Studies on Phytochemical Screening, Antimicrobial and Anti Radical Scavenging
Effect Coastal Salt Mash Plant of a Suaeda monoica
Salt mash plants have used in many country for food and feed. The coastal people
utilized the folk medicine based on the traditional knowledge to many diseased.
Generally salt tolerance plants have more phyto chemical and more anti oxidant
effect indicated that lot of literature, So in this way in our present study
antioxidant, antimicrobial and phytochemcial screening of coastal salt mash
plant of a Suaeda monoica. To evaluate the medicinal value of Suaeda
monoica and we choose tree types of extracts, (Petroleum ether, ethyl acetate
and methanol), three types of concentration (100, 50, 25 mg mL-1)
and three types of activity (photochemical screening, anti microbial and antioxidant).
In our result point out maximum anti microbial effect observed in petroleum
ether extract at 100 mg mL-1 concentration and minimum anti microbial
effect observed in menthol at 25 mg mL-1 concentration. In the case
of phytochemical screening was observed in phenolic group for all the extract
and alkaloids, anthraquinones, catechins, flavonoids, gum, oils and resins and
saponins are completely absent in all extracts.. Antioxidant effect maximum
(71.4) in methonal extrat at 100 mg mL-1 concentration and minimum
in 28.5 for at 25 mg mL-1 concentration of petroleum ether extract.
to cite this article:
K. Muthazhagan, P. Thirunavukkarasu, T. Ramanathan and D. Kannan, 2014. Studies on Phytochemical Screening, Antimicrobial and Anti Radical Scavenging
Effect Coastal Salt Mash Plant of a Suaeda monoica. Research Journal of Phytochemistry, 8: 102-111.
Received: December 24, 2013;
Accepted: March 02, 2014;
Published: May 05, 2014
Suaeda monoica Forsk. ex. Gmel (Chenopodiacea) is a salt marsh mangrove
herb similar to Suaeda maritima L. Dumort in appearance, growing in hypersaline
soils. It is distributed throughout the East West coast mangroves in India viz.,
Sunderbans in West Bengal State, Bitharkanika and Mahnadhi in Orissa State,
Coringa, Godavari and Krishna in Andhra Pradesh State, Pichavaram, Karangadu
and Muthupet in Tamil Nadu State. It is a shrub but much smaller in size (0.3-0.7
mm in length) when compared to Suaeda maritima. Leaves simple, succulent,
linear, young twigs are slender ribbed. Locally it is called as Vellaikirai
(or) Nilavumari (seaside Indian salt wort). The leaves have been used as edible
green leaves. The ash obtained from burnt plant parts have been exported without
knowing the purpose. It is a folklore medicinal plant used mainly against rheumatism,
paralysis, asthma and snake-bites, skin disease, ulcer for local coastal people
(Kathiresan and Ramanathan, 1997; Ramanathan,
2000). Traditionally, the leaf from Suaeda monoica is known to use
as a medicine for hepatitis (Bandaranayake, 1998) and
scientifically it is reported to be used as ointment for wounds (Padmakumar
and Ayyakkannu, 1992) and possess antiviral activity (Premnathan
et al., 1992) because of the presence of triterpenoids, sterols (Ghosh
et al., 1985; Subramanyam et al., 1992).
The hepatoprotective evaluation of crude ethanolic extract from leaves of S.
monoica for possible development of hepatoprotective herbal medicine (Ravikumar
et al., 2010). Anti-oxidants are vital substances which possess the
ability to protect the body from damage caused by free radical induced oxidative
stress (Ozsoy et al., 2008). There is an increasing
interest in natural anti-oxidants, e.g., polyphenols, present in medicinal and
dietary plants which might help prevent oxidative damage (Silva
et al., 2005). Polyphenols possess ideal structural chemistry for
free radical scavenging activity and they have been shown to be more effective
anti-oxidants in vitro than tocopherols and ascorbate. Antimicrobial
activity of different parts of this plant which has not been reported; hence,
the present study was undertaken. The phytochemical literature reveals the presence
of 2-benzoxazolinone, lignan glucosides, benzoxazinoide glucosides, flavone
glycosides and phenylethanoid glycosides in this plant. Kanchapoom
et al. (2001) and megastigmane glycosides Wu
et al. (2003). According to various medical literatures, several
adverse the drug-resistant bacteria and fungal pathogens have further complicated
the treatment of infectious diseases in immune compromised, AIDS and cancer
patients (Diamond, 1991). It is likely that plant extract
showing target sites other than those used by antibiotics will be active against
drug-resistant microbial pathogen. However, very little information is available
on such activity of medicinal plants (Lee et al.,
MATERIALS AND METHODS
Collection of plant material: The plant material (whole plant) of Suaeda
monoica was collected from the Ariankuppam region near Pudhucherry and the
collected plant material was botanically identified and confirmed by Herberia
of C.A.S. in Marine Biology, Annamalai University, Tamil Nadu, India.
Preparation of the extracts: The collected materials (whole plant) were
chopped into small pieces separately, shade-dried and coarsely powdered using
a pulverizor. The coarse powders were subjected to successive extraction with
organic solvents such as petroleum ether, ethyl acetate and methanol by Soxhlet
method. The extracts were collected and distilled off on a water bath at atmospheric
pressure and the last trace of the solvents was removed in vacuo. The
resulted extracts were used for preliminary phytochemical screening, antimicrobial
activities and antioxidant activities.
Preliminary phytochemical screening: All the extracts were subjected
to preliminary phytochemical tests followed by the methods of (Harborne,
1998; Sadasivam and Manickam 1996; Trease
and Evans 1983).
Antimicrobial activities: Petroleum ether, ethyl acetate and methanol
extracts of the selected plant were used to prepare various concentrations such
as 25, 50 and 100 mg mL-1, respectively. These were used for antimicrobial
Test microorganisms: The following bacterial strains and fungal were
used for the screening of antimicrobial activity. All the microbial strains
of human pathogens used were procured from IMTECH, Chandigarh and the procured
microbes are the gram-negative bacteria, viz., Escherichia coli (MTCC
724), Proteus vulgaris (MTCC 426), Pseudomonas aeruginosa (MTCC
741) and Salmonella typhi (MTCC 733), Vibrio parahaemolyticus
(MTCC 451), V. vulnificus (MTCC 1145) and the gram-positive bacteria,
Bacillus subtilis (MTCC 441) and Staphylococcus aureus (MTCC 96),
Streptococcus pnemoniae (MTCC 655) and fungi viz., Aspergillus
flavus, A. fumigatus, A. niger (MTCC 1344) and Candida
albicans (MTCC 227), respectively.
Media used: Nutrient Agar (NA) and Potato Dextrose Agar (PDA) were used
respectively for testing the antibacterial and antifungal activity.
DPPH free radical scavenging activity: DPPH free radical scavenging
activity was carried out by following the methods of Yaushisakono
(1978). The 4.3 mg of DPPH (1, 1-Diphenyl-2-picrylhydrazyl) was dissolved
in 3.3 mL methanol; it was protected from light by covering the test tubes with
aluminum foil. The 150 μL DPPH solution was added to 3 mL methanol and
absorbance was taken immediately at 516 nm for control reading. The test sample
of 20 μL was taken and the volume was made uniformly to 150 μL using
methanol. Sample was then further diluted with methanol up to 3 mL and to that
150 μL DPPH was added. Absorbance was taken after 15 min at 516 nm using
methanol as blank on UV-visible spectrometer Systronics, India. Percentage of
inhibition was calculated by using the equation given below:
Fe2+ chelating assay: The chelating activity of the extracts
for ferrous ions Fe2+ was measured according to the method of Dinis
et al. (1994). To 0.5 mL of extract, 1.6 mL of deionized water and
0.05 mL of Fecl2 (2 mM) was added. After 30 sec, 0.1 mL ferrozine
(5 mM) was added. Ferrozine reacted with the divalent iron to form stable magenta
complex species that were very soluble in water. After 10 min at room temperature,
the absorbance of the Fe2+-ferrozine complex was measured at 562
nm. The chelating activity of the extract for Fe2+ was calculated
Preliminary phytochemical screening (qualitative analysis): The results
of preliminary phytochemical screening of Suaeda monoica extracts are
given in the Table 1. Phenolic groups are present in all the
extracts viz., petroleum ether, ethyl acetate and methanol. Steroids and triterpenes
are present both in ethyl acetate and methanol extracts but absent in petroleum
ether extract. Tannins are present only in methanol extract but absent in petroleum
ether and ethylacetate extracts. Carbohydrates and aminoacids are present only
in petroleum ether extract but not in other extracts. Moreover, alkaloids, anthraquinones,
catechins, flavonoids, gum, oils and resins and saponins are completely absent
in all extracts.
||Preliminary phytochemical screening of various extracts of
Suaeda monoica Forssk. ex Gmel
|+: Present, -: Absent
||Quantitative estimation of phytochemicals in Suaeda monoica
||Antimicrobial activity of various extracts of Suaeda monoica
against various microorganisms
|Cl: Clotrimaxazole, A: Ampicillin, C: Ciprofloxacin, K: Kanamycin,
Quantitative assay: The results of quantitative analysis of phytochemicals
in Suaeda monoica are given in the Table 2. In quantitative
analysis, total phenolic group estimated was recorded as 49.1 mg GAE g-1
and total tannins 36.6 mg TAE g-1 at 50 mg concentration.
Antimicrobial activity: In the present study, all the three extracts
tested against various human pathogens are given in the Table
3 and Fig. 1-3. In the present study,
the petroleum ether extract showed the maximum zone of inhibition as 25 and
20 mm against the gram negative bacteria P. aeruginosa, V. parahaemolyticus
and 20 mm against gram positive bacteria B. subtilis at 100 mg mL-1
concentration and moderate zone of inhibition showed as 15 mm against gram negative
bacteria V. parahaemolyticus and 15 mm against gram positive bacteria
S. aureus at 50 mg mL-1 concentration.
||Antimicrobial activity of petroleum ether extract of Suaeda
monoica Forssk. ex Gmel against various microorganisms
||Antimicrobial activity of ethylacetate extract of Suaeda
monoica Forssk. ex Gmel against various microorganisms
||Antimicrobial activity of methanol extract of Suaeda monoica
Forssk. ex Gmel against various microorganisms
||Radical scavenging activity of various extracts of Suaeda
At the same time, the minimum zone of inhibition as 13, 12 and 11 mm against
gram negative bacteria P. vulgaris and S. typhi at 100, 50 and
25 mg mL-1 concentrations. This extract did not show any activity
against the tested gram negative bacteria E. coli and V. vulnificus
and gram positive bacteria S. pneumoniae. This extract did not show any
activity against the tested such as A. flavus, A. fumigatus,
A. niger and C. albicans.
Radical scavenging activity: The results of antioxidant activity are
given in the Table 4. In DPPH activity, methanol extract showed
high percentage of inhibition as 71.4 followed by 42.8 for ethyl acetate extract
and 28.5 for petroleum ether extract, respectively. In Fe2+ chelating
activity, methanol extract showed high percentage of inhibition as 52 followed
by ethylacetate as 46.7 and petroleum ether extract as 40. For antioxidant activity,
the methanol extract showed maximum percentage of inhibition at 50 mg mL-1
concentration and it revealed that the reducing properties are associated with
the presence of chemicals in it.
Generally salt tolerance plant have more amount of phytochemical specially
in phenolic compound. In our result point out phenolics compound pressed in
all the extract (methal, petroleum ether ethyl acetate) (Table
1). Previous studies have reported that salt mash plant. Miftakhova
et al. (1999) reported the phytochemical composition such as amino
acid composition and quantitative character of the plant moisture content, carbohydrate
and flavonoids of the plant, Suaeda physophora belongs to the family
An et al. (2008) isolated 4 compounds such
as methyl 3, 5-di-o-caffeoyl quinate, 3, 5-di-o-caffeoyl quinic acid, isorhamnetin
3-o-β-D-galactoside and quercetin 3-o-β-D-galactoside from methanol
extract of Suaeda glauca and also studied in vitro hepato protective
activity for these isolated compounds. Banerjee et al.
(2008) reported an antioxidant activity and total phenolics of 23 extracts
of some selected mangroves associate plants including Suaeda maritima. So,
in our result indicated moderated compre to previous studies.
Antimicrobial properties of substances are desirable tools in the control of
undesirable microorganisms especially in the treatment of infections diseases.
The active components usually interfere with growth and metabolism of microorganisms
in a negative manner (Aboaba et al., 2006).
||Antifungal activity of Suaeda monoica, (a) S. pneumoniae
(methonal) (b) A. niger, (methonal) (c) A. fumigatus (petroleum
ether) and (d) A. fumigatus (methonal)
In the present study, petroleum ether extract showed the maximum zone of inhibition
against P. aeruginosa, V. parahaemolyticus and B. subtilis
(Fig. 1, 4 and 5), ethyl
acetate extract showed the maximum zone of inhibition against P. aeruginosa
S. aureus and S. pneumoniae and against the fungi A. flavus,
C. albicans, respectively (Fig. 2, 4
and 5) and methanol extract showed the maximum zone of inhibition
B. subtilis, S. aureus, S. pneumoniae and against fungi A. fumigatus,
respectively. (Fig. 3-5). This may be due
to the presence of phenolic groups. Steroids and triterpenes are present both
in ethyl acetate and methanol extracts but absent in petroleum ether extract.
In our study, the maximum zone of inhibition against B. subtilis, S.
aureus may be due to the presence of secondary metabolites such as phenolic
groups as suggested by previous reports by Pereira et
al. (2007). This result mention that moderate compare to (Chandrasekaran
et al., 2009). The significant activity of the results against the
fungi, Candida albicans provides additional confirmation to the phenolic
compounds which are more effective in higher concentration inhibited the growth
of all fungi (Winkelhausen et al., 2005; Pereira
et al., 2007). Antimicrobial activity of salt marsh and coastal medicinal
plants against human pathogenic microorganisms was reported by Ramanathan
(2000) and Sithragaboopathy (2003). For marine drug
discovery, antimicrobial and phytochemical investigation of medicinal mangroves
and other coastal flora reported by Latha (2005).
||Antibacterial activity of Suaeda monoica,
(a) V. parahaemolyticus (petrolium ether), (b) C. albicans
(methonal), (c) S. aureus (methonal) and (d) B. subtilis (methonal)
Even in hospitals, majority of disinfectants such as phenols, lysol, cresols
used are belongs to phenolic groups. The present findings of antimicrobial activity
against the fungi A. flavus, A. fumigatus, A. niger and
C. niger revealed its medicinal potential as fungicides to develop new
leads against fungal infections. Thus from our findings, it was concluded that
the active principles responsible for an antimicrobial activity against these
tested microorganisms should be isolated and identified to develop a new lead
of therapeutic interest.
The antioxidant activity of different extracts may depend on the presence of
polyphenols as reported by Jayaprakasha et al. (2003)
and Ozkan et al. (2005), respectively. Thus
the plants Suaeda monoica possess the property of antimicrobial and antioxidant
property to develop a new of therapeutic interest. This result suggested that
many previous study (Rhee et al., 2009; Thirunavukkarasu
et al., 2011). In our previous study DPPH radical scavenging activity
of Suaeda monoica extract was higher than Sesuvium portulcastrum
and among the other coastal medicinal plants (Thirunavukkarasu
et al., 2010), so in our result remain indicate that suggestion.
We recommended that it would be a good antioxidant drug. From our findings,
it was concluded that the bioactive principles responsible for the antimicrobial
activity and antioxidant activity should be identified, isolated and elucidate
its structure for new drug development in pharmaceutical industry.
The present investigation is only a beginning in the direction of search for
eco-system natural antimicrobial in our coastal environment future work needs
to include standardization of active fractions, identification of the active
compounds and extensive field testing especially challenge through different
routes for finding out the effective administrative route and commercialization
of the antimicrobials. Further work is needed to identify the compounds which
are responsible for antimicrobials and radical scavenging activities. The information
summarized about the Sueada monoica intended to serve as reference
tool to researchers in all field of ethno pharmacology and natural product
The authors are grateful to the Dean, Director and Dr. T. Ramanathan (Assistant
professor) and his Research scholars, Centre of Advanced Study in Marine Biology,
Faculty of Marine Sciences Annamalai University, for providing all support during
the study period. The second author thanks to the UGC-BSR Fellowship, for financial
support during the period of study.
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