Journal of King Saud University – Science 32 (2020) 842–847

Contents lists available at ScienceDirect

Journal of King Saud University – Science

journal homepage: www.sciencedirect.com

Studies on phytochemical, antioxidant, antimicrobial analysis and

separation of bioactive leads of leaf extract from the selected mangroves
G. Eswaraiah a, K. Abraham Peele a, S. Krupanidhi a, R. Bharath Kumar b, T.C. Venkateswarulu a,⇑
a
Department of Bio-Technology, Vignan’s Foundation for Science, Technology & Research, Vadlamudi, 522213 Guntur, Andhra Pradesh, India
b
School of Life Sciences, Rayalaseema University, Kurnool 518002, Andhra Pradesh, India

a r t i c l e i n f o a b s t r a c t

Article history: In the present study, phytochemical and antimicrobial analysis of selected mangrove species: Suaeda
Received 22 November 2018 nudiflora, Lumnitzera racemosa, Ipomoea tuba and Avicennia alba was performed. The phenol found in
Accepted 17 March 2019 all selected mangroves and tannins found in the species: Suaeda nudiflora, Ipomoea tuba and Avicennia
Available online 18 March 2019
alba and terpenoids present in Lumnitzera racemosa, Ipomoea tuba and Avicenia alba. The steroids and
emodins were not found in all selected mangrove species. The antioxidant analysis proved that maximum
Keywords: inhibitory potential of leaf extract was obtained in Lumnitzera racemosa and minimum scavenging activ-
Mangrove species
ity was found in Suaeda nudiflora. The leaf extract with different solvents was screened for antibacterial
Phytochemical
Medicinal property
activity against pathogenic strains: Micrococcus luteus, Pseudomonas aeruginosa, Staphylococcus aureus,
TLC separation Klebsella pneumonia and Bacillus subtilis. The methanol extracts from Squeeda nudiflora and Ipomoea tuba
Rf values was shown highest zone of inhibition compared to other solvent extracts. The order of antimicrobial
activity expressed as inhibitory zones of methanol > acetone > hexane observed for pathogenic strains.
Further, thin layer chromatography (TLC) studies revealed presence of bioactive compounds in the
extracts of the mangrove species.
Ó 2019 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University. This is an
open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction food supplements and also widely used as folk medicines in some
countries (Rasyid et al., 2016). Mangroves are biochemically unique
Mangroves plants are spread in the regions of inter tidal zones of and produce a wide range of natural products widely used for sev-
tropical and sub tropical area mostly on the eastern coast of conti- eral human remedies (Nurdiani et al., 2012). The mangrove, Avicen-
nents. Mangrove species contain a wide variety of chemical com- nia officinalis contain phytochemicals, anti-diabetic and free radical
pounds that could be used as drugs for many epidemic diseases scavenging activities (Zhou et al., 2018). The leaf extracts of man-
(Bobbarala et al., 2009; Behbahani et al., 2018). Mangrove forests grove species namely Rhizophora mucronatz scvatxe and Sonneratia
near estuarine areas act as a barrier against cyclones and tsunami caseolaris have antimicrobial and antioxidant property (Zhao et al.,
(Abeysinghe, 2010). Mangrove wetland serves as spawning and 2011; Gawali and Jadhav, 2011). The methanol leaf extracts from
nursery grounds for many economically important estuarine fishes Suaeda maritima have phytochemicals namely Saponins, terpenoids,
and migratory birds (Thomas et al., 2017; Haq and Wodeyar, 2002). tannins, alkaloids, steroids and are reported for antimicrobial and
Mangrove plants are used as the potential sources of biologically scavenging activity (Santhi and Sengottuvel, 2016). However, till
active chemicals which have commercial applications in the field now no study reported on antimicrobial and phytochemical studies
of ethno- pharmaceutical sector (Nebula et al., 2013). The man- of some mangrove species such as Suaeda nudiflora, Lumnitzera race-
groves species contain neutraceuticals which are used as traditional mosa, Ipomoea tuba and Avicenia alba. Hence, the present study is
aimed to study the antimicrobial, antioxidant and phytochemical
⇑ Corresponding author at: Vignan’s Foundation for Science, Technology & analysis of selected mangrove species.
Research, Vadlamudi, 522213 Guntur, Andhra Pradesh, India.
E-mail address: [email protected] (T.C. Venkateswarulu).
2. Materials and methods
Peer review under responsibility of King Saud University.

2.1. Sample preparation and photochemical analysis

The mangrove species namely Suaeda nudiflora, Lumnitzera race-

Production and hosting by Elsevier
mosa, Ipomoea tuba and Avicennia alba were collected from

https://doi.org/10.1016/j.jksus.2019.03.002
1018-3647/Ó 2019 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
G. Eswaraiah et al. / Journal of King Saud University – Science 32 (2020) 842–847 843

Nizampatnam sanctuary, Guntur, Andhra Pradesh, India. The man- 3. Results and discussion
grove plant species are washed with tap water and cut into small
fragments and make dried under shade away from sun light. The 3.1. Phytochemical analysis
dried leaves are making into powder and stored in polythene bags.
Ten grams of powdered plant material was mixed with 100 ml of The plants extract contain high medicinal values and these
methanol and stirred for 72 h and then filtered. The solvent was compounds can be used to design and develop pharmaceutical
removed by rotary vacuum evaporation. The extracts were used products. The leaf extract of mangrove species contain different
for phytochemical, Antimicrobial and antioxidants analysis (Sheel phytochemicals and bioactive compounds namely alkaloids, ter-
et al., 2014). penoids, steroids and terpenoids (Saranraj and Sujitha, 2015). The
selected mangrove species were found to be positive for presence
2.2. Antioxidant assay of phenols. The plant species Ipomoea tuba and Avicennia alba con-
tain both tannins and alkaloids. Terpenoids was found in leaf
The antioxidant potential of methanol leaf extracts of selected extracts of Lumnitzera racemosa, Ipomoea tuba and Avicennia alba
mangrove species was determined by standard 1, 1-diphenyl-2- and are absent in leaf extracts of Squeeda nudiflora. Steroids and
picryl hydroxyl (DPPH) free radical activity method. The scaveng- emodins are absent in selected mangrove species. The most com-
ing activity was calculated by using following formula and is mon compounds are Tannins, phenols and terpenoids found in Ipo-
expressed in percent (Reddy and Grace, 2016). moea tuba and Avicennia alba (Table 1). In previous studies
reported that the species: Avicenia marina and Avicennia officinalis
% activity ¼ ½control absorbanceðAÞ Extract absorbanceðBÞ= are rich in phytochemicals like alkaloids, flavonoids, terpenoids
control absorbanceðAÞ 100 and phenol. The Avicenia marina fruit extracts contain saponin
and amino acid but these compounds were absent in Avicenia offic-
where A = control and B = sample. inalis (Ramanathan et al., 2012).

2.3. Antimicrobial analysis 3.2. DPPH radical scavenging activity

The extracts from the mangrove species were prepared using DPPH (2, 2-diphenyl-hydrazyl -hydrate) is a stable free radical
different solvents namely: methanol, acetone and hexane. The which accepts an electron to become a stable molecule. The
quantity, 1 mg of leaf sample was mixed in 1 ml of solvent and methanolic leaf extracts of selected mangrove species showed
then sample extracts were tested against pathogenic species such good free radical scavenging activity. The scavenging activity
as Micrococcus luteus, Staphylococcus aureus, Pseudomonas aeurigi- increases with an increase in concentration of the extract. The
nosa, Bacillus subtilis. Sterile cotton swab dipped into culture of DPPH turns yellow from purple indicate presence of antioxidants
pathogenic microbes and inoculated on the surface agar medium in the selected leaf extracts. The percent of scavenging activity of
and then agar surface allowed for drying for 5 min. Then, the sterile methanol extract of selected mangrove species was calculated
filter paper dipped in solvent extract of mangrove species and and graph was plotted against concentration Vs percent of scav-
antibiotic solution and sterile medium were kept on agar surface. enging activity. The maximum scavenger activity, 95.62% was
All the plates were incubated for 24 to 48 h at 37 °C. The zone of found in the leaf exact of Lumnitzera racemosa at concentration of
inhibition was measured in millimeters (mm) and the experiment 200 mg/ml and the minimum scavenging activity, 37.78% was
was performed with different concentrations of the leaf extract i.e., found in the leaf extract of Suaeda nudiflora at concentration of
25 mg, 50 mg, 100 mg and 200 mg and the antibiotic standard and 50 mg/ml. Ascorbic acid used as standard for the assay and the scav-
control plates are maintained (Mukherjee et al., 2014). enging activity of each sample at different concentration was
showed in the Fig. 1. The residual concentration of DPPH depends
2.4. Compound separation thin layer chromatography (TLC) exclusively on the structure of the phenolic compounds. The acces-
sibility of the radical centre of DPPH to each poly phenol could
TLC was performed for alkaloids, tannins, phenols, emodins, ter- influence the order of the antioxidant power (Loganayaki et al.,
penoids etc. The quantity, 50 gm of powdered plant samples of 2013). The mangroves has rich antioxidant system and enzymatic
mangrove species separately extracted with methanol by subject- defense system of mangrove plants include different endogenous
ing it, to maceration at room temperature for overnight and then enzymes such as superoxide dismutase, catalase, glutathione per-
filtered with Whatmans filter paper. The filtrate consists of phyto- oxidase, glutathione reductase and non-enzymatic antioxidants
chemicals are separated by TLC. The TLC plates are prepared by i.e., ascorbate, tocopherols and phenolic compounds serve as pro-
mixing 25 gm of Silica Gel -G with 50 ml distilled water (1:2) for tectors against a wide variety of environmental stresses in plants
two minutes and then, the suspension was distributed over plate (Thatoi et al., 2014).
and plates were dried in hot air oven at 110 °C for 30 min and then
stored in dry atmosphere further, which are used for spotting of 3.3. Antimicrobial analysis for leaf extracts of Suaeda nudiflora
sample. The samples were prepared by diluting crude extracts of
methanol in all the four selected mangrove species separately Antimicrobial activity of leaf extracts of Suaeda nudiflora was
and then, 1–10 ml samples was applied on TLC plate at 2 cm above performed against the pathogenic bacterial strains namely Micro-
its bottom with capillary tubes (Sonam et al., 2017; Biradar and coccus luteus, Pseudomonas aeruginosa, Staphylococcus aureus, Bacil-
Rachetti, 2013). lus subtilis and Klebsiella pneumonia with different concentrations.
The methanol extracts of leaves of Sqseeda nudiflora showed high-
2.5. Statistical analysis est inhibitory zone at the highest concentration (200 mg) against
Staphylococcus aureus and against Micrococcus luteus and even at
All experiments performed in triplicates and the results were low concentration of 25 mg showed significant inhibitory zones
expressed as mean ± standard deviation (SD) for antimicrobial against Staphylococcus aureus. The acetone extracts of leaf sample
and DPPH assays. Data were analyzed by ANOVA (p < 0.05) with showed the antimicrobial effect against Staphylococcus aureus
data analysis system in MS office Excel 2007. and Micrococcus luteus than other bacterial strains and leaf sample
844 G. Eswaraiah et al. / Journal of King Saud University – Science 32 (2020) 842–847

Table 1
Photochemical screening analysis of selected mangroves.

S.No Name of the bioactive compound Suaeda nudiflora Lumnitzera racemosa Ipomoea tuba Avicennia alba
Tannins + _ _ +
Phenols + + + +
Steroids _ _ _ _
Emodins _ _ _ _
Terpenoids _ + + +
Alkaloids _ _ + +

(+) Indicates presence of phytochemical & () Indicates absence of phytochemical

Fig. 1. The antioxidant activity of mangrove species at different concentrations: (a) 50 mg/mL; (b). 100 mg/mL; (c). 150 mg/mL and (d). 200 mg/mL.

with hexane has not showed significant inhibitory zones (Table 2). chloroform showed higher degree of inhibitory zones against
In previous studies reported that leaf extract of Avicennia officinalis selected human pathogenic bacterial strains (Nayak et al., 2018).
showed higher degree of inhibitory zones against other bacterial
strains (Thatoi et al., 2016). The methanol and acetone extracts 3.4. Antimicrobial analysis for leaf extracts of Lumnitzera racemosa
of Avicennia officinalis showed the positive results against bacterial
strains (Sharief et al., 2014). The crude extract of Suaeda nudiflora The leaf extracts of Lumnitzera racemosa showed high inhibi-
with different solvents like hexane, ethyle acetate, methanol and tory zone with highest concentration (200 mg) against Staphylococ-

Table 2
Antimicrobial analysis of leaf extracts of Suaeda nudiflora against the pathogens.

Pathogenic Zone of inhibition(mm)

strains
Methanol (mg) Acetone (mg) Hexane (mg)
25 50 100 200 25 50 100 200 25 50 100 200
Control – – – – – – – – – – – –
M. luteus 4.76 ± 0.25 6.53 ± 0.47 7.46 ± 0.41 8.73 ± 0.25 2.33 ± 0.15 2.83 ± 0.15 3.73 ± 0.25 4.36 ± 0.15 0.4 ± 0.1 0.83 ± 0.15 1.76 ± 0.25 2.13 ± 0.15
S. aureus 7.03 ± 0.15 8.8 ± 0.2 10.03 ± 0.25 12.86 ± 0.15 0.03 ± 0.15 3 ± 0.1 4 ± 0.2 5.76 ± 0.25 1.5 ± 0.2 1.76 ± 0.25 2.76 ± 0.25 3.8 ± 0.2
P. aeroginosa 3.03 ± 0.15 6.76 ± 0.25 6.73 ± 0.25 9 ± 0.2 0.73 ± 0.25 0.5 ± 0.1 1.9 ± 0.1 3 ± 0.2 0.83 ± 0.15 1 ± 0.2 1.76 ± 0.25 2.33 ± 0.15
B. subtilis 2 ± 0.2 2.33 ± 0.15 2.53 ± 0.15 3.46 ± 0.15 1.96 ± 0.15 2.5 ± 0.2 3.5 ± 0.1 4.46 ± 0.15 2 ± 0.2 1.7 ± 0.3 2.5 ± 0.1 2.96 ± 0.15
K. pneumonia 1.96 ± 0.15 2 ± 0.1 3 ± 0.2 4.76 ± 0.25 1.33 ± 0.15 1.86 ± 0.15 1.76 ± 0.25 2.73 ± 0.25 0.53 ± 0.15 1.93 ± 0.20 2 ± 0.2 2.5 ± 0.2
A. niger 1.8 ± 0.2 3.36 ± 0.15 4.46 ± 0.15 4.96 ± 0.25 1.36 ± 0.15 1.76 ± 0.25 1.96 ± 0.25 2.5 ± 0.2 1.1 ± 0.36 1.86 ± 0.15 2.5 ± 0.2 3.06 ± 0.20
Rhizopus 1.7 ± 0.2 2.46 ± 0.15 2.73 ± 0.20 3.13 ± 0.15 2 ± 0.2 2.66 ± 0.75 3.3 ± 0.2 4 ± 0.2 1.1 ± 0.36 2.36 ± 0.15 3.03 ± 0.25 4.75 ± 0.25
G. Eswaraiah et al. / Journal of King Saud University – Science 32 (2020) 842–847 845

cus aureus and significant inhibitory effect against Bacillus subtilis. of mangrove plant namely Ceriopsde candra was reported for pres-
Acetone extracts of leaf sample represents the inhibitory zones ence of many bioactive compounds which have antibacterial prop-
against Pseudomonas aeruginosa than other bacterial strains. The erties (Ravikumar et al., 2010).
leaf sample with hexane showed minimal antimicrobial effective
incomparision with other solvent extracts (Table 3). Earlier, the
3.6. Antimicrobial analysis for leaf extracts of Avicennia alba
studies from Saad et al., (2011) reported that the methanolic leaf
extracts of Lumnitzera littorea showed the potent inhibitory effect
The methanol extracts from the leaves of Avicennia alba showed
against the pathogenic bacterial strains. But, till now no report
significant inhibitory zones against Micrococcus luteus, Pseu-
found on antibacterial activity of Lumnitzera racemosa leaf extracts
domonas aeruginosa, Bacillus subtilis with concentrations 25 mg,
with methanol, acetone and hexane.
50 mg, 100 mg and 200 mg and showed highest inhibitory zones on
Bacillus subtilis. The acetone extracts of leaf sample showed inhibi-
tory zones against Staphylococcus aureus and least on P.aeroginosa.
3.5. Antimicrobial analysis for leaf extracts of Ipomoea tuba
The hexane extracts of leaf showed significant inhibitory zones
against Staphylococcus aureus and Bacillus subtilis. The solvent leaf
The methanol leaf extract of Ipomoea tuba produced highest
extracts of Avicennia albahas not shown the antimicrobial effect
zone of inhibition against Micrococcus luteus and Pseudomonas
on the A. niger (Table 5). The studies from Haq et al., (2014)
aeruginosa and least inhibitory effect was found against Klebsiella
reported that the ethanolic leaf extract of mangrove, Sonneratia
pneumonia. The findings of study also proved superior results
alba showed the antibacterial activity against pathogenic strains:
against Staphylococcus aureus with different concentrations of
Staphylococcus aureus, Bacillus cereus, Escherichia coli I and Pseu-
25 mg, 50 mg, 100 mg, 200 mg. The acetone leaf extracts showed the
domonas aeruginosa. The methanol leaf extract of marine mangrove
highest inhibitory effect against Micrococcus luteus with concentra-
plant Avicennia marina showed the highest antimicrobial activity
tion of 200 mg and least inhibitory effect was found against M.
against pseudomonas aeruginosa (Thamizharasan and
luteus at the concentration (25 mg). Hexane leaf extracts showed
Anbusaravanan, 2016). Compared to other Avicennia species the
minimal effect against effective inhibitory zones against Staphylo-
leaf extract of Avicennia alba is potential because the significant
coccus aureus and no effect against S. aureus and K. pneumonia
inhibitory effect was found on different pathogenic strains.
and the no effect was found on M. luteus, P.aeroginosa and B. sub-
tilis. Methanol extracts showed higher degree of inhibitory zones
against pathogenic strains than the leaf extracts of Acetone and 3.7. Compound separation by TLC
hexane (Table 4). The previous studies reported that leaf extracts
of Rhizophora mangle showed higher degree of inhibitory zones The colour spots represent presence of bioactive compounds
against selected human pathogenic bacterial strains namely Sta- and pale yellow spots on TCL plates was indicative for antioxidants
phylococcus aureus ,Pseudomonas aeruginosa and Bacillus subtilis from mangrove species. The Rf and hRf values calculated for all sep-
(Cruz et al., 2015). The mangrove species have wide ranges of phy- arated bands (Table 6). The bands pattern on TLC sheet is indicated
tochemicals and bioactive compounds which were used for phar- the presence of similar type of phytochemicals. The major bands
macological activities like antimicrobial treatment. Leaf extracts developed from selected mangrove species have similar band pat-

Table 3
Antimicrobial activity of leaf extracts of Lumnitzera racemosa against the pathogens.

Pathogenic Solvent extract

organisms
Methanol (mg) Acetone (mg) Hexane (mg)
25 50 100 200 25 50 100 200 25 50 100 200
Control – – – – – – – – – – – –
M. luteus 2.16 ± 0.15 3.33 ± 0.15 4.13 ± 0.15 5.8 ± 0.15 1.36 ± 0.15 1.83 ± 0.15 1.83 ± 0.15 2.83 ± 0.15 1.16 ± 0.15 0.9 ± 0.1 2.33 ± 0.15 2.36 ± 0.15
S. aureus 4.13 ± 0.15 6.33 ± 0.15 7.8 ± 0.2 8.83 ± 0.15 3.36 ± 0.15 5.8 ± 0.2 5.8 ± 0.2 7.3 ± 0.2 1.83 ± 0.15 2.3 ± 0.2 3.33 ± 0.15 3.4 ± 0.1
P. aeroginosa 3.16 ± 0.15 3.8 ± 0.2 4.36 ± 0.15 5.8 ± 0.2 2.2 ± 0.2 3.86 ± 0.15 3.86 ± 0.15 5.33 ± 0.15 1.13 ± 0.15 1.86 ± 0.15 2.13 ± 0.15 2.8 ± 0.2
B. subtilis 2.8 ± 0.2 3.8 ± 0.2 4.3 ± 0.2 5.8 ± 0.2 2.16 ± 0.15 4.1 ± 0.1 4.1 ± 0.1 5.33 ± 0.15 1.1 ± 0.1 1.8 ± 0.2 2.13 ± 0.15 2.8 ± 0.2
K. pneumonia 1.16 ± 0.15 2.3 ± 0.2 2.86 ± 0.15 4.8 ± 0.2 2.13 ± 0.15 2.83 ± 0.15 2.83 ± 0.15 3.8 ± 0.2 1.16 ± 0.15 1.3 ± 0.2 2.3 ± 0.2 4.13 ± 0.15
A. niger 0.7 ± 0.2 1.16 ± 0.15 2.3 ± 0.2 3.3 ± 0.2 0.66 ± 0.15 2.16 ± 0.20 2.16 ± 0.20 2.3 ± 0.2 1.36 ± 0.15 1.83 ± 0.15 1.8 ± 0.2 2.53 ± 0.15
Rhizopus 0.63 ± 0.15 1.33 ± 0.15 1.83 ± 0.15 2.8 ± 0.2 0.66 ± 0.15 2.13 ± 0.15 2.13 ± 0.15 2.3 ± 0.2 1.33 ± 0.15 1.8 ± 0.2 2.86 ± 0.15 3.3 ± 0.2

Table 4
Antimicrobial activity of leaf extracts of Ipomoea tuba against the pathogens.

Zone of inhibition (mm)

Pathogenic Solvent extract
organisms
Methanol (mg) Acetone (mg) Hexane (mg)
25 50 100 200 25 50 100 200 25 50 100 200
Control – – – – – – – – – – – –
M. luteus 4.72 ± 0.04 6.03 ± 0.15 8.83 ± 0.15 10.86 ± 0.15 1.10 ± 0.02 3.3 ± 0.2 5 ± 0.1 5.73 ± 0.25 1.0 ± 0.03 2.83 ± 0.15 3.3 ± 0.2 4.36 ± 0.15
S. aureus 4.83 ± 0.15 7.86 ± 0.15 8.83 ± 0.15 10.33 ± 0.15 2.83 ± 0.15 3.36 ± 0.15 4.83 ± 0.15 5.83 ± 0.15 1.86 ± 0.15 2.83 ± 0.15 3.3 ± 0.2 4.96 ± 0.25
P. aeroginosa 3.3 ± 0.2 5.83 ± 0.15 8.83 ± 0.15 11.03 ± 0.15 1.83 ± 0.15 2.4 ± 0.1 3.86 ± 0.15 4.8 ± 0.2 1.33 ± 0.15 1.83 ± 0.15 1.88 ± 0.15 2.8 ± 0.2
B. subtilis 3.36 ± 0.15 3.83 ± 0.15 5.86 ± 0.15 9.83 ± 0.15 1.86 ± 0,15 2.83 ± 0.15 4.8 ± 0.2 5.46 ± 0.15 1.33 ± 0.15 1.86 ± 0.15 2.3 ± 0.2 3.83 ± 0.15
K. pneumonia 2.42 ± 0.20 1.86 ± 0.15 3.8 ± 0.2 4.76 ± 0.20 1.36 ± 0.15 1.86 ± 0.15 2.83 ± 0.15 4.83 ± 0.15 2.4 ± 0.1 2.83 ± 0.15 3.33 ± 0.15 4.36 ± 0.15
A. niger 0.86 ± 0.15 1.86 ± 0.15 3.03 ± 0.15 3.33 ± 0.15 0.83 ± 0.15 0.86 ± 0.15 1.83 ± 0.15 2.8 ± 0.2 1.3 ± 0.2 1.86 ± 0.15 3.36 ± 0.15 4.86 ± 0.15
Rhizopus 1.33 ± 0.15 2.4 ± 0.1 2.83 ± 0.15 3.330.15 1.83 ± 0.15 2.8 ± 0.15 3.96 ± 0.15 4.3 ± 0.2 0.83 ± 0.15 1.33 ± 0.15 1.4 ± 0.2 2.3 ± 0.2
846 G. Eswaraiah et al. / Journal of King Saud University – Science 32 (2020) 842–847

Table 5
Antimicrobial activity of leaf extracts of Avicennia alba against the pathogens.

Zone of inhibition(mm)
Pathogenic Solvent extract
strains
Methanol (mg) Acetone (mg) Hexane (mg)
25 50 100 200 25 50 100 200 25 50 100 200
Control – – – – – – – – – – – –
M. luteus 3.86 ± 0.15 5.83 ± 0.15 5.83 ± 0.20 7.3 ± 0.2 2.86 ± 0.15 3.3 ± 0.2 4.8 ± 0.2 5.8 ± 0.2 1.86 ± 0.15 2.3 ± 0.2 2.86 ± 0.15 3.36 ± 0.15
S. aureus 4.33 ± 0.15 5.76 ± 0.20 7.83 ± 0.15 9.33 ± 0.15 3.3 ± 0.2 3.5 ± 0.43 5.86 ± 0.15 6.83 ± 0.15 1.33 ± 0.15 1.86 ± 0.15 3.83 ± 0.15 4.3 ± 0.2
P. aeroginosa 3.8 ± 0.2 5.86 ± 0.15 6.83 ± 0.15 7.3 ± 0.2 1.86 ± 0.15 3.33 ± 0.15 4.3 ± 0.2 5.83 ± 0.15 1.86 ± 0.15 2.3 ± 0.2 2.36 ± 0.15 3.36 ± 0.15
B. subtilis 4.83 ± 0.15 0.86 ± 0.15 6.8 ± 0.2 8.83 ± 0.15 2.13 ± 0.75 5.03 ± 0.15 5.3 ± 0.2 6.83 ± 0.15 2.36 ± 0.15 2.86 ± 0.15 3.8 ± 0.2 4.76 ± 0.20
K. pneumonia 1.03 ± 0.15 1.3 ± 0.2 1.83 ± 1.03 5.33 ± 0.15 1.86 ± 0.15 2.33 ± 0.15 3.3 ± 0.2 3.83 ± 0.15 1.33 ± 0.15 1.36 ± 0.15 2.76 ± 0.15 3.33 ± 0.15
A. niger 1.83 ± 0.15 2.4 ± 0.1 3.36 ± 0.15 3.8 ± 0.2 1 ± 0.1 1.83 ± 0.15 2.33 ± 0.15 2.83 ± 0.15 0.5 ± 0.1 1.33 ± 0.15 1.86 ± 0.15 2.83 ± 0.15
Rhizopus 2.36 ± 0.15 2.83 ± 0.15 3.76 ± 0.15 5.86 ± 0.15 1.83 ± 0.15 2.36 ± 0.15 2.8 ± 0.2 4.86 ± 0.15 0.83 ± 0.15 2.33 ± 0.15 2.83 ± 0.05 3.86 ± 0.15

Table 6
The Rf and hRf values of the separated bands for the leaf extracts of selected mangrove species on TLC sheet.

Number of similar bands S. nudiflora L. racemosa I. tuba A. alba

Rf hRf Rf hRf Rf hRf Rf hRf
01 0.38 ± 0.005 39.33 ± 0.57 0.35 ± 0.005 36 ± 1 0.36 ± 0.01 35.33 ± 0.57 0.43 ± 0.005 43.66 ± 0.57
02 0.43 ± 0.007 43.33 ± 0.37 0.38 ± 0.005 38.33 ± 1 0.42 ± 0.01 42 ± 0.03 0.64 ± 0.01 64 ± 1
03 0.64 ± 0.005 64.33 ± 0.41 0.7 ± 0.01 70 ± 0.03 0.48 ± 0.01 48 ± 1 0.72 ± 0.01 72 ± 0.24
04 0.7 ± 0.01 70 ± 1 0.83 ± 0.005 83.66 ± 1 0.7 ± 0.01 70 ± 1 0.86 ± 0.01 86 ± 0.02
05 0.86 ± 0.005 86.33 ± 0.57 0.86 ± 0.01 86 ± 0.04 0.85 ± 0.57 85.33 ± 0.57 0.89 ± 0.01 89 ± 1

Fig. 2. The compound in methanol leaf extract of mangrove species separated on TLC sheet: (a). Detection of bands at 254 nm, (b). Detection of bands at 366 nm & (c).
Detection of bands after spray with Anisaldehyde sulphuric acid reagent (ANS).

tern (Fig. 2). The yellow color spots on TLC sheet represent the iflora. The TLC study revealed the presence of the bioactive mole-
presence of bioactive compounds and antioxidants. Hanna et al. cules with scavenging activity. Hence, the study proved that the
in (2008) reported that the yellow colour spots indicate presence mangrove extracts used for development of novel bioactive mole-
of carotenoids and other phenolic compounds. The phenolic com- cules for therapeutic applications and further studies in loop for
pounds from plants are known to be good anti-oxidants. selective isolation and identification of bioactive molecules of
mangrove plants.
4. Conclusion
Conflict of interest
The leaf extracts of mangroves species selected for the study
reveals the presence of phytochemicals and among the four sam- The authors declare that they have no conflict of interest.
ples, potent antioxidant activity is found in the leaf extract of Lum-
nitzera racemosa. The methanolic leaf extract from the selected Acknowledgements
mangroves showed highest antimicrobial activity in comparison
with other solvent extracts and the highest the zone of inhibition Authors acknowledge the facilities supported by DST-FIST and
found on Staphylococcus aureus from the leaf extract of Suaeda nud- VFSTR.
G. Eswaraiah et al. / Journal of King Saud University – Science 32 (2020) 842–847 847

References Rasyid, A., A.S. M.A., Nurdin, N., Jaya, I., 2016. Impact of human interventions on
mangrove ecosystem in spatial perspective. In IOP Conference Series: earth and
environmental science. IOP publishing 47, 012–041.
Abeysinghe, P.D., 2010. Antibacterial activity of some medicinal mangroves against
Ravikumar, S., Gnanadesigan, M., Suganthi, P., Ramalakshmi, A., 2010. Antibacterial
antibiotic resistant pathogenic bacteria. Indian J. Pharm. Sci. 72, 167–172.
potential of chosen mangrove plants against isolated urinary tract infections
Behbahani, B.A., Yazdi, F.T., Shahidi, F., Noorbakhsh, H., Vasiee, A., Alghooneh, A.,
bacterial pathogens. Int. J. Med. Med. Sci. 2, 94–99.
2018. Phytochemical analysis and antibacterial activities extracts of mangrove
Reddy, A.R.K., Grace, J.R., 2016. In vitro evaluation of antioxidant activity of
leaf against the growth of some pathogenic bacteria. Microb. Pathog. 114, 225–
methanolic extracts of selected mangrove plants. Med. Aromat. plants. 5, 2167–
232.
10412.
Biradar, S.R., Rachetti, B.D., 2013. Extraction of some secondary metabolites & thin
Saad, S., Taher, M., Susanti, D., Qaralleh, H., Rahim, N.A.B.A., 2011. Antimicrobial
layer chromatography from different parts of Centellaasiatica L (URB). Am. J. Life
activity of mangrove plant (Lumnitzera littorea). Asian Pac. J. Trop. Biomed. 4,
Sci. 1, 243–247.
523–525.
Bobbarala, V., Vadlapudi, V.R., Naidu, C., 2009. Antimicrobial potentialities of
Santhi, K., Sengottuvel, R., 2016. Qualitative and quantitative phytochemical
mangrove plant Avicennia marina. J. Pharm. Res. 2, 1019–1021.
analysis of Moringa concanensis Nimmo. Int. J. Curr. Microbiol. App. Sci. 5,
Cruz, S.M., Marroquin, N., Alvarez, L.E., Chang, D.E., Caceres, A., 2015. Evaluation of
633–640.
Mangrove (Rhizophora mangle L.) products as coloring, antimicrobial and
Saranraj, P., Sujitha, D., 2015. Mangrove medicinal plants: a review. Am.–Eurasian J.
antioxidant agents. Int. J. Phytocosmetics Nat. Ingredients 2, 1–7.
Toxicol. Sci. 7, 146–156.
Gawali, P., Jadhav, B.L., 2011. Antioxidant activity and antioxidant phytochemical
Sharief, M.N., Srinivasulu, A., Satyaveni, P., Umamaheswararao, V., 2014.
analysis of mangrove species Sonneratia alba and Bruguiera cylindrica. Asian J.
Quantification of phytochemicals and antibacterial activity of fruit extract of
Microbiol. Biotechnol. Environ. Exp. Sci. 13, 257–261.
Avicennia officinalis. Asian. J. Pharm. Clin. Res. 7, 127–130.
Hanna, H., El-Baky, Abd., Farouk, K.E., Gamal, S.E., Baroty, 2008. Evaluation of
Sheel, R., Nisha, K., Kumar, J., 2014. Preliminary phytochemical screening of
marine algae Ulvalactuna Las a source of natural preservative ingredient. Am.-
methanolic extract of Clerodendronin fortunatum. IOSR.J.Appl. Chem. 7,
Eur. J. Agric. Environ. Sci. 3, 434–444.
10–13.
Haq, I., Hossain, A.S., Moneruzzaman, M., Amir, K., Merican, F., Faruq, G., Azirun, M.
Sonam, M., Singh, R.P., Pooja, S., 2017. Phytochemical screening and TLC profiling of
S., 2014. Antioxidant and antibacterial activities of different extracts and
various extracts of Reinwardtia indica. Int. J. Pharmacognosy Phytochem. Res.,
fractions of a mangrove plant Sonneratia alba. Int. J. Agric. Biol. 16, 706–714.
523–527
Haq, K.M.F., Wodeyar, A.K., 2002. An ecological study of habitat of mangrove forest
Thamizharasan, S., Anbusaravanan, N., 2016. Antibacterial potential of
of Bangladesh. J. Human Ecol. 13, 225–230.
mangrove plant Avicennia marina against a clinical pathogen. Int. J. Zool. Stud.
Loganayaki, N., Siddhuraju, P., Manian, S., 2013. Antioxidant activity and free radical
1, 14–16.
scavenging capacity of phenolic extracts from Helicteres isora L. and Ceiba
Thatoi, H., Samantaray, D., Das, S.K., 2016. The genus Avicennia, a pioneer group of
pentandra L. J. Food Sci. Technol. 50, 687–695.
dominant mangrove plant species with potential medicinal values: a review.
Mukherjee, N., Sutherland, W.J., Dicks, L., Huge, J., Koedam, N., Dahdouh-Guebas, F.,
Front. Life Sci. 9, 267–291.
2014. Ecosystem service valuations of mangrove ecosystems to inform decision
Thatoi, H.N., Patra, J.K., Das, S.K., 2014. Free radical scavenging and antioxidant
making and future valuation exercises. PloS one 9, e107706.
potential of mangrove plants: a review. Acta physiologiae plantarum. 36, 561–
Nayak, B., Roy, S., Roy, M., Mitra, A., Karak, K., 2018. Phytochemical, antioxidant and
579.
antimicrobial screening of Suaeda maritima L (Dumort) against human
Thomas, N., Lucas, R., Bunting, P., Hardy, A., Rosenqvist, A., Simard, M., 2017.
pathogens and multiple drug resistant bacteria. Indian J. Pharmaceutical Sci.
Distribution and drivers of global mangrove forest change, 1996–2010. PloS one
80, 26–35.
12, e0179302.
Nebula, M., Harisankar, H.S., Chandramohanakumar, N., 2013. Metabolites and
Zhao, K., Penttinen, P., Guan, T., Xiao, J., Chen, Q., Xu, J., Strobel, G.A., 2011. The
bioactivities of Rhizophoraceae mangroves. Nat. Products Bioprospecting 3,
diversity and anti-microbial activity of Endophytic actinomycetes isolated from
207–232.
medicinal plants in Panxi plateau, China. Curr. Microbiol. 62, 182–190.
Nurdiani, R., Firdaus, M., Prihanto, A.A., 2012. Phytochemical screening and
Zhou, J., Diao, X., Wang, T., Chen, G., Lin, Q., Yang, X., Xu, J., 2018. Phylogenetic
antibacterial activity of methanol extract of mangrove plant (Rhizophora
diversity and antioxidant activities of culturable fungal endophytes associated
mucronata) from porong river estuary. J. Basic Sci. Technol. 1, 27–29.
with the mangrove species Rhizophora stylosa and R. mucronata in the South
Ramanathan, T., Shanmugapriya, R., Renugadevi, G., 2012. Phytochemical
china sea. PloS one 13, e0197359.
characterization and antimicrobial efficiency of mangrove plants Avicennia
marina and Avicennia officinalis. Int. J. Pharmaceutical Biol. Archive 3, 348–351.