Journal of Cluster Science (2019) 30:1103–1113

https://doi.org/10.1007/s10876-019-01571-2(0123456789().,-volV)(0123456789().
,- volV)

ORIGINAL PAPER

Green Synthesis of Sliver Nanoparticles Using Avicennia officinalis

and Xylocarpus granatum Extracts and In vitro Evaluation
of Antioxidant, Antidiabetic and Anti-inflammatory Activities
Swagat Kumar Das1 • Supriti Behera1 • Jayanta Kumar Patra2 • Hrudayanath Thatoi3

Received: 8 March 2019 / Published online: 27 April 2019

 Springer Science+Business Media, LLC, part of Springer Nature 2019

Abstract
The present study involves biosynthesis of AgNPs using Avicennia officinalis and Xylocarpus granatum mangrove plants
along with evaluation of their potential biomedical applications. The synthesized AgNPs were characterized by UV–Vis
spectroscopy, FTIR analysis, scanning electron microscope, particle size analyzer, X-ray diffraction (XRD). The syn-
thesized AgNPs showed absorption maxima at 470 nm for X. granatum (XG-AgNPs) and 420 nm for A. offcinalis (AO-
AgNPs) which corresponds to their respective surface Plasmon resonance. The FTIR analysis reveals capping of phenolic
groups providing stability of synthesized AgNPs. The morphology of silver nanoparticles was confirmed by SEM tech-
nique. The dynamic light scattering study (DLS) also confirmed the size distribution of synthesized AgNPs. XRD peaks at
2h range of 20–70o corresponds (111), (200) and (220) reflection planes indicating the structure of metallic silver. AO-
AgNPs exhibited better DPPH scavenging, superoxide and protein denatuaration activity with IC50 values of 0.14, 0.32 and
0.21 mg/ml. However, XG-AgNPs exhibited better a-amylse and a-glucosidase inhibition potential as compared to AO-
AgNPs. It could be concluded that X. granatum bark extracts and A. officinalis leaf extract can be used efficiently for the
synthesis of biologically active silver nanoparticles which could be exploited pharmaceutical applications.

Keywords Silver nanoparticle  Mangrove  Antioxidant  Antidiabetic  Anti-inflammatory

Introduction biomedical properties such as antimicrobial, wound healing

ointments, also for food packaging, medical instruments,
Nanobiotechnology is presently one of the most dynamic textile coatings, cosmetics etc. [2]. The conventional
disciplines of research in contemporary science whereby techniques used for synthesis of AgNPs such as laser
plants and different plant products are finding an impera- ablation, gamma irradiation, electron irradiation, photo-
tive use in the synthesis of nanoparticles (NPs) [1]. chemical methods, and microwave processing are either
Amongst the different NPs, the silver nanoparticle expensive, or labour intensive. Further, chemical methods
(AgNPs) have demonstrated improved electrical conduc- usually involves toxic or hazardous solvents and reductants
tivity, chemical stability, catalytic activity, and distinct making these methods unsuitable [3]. Therefore, develop-
ment of simple and environmentally friendly alternative
approaches for preparation of nanoparticles using non-toxic
& Swagat Kumar Das reagents and cost effectively is highly desirable. In this
das.swagat@gmail.com context, green synthesis of nanoparticles using microbes,
1
marine organisms, and plant extracts have become popular
Department of Biotechnology, College of Engineering and
as these methods are biocompatibile, environmental
Technology, Ghatikia, Kalinga Nagar, Bhubaneswar,
Odisha 751003, India friendly and economic. Amongst the different approaches
2 of greener syntheses of silver nanoparticles, plant mediated
Research Institute of Biotechnology and Medical Converged
Science, Dongguk University, Seoul, Republic of Korea synthesis of AgNPs is considered as a widely accept-
3 able cost-effective, environmentally friendly alternate
Department of Biotechnology, North Odisha University,
Baripada, Odisha 757003, India technology for rapid production of AgNPs. Although, the

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1104 S. K. Das et al.

potential of higher plants as source for this purpose is still Green Synthesis of Silver Nanoparticle (AgNPs)
largely unexplored. Recently, synthesis of nanoparticles
using mangrove plants has recently got attention. The The silver nanoparticles (AgNPs) were synthesized fol-
mangrove plants are woody, specialized types of trees lowing the method of Rao and Savithramma [6] with some
growing in brackish wetlands in the tropical and sub- modification. Briefly, the leaf extracts of A. officinalis and
tropical inter-tidal coastal zones and river deltas. These bark extracts of X. granatum and AgNO3 solution (10 mM)
mangrove plants possess different class of bioactive sec- were taken in separated conical flask in 1:19 ratio and
ondary metabolites such as alkaloids, glycosides, ter- allowed to stand for 6 h at room temperature. The reduc-
penoids, flavonoids, and other poyphenols that can be tion of silver nitrate to AgNPs was confirmed by change in
exploited for various therapeutic needs such as antimicro- colour of the solution. The extract contents were then
bials, antioxidants, anti-inflammatory, antidiabetic, cyto- centrifuged at 10,000 rpm for 20 min. The Ag nanoparti-
toxic etc. [4]. The rich content of polyphenols in the cles were washed with distilled water for 2–3 times to
mangrove plants can be exploited as these polyphenols are remove any unbound phytoconstituents. Then the prepared
reported to be strong reducing agents and also have a AgNPs were dried at 50 C in a dry oven and stored at 4 C
tendency to adsorb on the surface of NPs and also func- till further use.
tionalize the resultant NPs [5].
Considering the vast potentiality of mangrove plants as Characterization of Nanoparticles
source of reducing agents for nanoparticle synthesis, the
present work aims to apply a biological green technique for The mangrove plants mediated synthesized AgNPs were
the synthesis of silver nanoparticles as an alternative to characterized by different analytical techniques such as,
conventional methods. In this regard, leaf extract of Avi- UV–visible spectroscopy, Fourier transformation infrared
cennia officinalis L. and bark extract of Xylocarpus spectroscopy (FT-IR), dynamic light scattering (DLS) and
granatum J. Koenig (www.theplantlist.org) two medici- X-ray Powder diffraction (XRD).
nally important mangrove plants were used for synthesis of
silver nanoparticles (AgNPs). The green synthesized UV–visible Spectral Analysis
AgNPs were characterized by several techniques such as
UV–Vis, FT-IR, DLS and XRD, Moreover,the antioxidant, The UV–Vis spectrophotometer UV–117 (SystronicsTM)
antidiabetic and anti-inflammatory properties of the syn- was used to measure UV–Vis absorbance spectra of AgNPs
thesized AgNPs were evaluated. synthesized by A. officinalis leaf and X. granatum bark
extracts. The absorbance was measured in 400–800 nm
range with a 1 nm step and the characteristic peaks were
Materials and Methods detected. The spectra were recorded at the intervals of
1 min to 10 min and the distilled water was used as a
Collection of Mangrove Plants and Extraction baseline.

The leaves of Avicennia officinalis L. and barks of Xylo- FT-IR Analysis

carpus granatum J. Koenig mangrove plants were collected
from Mahanadi Delta mangrove forest of Odisha. The plant The interactions of A. officinalis leaf and X. granatum bark
specimens were identified by Prasanna Kumar Nayak, extracts and AgNPs were analyzed with FTIR spec-
Herbarium keeper, Integrated Coastal Zone Management troscopy. For FTIR measurements, sample ground with
Project (ICZMP), Forest Department, Govt. of Odisha. KBr and pellet was analyzed using an FTIR spectropho-
After collection, the plant samples were cleaned in tap tometer in the range of 400–4000 cm-1.
water, air dried under shade at room temperature cut into
small pieces and homogenized into a fine powder using SEM Analysis
mechanical grinder. About 10 g of powdered leaf sample
of A. officinalis and bark sample X. granatum were taken The field-emission scanning electron microscopy (FESEM)
separately in two 500 ml conical flask containing 100 ml was used to study the surface morphology of dried samples
of sterile distilled water and was boiled for 20 min. After of synthesized silver nanoparticles (JEOL, JSM7600F).
cooling the extracts were filtered through Whatman No.1
filter paper and the filtrate was evaporated to dryness. The
dried leaf and bark extracts of A. officinalis and bark
sample X. granatum respectively were used for the syn-
thesis of silver nanoparticle.

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Green Synthesis of Sliver Nanoparticles Using Avicennia officinalis and Xylocarpus granatum… 1105

DLS Analysis
where DOD = Abs after 15 min - Abs at 0 min
The dynamic light scattering (DLS) is used to characterize
Antidiabetic Study
the surface charge and the size distribution of the AgNPs
suspended in a liquid [7]. The AgNP solutions after fil-
a-Amylase Inhibition Assay
tration were transferred to the cuvette and the hydrody-
namic radius was measured in the particle analyzer
Amylase activity was assayed using chromogenic 3,5-
(Malvern). All measurements were performed at room
dinitrosalicylic acid (DNS) method [11]. AgNPs of dif-
temperature.
ferent concentrations (0.1 mg/ml to 0.5 mg/ml) were
allowed to incubate with 200 ll of the a-amylase enzyme
XRD Analysis
and 100 ll of 2 mM phosphate buffer (pH 6.9) for 5 min
followed by addition of 0.5% starch solution. After incu-
XRD is used for the phase identification and characteri-
bation for 5 min at 37 C, 400 ll of reaction mixture was
zation of the crystal structure of the silver nanoparticles
removed and then 200 ll of dinitrosalicylic acid reagent
[8]. X-rays penetrate into the nanomaterial and the result-
was added and were boiled for 15 min at 90 C. The
ing diffraction pattern is compared with standards to obtain
reaction mixture was then diluted with 1.8 ml of distilled
structural information. The crystalline phase of A. offici-
water. The absorbance was recorded at 540 nm. The % of
nalis leaf and X. granatum bark extracts mediated syn-
enzyme inhibition was calculated.
thesized Ag nanoparticles were determined by X-ray
diffraction (XRD) using Cu–Ka radiations (k 1.5406 Å)
a-Glucosidase Inhibitory Activity
in 2h range from 20 to 70. The XRD method is suitable to
determine the crystal structures by analyzing the positions
The a-glucosidase was assayed following the method of
and intensities of diffraction peaks.
Apostolidis et al. [12]. a-Glucosidase was premixed with
AgNPs at various concentrations (0.1 mg/ml and 0.5 mg/
Bioactivity Studies ml) and incubated at 37 C for 10 min. 3 mM PNPG in
0.2 M sodium phosphate buffer (pH 7.4) as substrate was
Antioxidant Study
added to the reaction mixture to start the reaction. The
reaction was incubated at 37 C for 20 min and stopped by
DPPH Scavenging Assay The free radical scavenging
adding 2 mL of Na2CO3. The a-glucosidase activity was
activity of the synthesized AgNPs on the stable radical 1,1-
determined by measuring the p-nitrophenol release from
diphenyl-2-picrylhydrazyl was estimated (DPPH) [9].
pNPG at 405 nm. The percentage inhibition of enzyme
Synthesized AgNPs particles at different concentrations
activity was calculated.
(0.1 mg/ml to 0.5 mg/ml) were incubated with 3.0 ml of a
DPPH methanol solution (0.1 mM) in dark for 30 min at
Anti-inflammatory Assay
room temperature followed by measuring the absorbance at
517 nm.
The protein denaturation activity study was carried fol-
lowing the method of Dey et al. [13]. Briefly, the reaction
Superoxide Scavenging Activity The superoxide scav-
mixture consisting of 0.2 ml of egg albumin (from fresh
enging was determined by the nitro blue tetrazolium
hen’s egg), 2.8 ml of phosphate buffered saline (PBS, pH
reduction method [10]. The reaction mixture containing
6.4) and AgNPs of varying concentrations (0.1 mg/ml and
ethylene diaminetetraacetic acid (EDTA) (0.1 mM), nitro
0.5 mg/ml) were incubated at 37 C for 15 min followed
blue tetrazolium (1 mM), Na2CO3 (50 mM) and various
by heating at 70 C for 5 min. After cooling, their absor-
concentrations of synthesized AgNPs (0.1 mg/ml to
bance was measured at 660 nm. The percentage inhibition
0.5 mg/ml) were taken. About 1.5 ml of (3 mM) of
of protein denaturation was calculated by using the fol-
hydroxylamine hydrochloride was added to initiate the
lowing formula:
reaction. The absorbance was measured at the start of the
reaction and after an interval of 15 min at 560 nm. The Percentage of inhibition ¼ 100  ½Vt =Vc  1
control was simultaneous run without plant extract. where Vt = absorbance of test sample, Vc = absorbance of
% of superoxide inhibition control.
DOD of control  DOD of treated sample
¼  100
DOD of control

123
1106 S. K. Das et al.

Statistical Analysis of circumstances, no colour transformation was noticed in

the absence of plant extract. The colour change arises due
All data were presented as means ± standard deviations to excitation of surface plasmon resonance (SPR) in syn-
(SD) where appropriate. The means of all the parameters thesized AgNPs [1] and corroborated with the earlier
were examined for significance by analysis of variance studies [14, 15]. The visual change in colour of the solu-
(ANOVA) and in case of significance mean separation tions was different for both A. officinalis and X. granatum
were accomplished by SPSS (Edition 20) statistical soft- extracts may be due to the presence of different phyto-
ware package. Differences were considered significant at a components present in them which causes a varied range of
level of p \ 0.05 with Bonferroni post hoc test. bioreduction. Variation in the availability of phyto-
molecules in two different mangrove plant extracts is
responsible for this difference in the absorbance values.
Results and Discussion
UV–Vis Spectral Analysis
The plants are an excellent source of important secondary
metabolites of medicinal importance compared to bacteria The formation of AgNPs was further confirmed by UV–Vis
and fungi, their by providing greater scope for the pro- spectral study, which is an authentic technique to monitor
duction of biocompatible nanoscale particles. The present the progress of the reaction during the reduction of Ag ?
study deals with the green synthesis of silver nanoparticle ions. Typically, UV–Vis spectroscopy serves as a prelim-
using extracts of two medicinally important mangrove inary tool to confirm the formation of NPs. The UV–Vis
plants viz. X. granatum and A. officinalis, their character- absorption in the visible light region of 420–450 nm is an
ization and evaluation of their antioxidant, carbohydrate evidence of the presence of surface plasmon resonance
metabolizing enzyme inhibition and anti-inflammatory (SPR) of AgNPs [16]. From the results of UV–Vis
potential. absorption spectra of the synthesized NPs in solution it is
clear that the AgNPs showed their characteristic absorption
Green Synthesis and Characterization of AgNPs maxima between 420 and 490 nm. The present analysis
showed characteristic UV–Vis absorption maxima at
Addition of leaf and bark extracts of A. officinalis and X. 420 nm for AO-AgNPs and 470 nm for XG-AgNPS
granatum to AgNO3 solution resulted in distinct change in (Fig. 2a, b). The difference in the position of absorption
colour of the reaction mixture. The initial yellow AgNO3 peak of AgNPs may be due to the difference in nanopar-
solution turned into dark brownish which indicates the ticle particle size and shape. This observation is in accor-
gradual reduction of AgNO3 by mangrove plant extracts dance with the earlier reports on absorption spectra for
(Fig. 1a, b). The Ag? ion reduction was confirmed by the AgNPs [15, 17].
apparent transformation in the colour. Under a similar set
FT-IR Analysis
a
FT-IR spectroscopic analysis of XG-AgNPs and AO-
AGNPs were carried out to investigate the functional
groups present in X. granatum bark and A. officinalis leaf
extracts which may be responsible for synthsis silver
nanoparticles and surface capping. The FT-IR spectra of
XG-AgNPs shown in Fig. 3a reveals the possible biomo-
Before After lecules present in the bark extract of X. granatum which is
b accountable for the reduction of silver ions. The FT- IR
spectrum of XG-AgNPs shows absorption peaks at
3441.37, 1621.11 and 1384.2 cm-1. The broad band at
3434.59 cm-1 corresponds to the strong stretching vibra-
tions of hydroxyl group (–OH) of phenolic compounds that
suggests the binding of silver ions with hydroxyl group
(OH). The intense bands at 1621.11 can be attributed to
C=C and 1384.20 cm-1 from C–H stretching which may
Before After
be due to ring stretching vibration of heterocyclic com-
Fig. 1 Colour change during synthesis of silver nanoparticles using pound. Similarly, the FT-IR spectra of the AO-AgNPs was
the extracts of a X. granatum and b A. officinalis recorded in order to identify the functional groups present

123
Green Synthesis of Sliver Nanoparticles Using Avicennia officinalis and Xylocarpus granatum… 1107

Fig. 2 UV–Vis spectroscopy

analysis of silver nanoparticles
using the extracts of a X.
granatum and b A. officinalis

in the aqueous extract of A. officinalis leaf involved in the nanoparticles. It has been reported elsewhere that aggre-
synthesis and stabilization of silver nanoparticles (Fig. 3b). gation of nanoparticles can be induced during sample
The FT-IR spectra showed absorption peaks at 3436.88, preparation and evaporation of solvent which may be the
1631.3, 1384.35, 1114.21 and 825.14 cm-1. A broad peak possible reason behind aggregation of XG-AgNPs and AO-
at 3436.88 cm-1 strongly suggests the binding of silver AGNPs [18].
ions with hydroxyl group (OH). The other four bands at
1631.3, 1384.35 and 1114.21 825.14 cm-1 were due to DLS Analysis
C=C, C–H, C–O functional groups. The bioactive com-
pounds present in the X. granatum and A. officinalis DLS is a very efficient and effective tool in analyzing
extracts might have been responsible for the reduction and quantitative size distributions and quantity of monodis-
stability of the silver nanoparticles. persity in colloidal solutions. The differential intensity,
number related to particle size distributions of the A.
SEM Analysis officinalis and X. granatum extracts mediated AgNPs were
obtained from DLS study (Fig. 5a, b). The DLS study
The FESEM study provided further information on surface demonstrated that X. granatum bark mediated silver
morphology of XG-AgNPs and AO-AGNPs. The FESEM nanoparticle (XG-AgNP) particle ranged between 20 and
revealed that both XG-AgNPs and AO-AGNPs are poly- 1000 nm with the average particle intensity at 98.77 nm.
dispersed (Fig. 4a, b). The clusters of silver nanoparticles Similarly, the population of particle for A. officinalis leaf
were also observed which may be due to aggregation of mediated silver nanoparticle (AO-AgNP) varied between

123
1108 S. K. Das et al.

79
a 78

77

76

75

74
%T

73

72

71 1621.11cm-1, 71.53%T

70
1384.20cm-1, 70.45%T

69

68

67 3441.37cm-1, 66.36%T

66
4000 3500 3000 2500 2000 1500 1000 500 400

cm-1

b 78
75

70
825.14cm-1, 72.50%T
1114.21cm-1, 68.98%T 613.68cm-1, 72.56%T
65

60 1631.30cm-1, 63.66%T

55

50
%T

45 3436.88cm-1, 47.30%T

40

35

30

25
1384.35cm-1, 18.78%T
20
17
4000 3500 3000 2500 2000 1500 1000 500 400

cm-1

Fig. 3 FT-IR analysis of silver nanoparticles using the extracts of a X. granatum and b A. officinalis

50 and 1000 nm with the average particle intensity at structure of AgNPs Comparison of the sharp peaks of the
181.4 nm. This observation is in accordance with the ear- obtained data in the present study with the standard,
lier DLS study for AgNPs [14]. clearly indicate the crystalline nature of the synthesized
nanoparticles which is in nano regime and agreement
XRD Analysis with the earlier reports [16, 17]. In the above XRD
profile, apart from normal peaks, some additional and
The X- ray diffraction technique is used to analyze the unknown peaks were also noticed at the vicinity of the
metallic nature of particles. The X-ray diffraction (XRD) characteristic peaks of Ag, which might have resulted
pattern of the X. granatum and A. officinalis extract due to the presence of bioorganic compounds of the
mediated AgNPs are shown in Fig. 6a, b. The XRD extract that act as capping agent in stabilizing the
pattern shows peak in the whole spectrum of 2h values nanoparticle [19].
ranging from 20o to 70o. The pattern of XG-AgNPs
showed three distinct characteristic peaks at 38.09, 46.25 Bioactivity Studies
and 64.55 in 2h range which can be indexed to the (111),
(200) and (220) reflection planes. Similarly, three dis- X. granatum and A. officinalis extract mediated Silver
tinct peaks were observed in AO-AgNPs at 38.21, 44.38 nanoparticles were tested for their possible therapeutical
and 64.58 which were indexed as (111), (200) and (220). potential for antioxidant, anti-diabetic anti-inflammatory
The reflection planes obtained from XG-AgNPs and AO- activities.
AgNPs data obtained were matched with the database of
Joint Committee on Powder Diffraction Standards
(JCPDS file No.04-0783) and were interpreted for the
structure of metallic silver with face-centered cubic

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Green Synthesis of Sliver Nanoparticles Using Avicennia officinalis and Xylocarpus granatum… 1109

demonstrates that AO-AgNPs was effective as compared

with XG-AgNPs in scavenging DPPH radicals as the for-
mer was having an IC50 value of 0.14 mg/ml in comparison
to the later one having 0.25 mg/ml (Table 1). The standard
antioxidant compound BHT under similar condition could
scavenge DPPH radical with IC50 value of 0.04 mg/ml.
Earlier study have shown that another mangrove plant
Sonneratia apetala mediated AgNPs could scavenge DPPH
radical at 65.31% at 100 lg/ml [21]. Similarly, it has also
been reported that AgNPs synthesized using extracts of two
mangrove plants Heritiera fomes and S. apetala could
scavenge DPPH radicals with IC50 values \ 0.2 mg/ml
[14] which is in accordance with the present study.
The antioxidant potential of the synthesized AgNPs was
evaluated in terms of the superoxide radical scavenging
activity also. Superoxide radicals were generated in vitro
chemically by hydroxylamine hydrochloride, which redu-
ces nitro blue tetrazolium (NBT) and forms a chromophore,
diformazan [10]. The superoxide radical scavenging
activity of XG-AgNPs and AO-AgNPs were found to be
dose-dependent (Fig. 7b). The superoxide scavenging
activity XG-AgNPs and AO-AgNPs at 0.1 mg/ml was
18.55 and 17.02% where as at 0.5 mg/ml the activities
were 48.19 and 71.52%. The result of the present study
demonstrated that the IC50 values of XG-AgNPs, AO-
AgNPs and standard antioxidant compound Ascorbic acid
were 0.49, 0.32 and 0.08 mg/ml (Table 1). Hence, AO-
AgNPs showed better superoxide scavenging potential as
compared to XG-AgNPs. The antioxidant potential of
AgNPs could be attributed to functional groups viz. phe-
Fig. 4 FESEM images of silver nanoparticles using the extracts of
a X. granatum and b A. officinalis nolics adhered to them which were originated from the X.
granatum and A. officinalis extracts.

Antioxidant Activity Antidiabetic Activity

The inhibition of carbohydrate metabolizing enzymes like

DPPH is a stable and well-known free radical based on the
a-amylase and a-glucosidase is considered as a useful tool
reduction of accepting hydrogen or electron from donors.
to assess the in vitro antidiabetic potential of different
The DPPH reducing ability of the AgNPs was assessed by
compounds [22]. In the presents study, both XG-AgNPs
observing colour change. When AgNPs added to DPPH
and AO-AgNPs were evaluated for their inhibition poten-
solution, change in colour occurred due to the scavenging
tial against carbohydrate metabolizing enzymes like a-
of DPPH by donation of hydrogen atom to stable the DPPH
amylase and a-glucosidase at three different concentration
molecule [20]. The DPPH free radical scavenging potential
viz. 0.1, 0.2 and 0.5 mg/ml concentrations. The study
of the synthesized XG-AgNPs and AO-AgNPs were eval-
revealed that both XG-AgNPs and AO-AgNPs were able to
uated at 0.1, 0.2 and 0.5 mg/ml and the result is presented
inhibit a-amylase and a-glucosidase enzymes in a dose
in Fig. 7a. The study revealed that both XG-AgNPs and
dependent manner (Fig. 7c, d). The a-amylase inhibition
AO-AgNPs possess free radical scavenging activity in a
potential of XG-AgNPs varied between 35.51 and 89.5%
dose dependent manner. The DPPH scavenging activity
whereas for AO-AgNPs, it ranged between 18.21 and
ranged between 25.6 and 74.85% where as AO-AgNPs
76.13%. From the results of the a-amylase inhibition study,
activity ranged between 31.6 and 100%. The study further
it can be concluded that XG-AgNPs possessed better

123
1110 S. K. Das et al.

Fig. 5 Dynamic Light

Scattering analysis of silver
nanoparticles using the extracts
of a X. granatum and b A.
officinalis

inhibition activity as compared to AO-AgNPs as the former application of external stress or compound, such as strong
is having an IC50 value of 0.19 mg/ml and the latter with acid or base, a concentrated inorganic salt, an organic
0.28 mg/ml. The a-glucosidase inhibition study also solvent or heat. Protein denaturation is a marker for
demonstrated that XG-AgNPs possessed better inhibition inflammatory and arthritic diseases [13]. Most biological
activity as compared to AO-AgNPs as indicated by their proteins lose their biological function when denatured.
IC50 value. The IC50 values of XG-AgNPs and AO-AgNPs Denaturation of proteins (Albumin) is a well-documented
were recorded as of 0.13 and 0.15 mg/ml respectively cause of inflammation. Agents that can prevent protein
(Table 1). Under the similar condition standard antidiabetic denaturation, therefore, would be possible candidate for
drug Acarbose exhibited a-amylase and a-glucosidase anti-inflammatory drug development. As part of the
inhibition activity with IC50 values of 0.15 and 0.11 mg/ml investigation on anti-inflammation activity, the ability of
respectively. The findings of the present study is in nanoparticles to inhibit protein (egg Albumin) denaturation
agreement with earlier study where it has been shown that was evaluated in the present study. Both XG-AgNPs and
mangrove plant extracts mediated silver nanoparticle could AO-AgNPs were evaluated at 0.1, 0.2 and 0.5 mg/ml for
inhibit a-amylase enzyme [14]. their capacity to inhibit the denaturation of heat induced
egg albumin (Fig. 7e). The study demonstrated that both
Anti-inflammatory Activity XG-AgNPs and AO-AgNPs could inhibit the protein
denaturation of albumin in a dose dependent manner and it
Protein (Albumin) denaturation is a process in which pro- ranged between 25.6 and 100%. Amongst the two, AO-
teins lose their tertiary structure and secondary structure by AgNPs showed better protein denaturation capacity with

123
Green Synthesis of Sliver Nanoparticles Using Avicennia officinalis and Xylocarpus granatum… 1111

Fig. 6 XRD analysis of silver a 1800

nanoparticles using the extracts
of a X. granatum and b A. 1600

[4]
officinalis 1400

1200

Intensity (cps)

[1]
1000
800

[2]

[6]
600
400

[5]

[7]
[8]
200

[3]

[10]
[9]
0
10 20 30 40 50 60 70
2-theta (deg)

b 2400
2200
2000
1800

[3]
1600
Intensity (cps)

1400
1200
1000

[6]
[1]
800
600

[2]
400

[7]
[4]

[8]

[10]
200

[9]
[5]
0
10 20 30 40 50 60 70
2-theta (deg)

IC50 value of 0.17 mg/ml. Under the similar condition, the rapidly and exhibit various biological properties and hence
standard drug Asprin could inhibit the denaturation of attracted tremendous attentions of researchers globally.
albumin with IC50 value of 0.41 mg/ml (Table 1). The Different methods are being developed to find new,
present study corroborated with the earlier findings that greener, safer, economical, and rapid synthesis methods for
showed mangrove plant extract mediated AgNPs possesses the preparation of silver nanoparticles. In the present study
anti-inflammatory potential [14]. a simple, safer, greener and non-toxic method of synthe-
Earlier study have shown that different mangrove plants sizing silver nanoparticles has been developed successfully
such as Rhizophora mucronata, Ceriops tagal, Heritiera using X. granatum and A. officinalis plant extracts. The
fomes and Sonneratia apetala, S. alba, S. caseolaris were UV–Vis, FT-IR, DLS and XRD analysis revealed that the
being used for synthesis of silver nanoparticles and their synthesized silver nanoparticles are stable in nature. The
pharmaceutical applications in have also been studied results of the present study also concluded that these syn-
[14, 15, 23, 24]. However the present study on silver thesized silver nanoparticle showed excellent pharmaco-
nanoparticle synthesis using X. granatum and A. officinalis logical properties such as antioxidant, antidiabetic, and
extracts is a new attempt. The therapeutic potential of the anti-inflammatory activities. The synthesized silver
synthesized AgNPs using X. granatum and A. officinalis nanoparticles from the mangrove plant could find potential
plant extracts justified its previously claimed potential applications in biomedical and pharmaceutical industries
applications [25, 26]. and hence, indicate the value of further studies. Thus,
synthesis of silver nanoparticles with medicinal phyto-
chemicals derived from barks of X. granatum and leaves of
Conclusion A. officinalis may result in unprecedented opportunities
directed at large-scale production of silver nanoparticles
Nanotechnology research has grabbed considerable atten- and can be used in many medicinal applications.
tion because of their unique ability to control the matter in
atomic and molecular scale. AgNPs can release of Ag ions

123
1112 S. K. Das et al.

Fig. 7 Bioactivities of silver nanoparticles using the extracts of X. e Protein denaturaion assays. All the experiments were done in
granatum and A. officinalis. a DPPH scavenging; b Superoxide triplicate and data are represented as mean ± SD. Different letters in
scavenging; c a-Amylase inhibition; d a-Glucosidase inhibition; the bar are significantly different (p \ 0.05)

Table 1 Bioactivities (expressed in IC50, mg/ml) of synthesized silver nanoparticles using extracts of X. granatum and A. officinalis
Sample DPPH scavenging Superoxide scavenging a-Amylase inhibition a-Glucosidase inhibition Protein denaturation

XG-AgNP 0.25 ± 0.004 0.49 ± 0.035 0.19 ± 0.011 0.13 ± 0.005 0.27 ± 0.01
AO-AgNP 0.14 ± 0.001 0.32 ± 0.017 0.28 ± 0.002 0.15 ± 0.012 0.21 ± 0.001
Standard 0.04 ± 0.001 0.08 ± 0.002 0.15 ± 0.003 0.11 ± 0.001 0.41 ± 0.01

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Green Synthesis of Sliver Nanoparticles Using Avicennia officinalis and Xylocarpus granatum… 1113

Acknowledgements The authors are thankful to PCCF (Wildlife), 12. E. Apostolidis, Y. I. Kwon, and K. Shetty (2007). Innov. Food
Govt. of Odisha, for giving the necessary permission for the research Sci. Emerg. Technol. 8, 46–54.
work. The authors are also thankful to the DFO, Rajnagar, Odisha and 13. P. Dey, P. Chatterjee, S. Chandra, and S. Bhattacharya (2011). J.
their field staff for their kind help and cooperation during the field Adv. Pharma. Edu. Res. 1, 271–277.
study. The authors would like to acknowledge SAIF, Punjab, India to 14. P. Thatoi, G. K. Rout, S. Gouda, G. Das, K. Pramanik, H.
carry out the 1H, 13C NMR and MS analysis of plant extracts. N. Thatoi, and J. K. Patra (2016). Photochem. Photobiol. 163,
311–318.
15. M. Bakshi, S. Ghosh, and P. Chaudhuri (2015). BioNanoSci. 5,
Compliance with Ethical Standards 162–170.
16. R. Bhanumathi, K. Vimala, K. Shanthi, R. Thangaraj, and S.
Conflict of interest The authors declare that they have no conflict of Kannan (2017). New J. Chem. 41, 14466–14477.
interest. 17. S. Bhakya, S. Muthukrishnan, M. Sukumaran, and M.
Muthukumar (2016). Appl. Nanosci. 6, 755–766.
18. G. Suresh, P. H. Gunasekar, D. Kokila, D. Prabhu, D. Dinesh, N.
References Ravichandran, et al. (2014). Spectrochim. Acta Part A Mol.
Biomol. Spectrosc. 127, 61–66.
1. U. Mani, S. Dhanasingh, R. Arunachalam, E. Paul, P. Shan- 19. M. M. Poojary, P. Passamonti, and A. V. Adhikari (2016). Bio-
mugam, C. Rose, and A. B. Mandal (2013). Prog. Nanotechnol. NanoSci. 6, 110–120.
Nanomater. 2, (1), 21–25. 20. N. Kanipandian, S. Kannan, R. Ramesh, P. Subramanian, and R.
2. M. Rai, K. Kon, N. Duran, S. Galdiero, and M. Galdiero (2014). Thirumurugan (2014). Mater. Res. Bull. 49, 494–502.
Appl. Microbiol. Biotechnol. 98, 1951–1961. 21. P. Nagababu and V. U. Rao (2017). J. Appl. Pharm. Sci. 7,
3. A. Henglein (1998). Chem. Mater. 10, (1), 444–450. 175–182.
4. W. M. Bandaranayake (2002). Wetl. Ecol. Manag. 10, 421–452. 22. H. Ali, P. J. Houghton, and A. Soumyanath (2006). J.
5. M. Khan, M. Khan, S. F. Adil, M. N. Tahir, W. Tremel, H. Ethnopharmacol. 107, 449–455.
Z. Alkhathlan, A. Al-Warthan, and M. R. H. Siddiqui (2013). Int. 23. J. Umashankari, D. Inbakandan, T. T. Ajithkumar, and T. Bala-
J. Nanomed. 8, 1507–1516. subramanian (2012). Saline Syst. 8, 1–7.
6. L. M. Rao and N. Savithramma (2012). Res. Biotechnol. 3, (3), 24. S. P. Dhas, A. Mukerjhee, and N. Chandrasekaran (2013). Int.
41–47. J. Pharma. Pharma. Sci. 5, 349–352.
7. J. Jiang, G. Oberdorster, and P. Biswas (2009). J. Nanopart. Res. 25. H. N. Thatoi, J. K. Patra, and S. K. Das (2014). Acta Physiol.
11, 77–89. Plant 36, 561–579.
8. S. Sun, C. Murray, D. Weller, L. Folks, and A. Moser (2000). 26. H. Thatoi, S. K. Das, and D. Samantaray (2016). Front. Life Sci.
Science. 287, 1989–1992. 9, 267–291.
9. W. Brand-Williams, M. E. Cuvelier, and C. Berset (1995). LWT-.
Food Sci. Technol. 28, (1), 25–30. Publisher’s Note Springer Nature remains neutral with regard to
10. Y. Kono (1978). Arch. Biochem. Biophys. 186, (1), 189–195. jurisdictional claims in published maps and institutional affiliations.
11. F. J. Gella, G. Gubern, R. Vidal, and F. Canalias (1997). Clinica.
Chimica. Acta. 259, 147–160.

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