Hydrangea paniculata flower extract-mediated green synthesis of MgNPs and AgNPs for health care applications

    Hydrangea paniculata flower extract-mediated green synthesis of MgNPs and AgNPs for health care applications Gopalu Karunakaran, Matheswaran Jagathambal, Venkatesh Manickam, Suresh Kumar Govindan, Evgeny Kolesnikov, Arkhipov Dmitry, Alexander Gusev, Denis Kuznetsov PII: DOI: Reference:

S0032-5910(16)30716-1 doi: 10.1016/j.powtec.2016.10.034 PTEC 12033

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Powder Technology

Received date: Revised date: Accepted date:

16 August 2016 30 September 2016 17 October 2016

Please cite this article as: Gopalu Karunakaran, Matheswaran Jagathambal, Venkatesh Manickam, Suresh Kumar Govindan, Evgeny Kolesnikov, Arkhipov Dmitry, Alexander Gusev, Denis Kuznetsov, Hydrangea paniculata flower extract-mediated green synthesis of MgNPs and AgNPs for health care applications, Powder Technology (2016), doi: 10.1016/j.powtec.2016.10.034

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ACCEPTED MANUSCRIPT Hydrangea paniculata flower extract-mediated green synthesis of MgNPs and AgNPs for health care applications

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Gopalu Karunakaran1, 2,*, Matheswaran Jagathambal3, Venkatesh Manickam4, Suresh Kumar

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Govindan4, Evgeny Kolesnikov1, Arkhipov Dmitry1, Alexander Gusev1, 5 and Denis Kuznetsov1 Department of Functional Nanosystems and High-Temperature Materials,

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National University of Science and Technology “MISiS,” Leninskiy Pr. 4, Moscow, 119049, Russia

Department of Biotechnology, K. S. Rangasamy College of Arts and Science,

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Tiruchengode-637215, Tamil Nadu, India Department of Bio-chemistry/Bio-technology/Bio-informatics

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Avinashilingam Institute for Home Science and Higher Education for Women Mettupalayam Road, Bharathi Park Road, Coimbatore -641 043, India Department of Physics, K. S. Rangasamy College of Arts and Science,

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G .R. Derzhavin Tambov State University, 33, Internatsionalnaya Street,

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Tiruchengode-637215, Tamil Nadu, India

Tambov, 392000, Russia

*Corresponding author information Tel: +7-985-663-75-69 E-mail: [email protected], [email protected]

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ACCEPTED MANUSCRIPT Abstract Magnesium nanoparticles (MgNPs) and silver nanoparticles (AgNPs) were synthesized

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by green synthesis method using Hydrangea paniculata flower extract. The occurrence of various organic molecules like terpenoids, steroids, saponins, alkaloids, quinone, glycosides and

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flavonoids in the flower was revealed by phytochemical and gas chromatography–mass

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spectrometry analyses. The formation of brown and black colloids, confirmed the presence of

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MgNPs and AgNPs, which was further supported by ultraviolet-visible absorbance at 360 and 450 nm by MgNPs and AgNPs. The X-ray diffraction analysis confirmed the cubic phase of

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MgNPs and AgNPs. FTIR analysis showed the presence of functional groups on the biosynthesized nanoparticles surface. The SEM and TEM analyses demonstrated the formation

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of spherical and ellipsoidal MgNPs and AgNPs. MgNPs was about 56-107 nm in size, whereas

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AgNPs was approximately 36-75 nm. In addition, both MgNPs and AgNPs showed excellent

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antioxidant potential and antibacterial activity against Escherichia coli and Staphylococcus aureus. The current green synthesis method using H. paniculata flower extract is a simple and an eco-friendly approach to synthesize MgNPs and AgNPs for making beneficial health care products.

Keywords: Green synthesis; Hydrangea paniculata; MgNPs; AgNPs; antioxidant; antibacterial potential.

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ACCEPTED MANUSCRIPT 1 Introduction Green synthesis concept has been booming everywhere since the last few years, and most

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of the researchers are focusing on this area [1]. The results of a recent survey conducted on green

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synthesis showed that it is one of the best and safest methods that can be used for human welfare

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[2]. It involves the synthesis and manipulation of nanoparticles by an eco-friendly approach for making the health care more efficient and effective [3,4].

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Recently, many approaches have been developed under green synthesis for synthesizing

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different nanoparticles. Different green sources such as fungus [5], bacteria [6], algae [7], yeast [8], and plants [9] are used for nanoparticles synthesis. Among these, the use of plant source is

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highlighted, as different parts of plants such as leaf [10], root [11], seed [12], and fruit [13] can

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be used. However, the use of flower extract for the green synthesis of nanoparticles is in the elementary stage. Till date, very few reports are available on flower-mediated green synthesis of

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nanoparticles, compared to other parts of the plants. This kindled our interest to develop a green synthesis approach for nanoparticles synthesis using flower extracts. In a recent research, Allamanda flower extracts have been used for the synthesis of silver nanoparticles (AgNPs) [14]. Hydrangea paniculata is a very common shrub with light pink flower. Its common names are panicled hydrangea and PeeGee hydrangea [15]. This plant is commonly found in various parts of Europe and Asia. The flower is used in the preparation of blood tonic. It contains 4.06% rutin and more than 2.5% phenolic compounds [16]. It also contains different biochemical compounds that are of great importance and medicinal values. It possesses antimalarial, diuretic, and antitussive properties [16]. In recent years, much attention is paid to MgNPs and AgNPs, due to their potential applications in industries, electronics [17], health care (antibacterial, antifungal, anti3

ACCEPTED MANUSCRIPT inflammatory, and wound-healing applications) [18]. Owing to their enormous health care applications, demand for both the nanoparticles has increased. Various approaches are available

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for the synthesis of nanoparticles, in which hazardous chemicals and expensive setup are

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required. Moreover, at the time of nanoparticles recovery, the residue of hazardous chemicals is retained within the nanoparticles, which causes toxic and harmful effects during their

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application. Green synthesis has also received special attention due its inherent advantages such

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as eco-friendly and nontoxic nature, cost effectiveness, simplicity, and easy recoverability. Therefore, the aim of this research was to carry out green synthesis of MgNPs and AgNPs for

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their effective and efficient use in health care applications.

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2 Material and Methods

2.1 Materials Required for Synthesis and Phytochemical Screening

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The precursor material such as magnesium nitrate (Mg(NO3)2) and silver nitrate (Ag(NO3)2) were purchased from Reachem (Russia) and were used for the preparation of the solution for the synthesis. The H. paniculata flowers were collected from Gorky Park (Moscow, Russia). The petals of the flower were collected and washed with sterile water to remove the dust particles and unwanted materials. The washed petals were kept at room temperature in order to drain out the excess water content. Petals (10g) were taken and crushed into an aqueous crude paste using a mortar and pestle, followed by filtration using Whatman No. 1 filter paper. The process of filtration continued until a clear homogeneous extract was obtained. The extract was used to analyze the presence of different phytochemicals such as quinine, flavonoids, glycosides, steroids, alkaloids, saponins, and terpenoids [19]. To confirm the presence of biomolecules, the

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ACCEPTED MANUSCRIPT extract was further characterized by gas chromatography–mass spectrometry (GC-MS)

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(FISONS-GC8000; Canada), with an inbuilt library (NIST and WILEY).

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2.2 Green Synthesis and Structural Characterization of MgNPs and AgNPs Green synthesis of MgNPs and AgNPs were carried out by adding the flower extract in a

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drop-wise manner to 50 ml of 0.2 M solutions each of magnesium nitrate (Mg(NO3)2) and silver

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nitrate (Ag(NO3)2) contained in beakers that were subjected to continuous stirring. The addition of the flower extract was continued until viscous colloids were formed. Thereafter, the solution

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was centrifuged for 25 min at 6300 rpm, until a clear supernatant was obtained, in order to remove the excess extract, non-reactive and other unwanted residues present in the nanoparticle

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suspension. The pellet obtained finally was carefully collected from the bottom of the centrifuge tube, followed by drying at 40-50 °C to remove the excess moisture present in it. The dried pellet

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was crushed into a fine powder using a sterile mortar and pestle. The obtained fine powder was further subjected to various analyses in order to confirm the anticipated nanoparticle formation. The crystalline nature of the synthesized nanoparticles was determined by powder X-ray diffraction (XRD; Difray, Saint Petersburg, Russia) technique using chromium (λ = 2.2909 Å) as X-ray source. Fourier transform infrared (FTIR) spectrophotometer (Nicolet 380; USA) was used to explore the different functional groups present in the nanoparticles. The presence of elements in the nanoparticles was confirmed by energy-dispersive X-ray analysis (EDX SSD; Japan). The morphological features of the synthesized nanoparticles were analyzed by using scanning electron microscope (SEM; JEOL, Japan) and transmission electron microscope (TEM; JEOL). The average size of the synthesized nanoparticles was confirmed by the TEM images. In addition, the pattern obtained by the

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ACCEPTED MANUSCRIPT selected area electron diffraction (SAED) technique was used to correlate the observation with

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XRD results.

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2.3 Antibacterial Susceptibility Test and Antioxidant Potential Analysis The antibacterial susceptibility test for the green synthesized MgNPs and AgNPs was

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performed by disk diffusion protocol developed by Kirby–Bauer [20]. Mueller–Hinton agar

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(MHA) was used to evaluate this test. The organisms used in the test were Escherichia coli (E. coli) (NCIM 2065) and Staphylococcus aureus (S. aureus) (NCIM 2127), which were

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collected from National Collection of Industrial Microorganisms (NCIM, Pune, Maharashtra, India). The test organism broths were prepared 24 h prior to begin the experiment. The turbid

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cultures were aseptically swabbed over the sterile petri plates containing MHA followed by insertion of a 6-mm filter disk (made up of Whatman filter paper) at equal distance in the petri

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plate. After the insertion of the disk, 100 µg/ml nanoparticle suspension was loaded on the filter disk. Similarly, 100 µg/ml streptomycin was used as a positive control. However, an empty disk was used as a negative control and a disk with flower extract was also used. The antioxidant potential of MgNPs and AgNPs was evaluated using the DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging assay [21]. The DPPH solution was freshly prepared and different masses of particles (from 1 to 100 mg) were incubated with the solution. The optical density was measured using an ultraviolet–visible (UV-Vis) spectrophotometer. The obtained results were calculated and tabulated. 3. Results and Discussion The presence of different phytochemicals in the flower extract is shown in Table 1. The majority of organic compounds were found to be terpenoids, steroid, saponins, alkaloids,

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ACCEPTED MANUSCRIPT quinone, glycosides and flavonoid. GC–MS results showed the presence of different specific organic molecules and are shown in Figure 1 and Table 2. The obtained GC–MS chromatograph

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showed sharp peaks at different retention times, which is specific for different compounds. The

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majority of the organic compounds are shown in Table 2. The obtained results clearly showed that these compounds play a significant role as reducing and stabilizing agents during the

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synthesis.

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During the green synthesis, the formation of nanoparticles was observed by change in the color of the solution. The results are shown in Figure 1. Mg(NO3)2 solution was colorless. When

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it was mixed with flower extract, it turned light brown and finally to muddy brown, indicating MgNP formation, as shown by a recent research on Emblica officinalis extract-mediated

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synthesis of MgNPs [22]. Similarly, when the colorless Ag(NO3)2 solution was mixed with flower extract, its color changed to light brown and then to dark brown, confirming the formation

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of AgNPs. Similar kind of color changes was observed, when AgNPs were synthesized using the extracts of Terminalia chebula [23]. The maximum absorption peak for MgNPs was obtained at 360 nm under UV–Vis spectroscopy, which is shown in Figure 2a. The XRD result for MgNPs is shown in Figure 2b. The obtained 2θ values were found to be in line with the ICDD (International Centre for Diffraction Data) standard data files (JCPDS: 00-001-1235) for magnesium oxide (MgO), confirming the presence of a cubic crystal system in the synthesized MgNPs. The pattern exhibits reflection from (111), (200), (220), and (311) Miller’s plane at ~55.6°, ~65.7°, ~100.1° and ~128.9°, respectively. In addition, a broad hump was observed at ~20.9°, which was due to the presence of some organic molecules in the nanoparticles [23].

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ACCEPTED MANUSCRIPT The absorption peak for AgNPs was observed at 460 nm under UV–Vis spectroscopy, which is shown in Figure 3a. Figure 3b shows the XRD results of AgNPs. The obtained 2θ

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values were found to be in line with the ICDD standard data files (JCPDS: 00-001-1167) for silver (Ag), confirming the presence of a cubic crystal system in the synthesized AgNPs. The

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pattern exhibited reflection from (111), (200), (220), and (311) Miller’s plane at ~58.1°, ~68.35°,

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~105.0°, and ~136.7°, respectively. However, the broad humps observed at ~47.5° and ~93.5°

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are due to the presence of organic molecules and oxygen present in the synthesized nanoparticles [24].

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The obtained FTIR result for MgNPs is shown in Figure 2c. Sharp peaks were observed at 3378, 2627, 2290, 2049, 1406, and 866 cm1. The bands observed at 1406 and 866 cm1

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represented the presence of MgNPs. Those observed at 3378 and 2627 cm1 corresponded to the –OH functional group, which may be due to the presence of organic molecules such as ester,

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phenols, and acids [25]. The prominent bands observed at 2290 and 2049 cm1 correspond to different functional groups such as –C–H, –CH, and C=O in the nanoparticles, which may be due to the presence of aldehyde and amides [25]. Similarly, FTIR result for AgNPs is shown in Figure 3c. Different peaks were observed at 3658, 2494, 2220, 1978, 1368, 1247, 1038, and 872 cm1. The bands observed at 1038 and 872 cm1 confirmed the presence of AgNPs. Those observed at 3658 and 2494 cm1 corresponded to the O–H group, which is due to the presence of organic molecules such as phenols, esters, and carboxylic acid in the NPs. Similarly, the bands observed at 2220, 1978, 1368, and 1247 cm1 corresponded to the presence of different functional groups such as –C–H, C=O, and –CH [25]. These results clearly showed that these different functional groups may participate in the reduction and stabilizing reactions during nanoparticle formation. 8

ACCEPTED MANUSCRIPT The EDX analysis confirmed the presence of elements in the synthesized MgNPs and AgNPs. The observed results showed the counts due to X-ray on the vertical axis and the keV

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values on the horizontal axis. Thus, on the basis of the keV values, the interpretation is made for

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the characterization of the biosynthesized nanoparticles. Figure 2d shows the EDX analysis of MgNPs, confirming the presence of Mg, carbon, and oxygen in the synthesized MgNPs.

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Similarly, Figure 3d represents the EDX analysis of AgNPs, confirming the presence of Ag,

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carbon, and oxygen in the synthesized AgNPs. The presence of carbon could be due to involvement of organic molecules during the reduction and stabilizing process of nanoparticle

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formation. Similarly, the presence of oxygen may be due to the formation of an intermediate complex or a stable molecule during the reaction mechanism of nanoparticle formation.

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The morphological characteristics of the green synthesized MgNPs and AgNPs were determined using SEM and TEM analyses. Figure 2e and 2f shows the SEM and TEM images of

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MgNPs. These images revealed that most of the particles are spherical and ellipsoidal in nature. In addition, the particle size was found to be in the range of 56 to 107 nm, with an average particle size of 87±4 nm for MgNPs. Similarly, Figure 3e and 3f shows the SEM and TEM images of AgNPs. These images clearly showed the spherical, oval, and ellipsoidal nature of the biosynthesized AgNPs. The particle sizes for the AgNPs were found to be in the range from 36 to 75 nm, with an average particle size of 55±5 nm. The SAED patterns of the synthesized MgNPs and AgNPs are shown in Figures 2f and 3f. These patterns were found to be well in agreement with those observed from the XRD peaks and demonstrated the crystalline phase. The radical scavenging potential of the synthesized MgNPs and AgNPs was analyzed using the DPPH method, and the observed results are shown in Figure 4a and 4b. Figure 4a showed the radical scavenging activity of the MgNPs treated at different concentrations: 12%

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ACCEPTED MANUSCRIPT (1 mg), 28% (5 mg), 48% (10 mg), and 78% (100 mg). Similarly, the radical scavenging activity of the AgNPs treated at different concentrations was observed to be 14% (1 mg), 32% (5 mg),

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55% (10 mg), and 86% (100 mg). The AgNPs synthesized using extract of Citrullus lanatus [26]

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and Clerodendrum phlomidis [27] extracts exhibited good antioxidant activity. Both the studies showed a concentration-dependent radical scavenging activity of AgNPs.

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The antibacterial potential was evaluated using Kirby–Bauer method. The results are

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shown in Table 3 and Figure 4c and 4d. Antibiotic streptomycin was used as the standard drug (positive control) for comparison. The antibacterial activities of the nanoparticles against E. coli

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(Gram negative) and S. aureus (Gram positive) were tested. From the observed results, it was clear that MgNPs exhibited 13±0.21 and 13±0.24 mm zone of inhibition against E. coli and

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S. aureus, respectively. Similarly, AgNPs showed 14±0.43 and 13±0.14 mm zone of inhibition against E. coli and S. aureus, respectively. The biosynthesized nanoparticles exhibited a greater

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antibacterial activity than the standard antibiotic (streptomycin) against the tested bacterial strains. However, MgNPs showed lesser antibacterial activity than AgNPs. The variation in the inhibition zone was also noticed with respect to the type of bacterium, which might be due to the difference in the bacterial surface characteristics. The Gram-positive bacterium possesses a thick peptidoglycan layer, whereas the Gram-negative bacterium possesses a thin peptidoglycan layer. The NPs could easily penetrate the thin layer of Gram-negative bacterium and kill them more effectively, when compared to Gram-positive bacterium through electron-mediated mechanisms [28, 29]. Another reason for the antibacterial reactivity of nanoparticles may be the generation of reactive oxygen species or inactivation of enzymes resulting in cell death [30]. The results of this study clearly demonstrated that the biosynthesized nanoparticles exhibited antibacterial activity, by inhibiting the growth of harmful bacteria through different mechanisms.

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ACCEPTED MANUSCRIPT 4 Conclusions Green synthesis of MgNPs and AgNPs using the extract of H. paniculata flower is one of

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the non-toxic, effective, economical and safe approaches. The biomolecules found in flower

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played an important role during MgNP and AgNP synthesis under normal temperature conditions. The XRD analysis confirmed the presence of cubic crystalline phase in both the

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synthesized nanoparticles. The FTIR analysis confirmed the involvement of organic molecules

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during green synthesis. The EDX analysis confirmed the presence of carbon and oxygen during nanoparticles formation. The microscopic images obtained using SEM and TEM techniques

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showed that the green synthesized particles are spherical, ellipsoidal, and oval, with an average size of about 87±4 and 55±5 nm for MgNPs and AgNPs, respectively. MgNPs and AgNPs tested

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against E. coli and S. aureus, confirmed their remarkable antibacterial activity. Also, the DPPH method showed that the green synthesized nanoparticles have tremendous radical scavenging

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activity. Therefore, this green synthesis is a prospective approach for preparing MgNPs and AgNPs for many health-care applications.

Acknowledgement

The authors gratefully acknowledge the financial support of the Ministry of Education and Science of the Russian Federation in the framework of Increase Competitiveness Program of NUST «MISiS» (№ К4-2015-017). References 1. A. Kanchana, S. Devarajan, S. R. Ayyappan, Green synthesis and characterization of palladium nanoparticles and its conjugates from solanum trilobatum leaf extract, Nano-Micro Lett. 2, (2010) 169-176.

11

ACCEPTED MANUSCRIPT 2. S. Ahmed, M. Ahmad, B. L. Swami, S. Ikram, A review on plants extract mediated synthesis of silver nanoparticles for antimicrobial applications: A green expertise. J. Adv. Res. 7 (1),

PT

(2016) 17-28.

RI

3. V. Ravichandran, S. Vasanthi, S. Shalini, S.A.A. Shah, R. Harish, Green synthesis of silver nanoparticles using Atrocarpus altilis leaf extract and the study of their antimicrobial and

SC

antioxidant activity. Mater. Lett. 180, (2016) 264-267.

Powder Technol. 294, (2016) 280-283.

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4. T. Wang, S. Liu, Green synthesis and photoluminescence property of AlOOH nanoflakes,

MA

5. B.S. Fathima, R.M. Balakrishnan, Biosynthesis and optimization of silver nanoparticles by endophytic fungus Fusarium solani. Mater. Lett. 132, (2014) 428-431.

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6. N. Srivastava, M. Mukhopadhyay, Biosynthesis and structural characterization of selenium nanoparticles mediated by Zooglea ramigera, Powder Technol. 244, (2013) 26-29.

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7. Z. Salari, F. Danafar, S. Dabaghi, S. A. Ataei, Sustainable synthesis of silver nanoparticles using macroalgae Spirogyra varians and analysis of their antibacterial activity. J. Saudi Chem. Soc. 20 (4), (2016) 459-464. 8. G. Xu, X. Zhang, H. Cui, Z. Zhang, J. Ding, J. Wu, Facile synthesis of mesoporous SnO 2 microspheres using bioactive yeast cell, Powder Technol. 301, (2016) 96-101. 9. S.C.G.K. Daniel, L.A.N. Julius, S.S. Gorthi, Instantaneous detection of melamine by interference biosynthesis of silver nanoparticles. Sens. Actuators B Chem. 238, (2017) 641650. 10. J. L. Lopez-Miranda, M. Vazquez, N. Fletes, R. Esparza, G. Rosas, Biosynthesis of silver nanoparticles using a Tamarix gallica leaf extract and their antibacterial activity. Mater. Lett. 176, (2016) 285-289.

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ACCEPTED MANUSCRIPT 11. C. Sudhakar, K. Selvam, M. Govarthanan, B. Senthilkumar, A. Sengottaiyan, M. Stalin, T. Selvankumar, Acorus calamus rhizome extract mediated biosynthesis of silver nanoparticles

PT

and their bactericidal activity against human pathogens. J. Genet. Eng. Biotechnol. 13(2),

RI

(2015) 93-99.

12. C. Luna, V.H.G. Chavez, E.D. Barriga-Castro, N.O. Nunez, R. Mendoza-Resendez,

SC

Biosynthesis of silver fine particles and particles decorated with nanoparticles using the

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extract of Illicium verum (star anise) seeds. Spectrochim. Acta A Mol. Biomol. Spectrosc. 141, (2015) 43-50.

MA

13. B. Moldovan, L. David, M. Achim, S. Clichici, G.A. Filip, A green approach to phytomediated synthesis of silver nanoparticles using Sambucus nigra L. fruits extract and

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D

their antioxidant activity. J. Mol. Liq. 221, (2016) 271-278. 14. G. Karunakaran, M. Jagathambal, A. Gusev, E. Kolesnikov, A.R. Mandal, D. Kuznetsov

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(2016). Allamanda cathartica flower's aqueous extract-mediated green synthesis of silver nanoparticles with excellent antioxidant and antibacterial potential for biomedical application. MRS Commun. 6 (01), (2016) 41-46. 15. D. Michael, Hydrangeas for American Gardens. Timber Press, Portland, OR, ISBN 0-88192641-8 (2004).

16. C. J. van Gelderen, D. M. van Gelderen, Encyclopedia of Hydrangeas. Timber Press, Portland, OR, ISBN 0-88192-622-1 (2004). 17. S.K. Moorthy, C.H. Ashok, K. V. Rao, C. Viswanathan, Synthesis and Characterization of Mgo Nanoparticles by Neem Leaves through Green Method. Materials Today: Proceedings 2(9), (2015) 4360-4368.

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ACCEPTED MANUSCRIPT 18. B. Ajitha, Y. Ashok Kumar Reddy, P. Sreedhara Reddy, Enhanced antimicrobial activity of silver nanoparticles with controlled particle size by pH variation, Powder Technol. 269,

PT

(2015) 110-117.

RI

19. A.M. Ismail, E.A. Mohamed, M.R. Marghany, F.F. Abdel-Motaal, I.B. Abdel-Farid, M.A. El-Sayed, Preliminary phytochemical screening, plant growth inhibition and antimicrobial

SC

activity studies of Faidherbia albida legume extracts. J. Saudi. Soc. Agr. Sci. (2014) doi:

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10.1016/j.jssas.2014.06. 002

20. A.W. Bauer, W.M. Kirby, J.C. Sherris, M. Turck, Antibiotic susceptibility testing by a

MA

standardized single disk method. Am. J. Clin. Pathol. 45, (1966) 493-496. 21. A. Serpen, E. Capuano, V. Fogliano, V. Gokmen, A New Procedure to Measure the

7681.

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Antioxidant Activity of Insoluble Food Components. J. Agric. Food Chem. 55, (2007) 7676-

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22. K. Ramanujam, M. Sundrarajan, Antibacterial effects of biosynthesized MgO nanoparticles using ethanolic fruit extract of Emblica officinalis. J. Photochem. Photobiol. B 141, (2014) 296-300.

23. C.S. Espenti, K.S.V.K. Rao, K.M. Rao, Bio-synthesis and characterization of silver nanoparticles using Terminalia chebula leaf extract and evaluation of its antimicrobial potential. Mater. Lett. 174, (2016) 129-133. 24. M. Govindarajan, M. Rajeswary, U. Muthukumaran, S.L. Hoti, H.F. Khater, G. Benelli, Single-step biosynthesis and characterization of silver nanoparticles using Zornia diphylla leaves: A potent eco-friendly tool against malaria and arbovirus vectors. J. Photochem. Photobiol. B 161, (2016) 482-489.

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ACCEPTED MANUSCRIPT 25. C. Dipankar, S. Murugan, The green synthesis, characterization and evaluation of the biological activities of silver nanoparticles synthesized from Iresine herbstii leaf aqueous

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extracts. Colloids Surf. B 98, (2012) 112–119.

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26. J.K. Patra, G. Das, K.H. Baek, Phyto-mediated biosynthesis of silver nanoparticles using the rind extract of watermelon (Citrullus lanatus) under photo-catalyzed condition and

SC

investigation of its antibacterial, anticandidal and antioxidant efficacy. J. Photochem.

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Photobiol. B 161, (2016) 200-210.

27. R. Sriranjani, B. Srinithya, V. Vellingiri, P. Brindha, S.P. Anthony, A. Sivasubramanian,

MA

M.S. Muthuraman, Silver nanoparticle synthesis using Clerodendrum phlomidis leaf extract and preliminary investigation of its antioxidant and anticancer activities. J. Mol. Liq. 220,

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(2016) 926-930.

28. Q.L. Feng, J. Wu, G.Q. Chen, F.Z. Cui, T.N. Kim, J.O. Kim, A mechanistic study of the

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antibacterial effect of silver ions on Escherichia coli and Staphylococcus aureus. J. Biomed. Mater. Res. 52(4), (2000) 662-668. 29. S. Rajeshkumar, C. Malarkodi, M. Vanaja, G. Annadurai, Anticancer and enhanced antimicrobial activity of biosynthesizd silver nanoparticles against clinical pathogens. J. Mol. Struct. 1116, (2016) 165-173.

30. D. Manikprabhu, J. Cheng, W. Chen, A.K. Sunkara, S.B. Mane, R.Kumar, M. das, W.N. Hozzein, Y.Q. Duan, W.J. Li, Sunlight mediated synthesis of silver nanoparticles by a novel actinobacterium (Sinomonas mesophila MPKL 26) and its antimicrobial activity against multi drug resistant Staphylococcus aureus. J. Photochem. Photobiol. B 158, (2016) 202-205.

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ACCEPTED MANUSCRIPT Figure captions Figure 1. Green synthesis of MgNPs and AgNPs: Schematic representation of phytochemicals

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involvement in the biosynthesis mechanism of nanoparticles.

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Figure 2. Characterization of MgNPs a) UV-Vis b) XRD c) FTIR spectrum d) Elemental analysis (EDS) e) SEM, f) TEM and SAED pattern.

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Figure 3. Characterization of AgNPs a) UV-Vis b) XRD c) FTIR spectrum d) Elemental analysis

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(EDS) e) SEM, f) TEM and SAED pattern.

Figure 4. a and b) Antioxidant activity of MgNPs and AgNPs (*, ** Represents level of significant

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at p<0.05) and c and d) antibacterial activity of MgNPs and AgNPs (A: 100 µg/mL antibiotic, B: Flower extracts, C: only disk, D: 100 µg/mL AgNPs, E: 100 µg/mL MgNPs and F: distilled

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water).

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ACCEPTED MANUSCRIPT Table 1. Phytochemical analysis of Hydrangea paniculata flower extracts

Steroid

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Saponins

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Alkaloids

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Quinone

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Glycosides

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Flavonoid

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Terpenoids

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Results

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Phytochemicals Name

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S. No.

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+ Indicates the present of compound flower extracts

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+ + + +

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S. No.

Retention

Compounds Name*

Time 1.

4.61

Methyl 6-hydroxy-3,5-

5-(2-Methoxyethyl)-3-phenylisoxazole

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15.64

1-Hexadecene

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20.18

Phytol acetate

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23.03

Eicosyl acetate

6.

25.26

Octadecane

7.

29.38

Bis-(3,5,5-trimethylhexyl) ether

8.

32.00

1,2-Benzenedicarboxylic acid, diisooctyl

NU

MA

1,2-Diphenyl-1,2-dithiocyanatoethane

D

35.19

formula

Weight

(%)

174

0.80

C12H13NO2

203

0.66

C16H32

224

1.16

C22H42O2

338

5.27

C22H44O2

340

1.63

C18H38

254

36.45

C18H38O

270

0.99

C24H38O4

390

31.23

C16H12N2S2

296

0.64

RI

10.95

SC

2.

9.

Molecular Area

C9H18O3

dimethylhexanoate

ester

Molecular

PT

Table 2. GC-MS analysis of Hydrangea paniculata flower extracts

AC CE P

TE

*Compounds where analyzed by comparing the peaks with the library (NIST and WILEY)

22

ACCEPTED MANUSCRIPT

Table 3. Antibacterial activity of MgNPs and AgNPs nanoparticles

PT

Zone of inhibition

Microorganisms

Streptomycin

10±0.21

Staphylococcus aureus

11±0.13

AC CE P

TE

D

MA

NU

Escherichia coli

23

MgNPs

AgNPs

(100 µg/ml)

(100 µg/ml)

13±0.28

14±0.43

13±0.24

13±0.14

SC

(100 µg/ml)

RI

[Mean + SD (mm)]

ACCEPTED MANUSCRIPT

AC CE P

TE

D

MA

NU

SC

RI

PT

Graphical Abstract

24

ACCEPTED MANUSCRIPT Research Highlights

Green synthesis of MgNPs and AgNPs using Hydrangea paniculata flower extract



Phytochemical screening of flower extract by biochemical and GC-MS analyses



Structural characterization of synthesized nanoparticles



Antioxidant and Antibacterial activities of synthesized nanoparticles

AC CE P

TE

D

MA

NU

SC

RI

PT



25