicine and may be
acetone in extracting phytochemicals from plant used as effective and potential sources of novel
materials. In the present work antibacterial effect antioxidant and antimicrobial drugs.
of leaf extracts from five plants were studied on
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100
Am
0
0 200 400 600
Concentration (g /ml)
Fig. 1. Scavenging activity (%) on DPPH radicals of the extracts. Each value is expressed as mean ±
standard deviation (n = 3).
Am- Aegle marmelos; Os- Ocimum sanctum; As- Alternanthera sessilis; Ea- Eclipta alba; Ai- Acalypha
indica; and ASC- Ascorbic acid
80 Os
As
60 Ea
Ai
40
ASC
20
0
0 200 400 600
Concentration (g /ml)
Fig. 2. Scavenging activity (%) on ABTS radicals of the extracts. Each value is expressed as mean ±
standard deviation (n = 3).
Am- Aegle marmelos, Os- Ocimum sanctum, As- Alternanthera sessilis, Ea- Eclipta alba, and Ai-
Acalypha indica; and ASC- Ascorbic acid
1.5
Am
Absorbance at 700 nm
Os
1.0 As
Ea
Ai
0.5 ASC
0.0
0 200 400 600
Concentration (g /ml)
Fig. 3. Reducing power of the extracts (higher absorbance indicates higher reducing power). Each value is
expressed as mean ± standard deviation (n = 3).
Am- Aegle marmelos, Os- Ocimum sanctum, As- Alternanthera sessilis, Ea- Eclipta alba, and Ai-
Acalypha indica; and ASC- Ascorbic acid
30
20
Am
Os
10 As
Ea
Ai
0
25 100 250 500
Concentration (g /ml)
Fig.4. Antibacterial activities of the extracts against E. coli (agar-well diffusion method). Am- Aegle
marmelos, Os- Ocimum sanctum, As- Alternanthera sessilis, Ea- Eclipta alba, and Ai- Acalypha indica
30
20
Am
Os
10 As
Ea
Ai
0
25 100 250 500
Concentration (g /ml)
Fig.5. Antibacterial activities of the extracts against S. aureus (agar-well diffusion method). Am- Aegle
marmelos, Os- Ocimum sanctum, As- Alternanthera sessilis, Ea- Eclipta alba, and Ai- Acalypha indica
20
Am
Os
10 As
Ea
Ai
0
25 100 250 500
Concentration (g /ml)
Fig.6. Antibacterial activities of the extracts against Pr. mirabilis (agar-well diffusion method). Am-
Aegle marmelos, Os- Ocimum sanctum, As- Alternanthera sessilis, Ea- Eclipta alba, and Ai- Acalypha
indica.
Fig.7. Antibacterial activities of the extracts (agar-well diffusion method). 7 A- Aegle marmelos,
S.aureus; 7 B- Aegle marmelos Pr. mirabilis; 7 C- Aegle marmelos, E. coli; 7 D- Ocimum sanctum,
S.aureus; 7 E- Ocimum sanctum, Pr. mirabilis; 7 F- Ocimum sanctum, E. coli; 7 G- Alternanthera
sessilis, S.aureus; 7 H- Alternanthera sessilis, Pr. mirabilis; 7 I- Alternanthera sessilis, E. coli; 7 J-
Eclipta alba, S.aureus; 7 K- Eclipta alba, Pr. mirabilis; 7 L- Eclipta alba, E. coli; 7 M- Acalypha indica,
S.aureus; and 7 N- Acalypha indica, Pr. mirabilis;7 O- Acalypha indica, E. coli.