Biosynthesis of silver and gold nanoparticles using Musa acuminata colla flower and its pharmaceutical activity against bacteria and anticancer efficacy

Journal Pre-proof Biosynthesis of silver and gold nanoparticles using Musa acuminata colla flower and its pharmaceutical activity against bacteria and anticancer efficacy

Saritha Valsalam, Paul Agastian, Galal Ali Esmail, Abdul-Kareem Mohammed Ghilan, Naif Abdullah Al-Dhabi, Mariadhas Valan Arasu PII:

S1011-1344(19)31293-X

DOI:

https://doi.org/10.1016/j.jphotobiol.2019.111670

Reference:

JPB 111670

To appear in:

Journal of Photochemistry & Photobiology, B: Biology

Received date:

26 September 2019

Revised date:

20 October 2019

Accepted date:

23 October 2019

Please cite this article as: S. Valsalam, P. Agastian, G.A. Esmail, et al., Biosynthesis of silver and gold nanoparticles using Musa acuminata colla flower and its pharmaceutical activity against bacteria and anticancer efficacy, Journal of Photochemistry & Photobiology, B: Biology(2019), https://doi.org/10.1016/j.jphotobiol.2019.111670

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© 2019 Published by Elsevier.

Journal Pre-proof

Biosynthesis of Silver and Gold nanoparticles using Musa acuminata colla flower and its pharmaceutical activity against bacteria and anticancer efficacy

Saritha Valsalam1,, Paul Agastian1,*, Galal Ali Esmail2, Abdul-Kareem Mohammed Ghilan2, Naif Abdullah Al-Dhabi2, Mariadhas Valan Arasu2 1

Department of Plant Biology & Biotechnology, Loyola College (Affiliated to Madras

Department of Botany and Microbiology, College of Science, King Saud University, P. O.

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Box 2455, Riyadh 11451, Kingdom of Saudi Arabia

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2

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University), Chennai 600 034, Tamil Nadu, India

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[email protected]

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*Corresponding Author

Journal Pre-proof Abstract Synthesis of nanoparticles using plant sources as reducing agent has become important, as physical and chemical methods are costlier and affects environment. Hence it is important to develop environment friendly nanoparticle synthesis by avoiding the use of toxic chemicals. The present study aimed to synthesize silver nanoparticles (Ag Nps) and gold nanoparticles (AuNps) using Musa acuminata colla flower and its pharmaceutical activity against extended spectrum beta-lactamase (ESBL) gene producing bacteria and anticancer efficacy. The synthesized Ag and Au NPs were analysed by means of UV-Vis, FTIR, XRD,SEM and

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EDAX evidenced the bioreduction of Ag+ ions to Ag0 and Au3+ ions to Au0 respectively. Both nanoparticles and flower extracts were studied for antibacterial activity of ESBL gene

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producing bacteria by disc diffusion and microdilution (Resazurin) method. In vitro anticancer efficacy (MCF-7) and toxicity (VERO) of AgNPs, AuNPs, aqueous extract and

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ethanol extract of flowers were performed by MTT assay. IC50 value for DPPH analysis was

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at 390µg and 460µg for ethanol and aqueous extract respectively. Total antioxidant content was found be 740 µg/mg and 460µg/mg for ethanol and aqueous extract. GCMS analysis

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authenticated the existence of the compounds namely, 9,12-octadecadienoic acid(z,z)- and nhexadecanoic acid in the crude extract of the samples. Among the samples, AgNPs had best

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antibacterial activity. AgNPs and AuNPs were confirmed by colour change to reddish brown and ruby red. Further ƛmax were obtained at 474 and 540 nm by UV - visible spectrum. SEM

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analysis revealed the particle size ranges from 12.6 to 15.7 nm for silver and 10.1 to 15.6 nm for gold nanoparticles. The EDAX spectrum shows a strong signal for elemental Ag and Au at ~ 3 keV and 1.5 keV. The XRD patterns for silver and gold nanoparticles at 36.701, 42.900, 63.281 and 76.398 corresponding to the lattice planes 2.4467, 2.1064, 1.46839, 1.24564 nm and 27.32, 36.7228, 39.56, 42.888, 63.253, 63.253, 65.02 and 76.383 corresponding to the lattice planes 3.262, 2.44530, 2.276, 2.1070, 1.46897, 1.4332 and 1.24585 nm. The IC50 values for MCF-7 and VERO cells were 30.0 µg/ml and 55.0µg/ml respectively. Key words: Musa acuminata colla, Silver, Gold nanoparticles, UV - visible spectrum, FTIR, XRD, GC-MS, Anticancer

Journal Pre-proof Introduction Synthesis of nanoparticles from plants and its sources is currently an emerging field of nanobiotechnology. Size of nanoparticles (NP) ranges from 1- 100nm. Specific size, shape, morphology, physical, chemical nature and alignment have enhanced its property. NP are used in manufacturing shampoos, soaps, detergents, cosmetics, toothpastes and shoes. In addition, NP have major application in medical and therapeutic products. Silver and silver nitrate possess antimicrobial activity there by it is used in synthesis of plant based NP for reduction [1]. Generally, -metal NP such as gold, silver and platinum metals have been used

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as green method for the biosynthesis of NP [2]. NP synthesized using the biological sample by an eco-friendly method has wide level of biological applications such as antimicrobial,

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anticancer, antidiabetic and immunomodulatory and liver associated diseases and disorders [2].

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In the field of biotechnology, biosynthesized NP has become importance due to its

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effectiveness in synthesis of metal based green NP. Non-toxic silver has capacity to fight against numerous microbes. Physical and chemically synthesised NP may contain small

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amount of unreacted particles which will lead to side effects. Green synthesised NP has distinctive electrical, optical and biological properties. Biosynthesis of NP is a single step

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process without cost effective and not tedious process. This makes them suitable catalyst, biosensor, drug delivery and fabrication [3]. Advantages of biological method in NP

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synthesis are controlled crystal growth, stability and faster synthesis [4]. Banana is rich in flavonoids, polyphenols, anthocyanins, minerals, bioactive amines like dopamines, serotonin which could be the possible reducing and capping agent for NP biosynthesis [5]. Banana peel is used as shoe polish, teeth whitener and pain reliever. It is also used to treat bites by bugs, psoriasis and to reduce acuteness of the arthritis and aches [6]. Antioxidant content of the plants play major role for its biological activity such as intervening in different phases of cancer development involving initiation, progression, promotion, invasion and metastasis [7]. Nowadays biological methods are preferred because of its non-toxicity and less cost. Musa barbisiana is rich in polyphenol, carotenoids, dietary fibre, proteins, vitamins, energy, minerals, unsaturated fatty acids and potassium [8]. The atomic, molecular and supra-molecular molecules of nanotechnology synthesised particles improves its functionalities [9].Cancer is a disease caused due to uncontrolled cell growth, later stage of which is referred as metastasis. Even though there is lot of development in medicinal field lack of diagnosis in early stage makes them untreatable. The disadvantage of

Journal Pre-proof chemotherapy is overcome by alternate treatment such as plant based drugs as they possess natural anti-angiogenic effect [10]. The pulp and peel of banana contains many phytonutrients. Therefore, they are rich source of energy and nutrition. High availability and easy consumption makes them source of edible vaccine. It is used to treat degenerative diseases. The pectin isolated from banana is used in tablet formulation. Additive and synergistic effects of fruits and vegetables increase the potency against cancer [11].Hence the present study aims to utilize the above constituents of banana for the biosynthesis if NP using silver and gold and studied for its ailments against

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ESBL gene producing bacteria and cancer cell lines.

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2. Materials and Methods

Silver

nitrate

(AgNO3,

99.9

%)

and

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2.1. Chemicals and reagents

hydrogen

tetrachloroaurate(III)

trihydrate

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(HAuCl4·3H2O, 99.9 %) was acquired from Sigma-Aldrich, USA. Different cultivation

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medium such as nutrient agar and glucose were purchased from Merc, India. The routine laboratory, chemicals and solvents were obtained from the local vendors of Chennai, India.

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2.2. Plant Materials

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The standard antibiotic discs have been obtained from Himedia.

The flowers of the Musa acuminata colla were collected from Nagercoil, Kannya Kumari, India. Subsequently, the flowers were subjected to washing process with distilled water to eliminate the dust particles, shade dried and powdered for further research work.

2.3. Soxhlet extracts procedure for plants Ethanol and aqueous extracts were extracted by means of standard approaches of Indian Pharmacopoeia. The dried leaves (~25 g) were kept in a Soxhlet method with the support of organic solvent ethanol, which was then heated to reflux. 2.4. Phytochemical screening Typically, in qualitative analysis, the obtained filtrate was verified to determine the presence of phytochemicals using standard procedures.

Journal Pre-proof 2.5. Antioxidant activity DPPH and total antioxidant assays were performed to determine the antioxidant content of flower extract [12, 13].

2.6. BioSynthesis of Ag and AuNPs In a typical process, 10 ml aqueous extract of M. acuminata colla flowers were added to 90 ml of 1.0 mM aq. AgNO3 and HAuCl4·3H2O solution at room temperature to synthesis Ag and Au NPs respectively. In particular, there was a visible-color change from reddish-brown

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and ruby-red, which was monitored with UV-Vis spectrophotometer. Further, the changes in colours confirms the bio-reduction of Ag+/Ag0 and Au3+/Au+ for plant-extract-mediated

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synthesis [14]. Lastly, Ag and Au NPs were acquired by centrifugation process at 10,000 rpm

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for 30 min. Then it was subjected to washing process with distilled water thrice and air dried.

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The obtained powder was kept in the refrigerator for instrumental and activity analysis. 2.7. Characterization of Ag and Au NPs

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The spectral nature of Ag and Au NPs was performed on Thermo Fisher UV–Vis spectrophotometers. FT-IR spectra of fabricated Ag and Au NPs were recorded in the range

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of 4000–400 cm−1 by means of Jasco 6300 spectrometers. The morphological changes of the synthesised NPs was assessed using SEM (TESCAN VEGA3 SBU) armed with EDAX. The

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crystalline properties of Ag and Au NP were performed on a X-ray diffractometer–Cu Kα radiation system (Rigaku-Miniflex-600, Japan). 2.8. Antibacterial Activity

The pure culture of ESBL gene producing bacteria (Bacterial cultures - Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, Proteus mirabilis and Klebsiella pneumoniae) were isolated from food samples and tested for antibacterial measurements. Antibacterial behaviour of Soxhlet extracted samples, Ag and Au NP were assessed by agar disc diffusion and broth dilution (resazurin) method [15]. 2.9. In vitro cytotoxicity activity Cell lines were procured from NCCS Pune. The cells were placed in Minimal Essential Media accompanied with 10% FBS and antibiotics. The in vitro anticancer behaviours of Ag and AuNP on MCF-7 and normal Vero cells were evaluated by the MTT assay [16].

Journal Pre-proof Subsequently, the assay was performed through various concentrations of Ag and AuNP to find out 50% of cell viability, assessed via the below equation: % cell viability = [absorbance of treated cells at 540 nm / absorbance of treated cell at 540 nm] × 100 2.10. Phytochemical components identification by GCMS The metabolite profiling of the plant extracts were identified by GCMS analysis by following

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the methodology of Sumathy et al., 2011 [17].

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3. Results and Discussion

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3.1 Extraction of flower

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The M. acuminata colla flowers were shade dried and extracted by means of ethanol and aqueous. The obtained yields were assessed in grams and the maximal yield was largely

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3.2 Phytochemical evaluation

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obtained in ethanol extract (2.32g/25g) followed by aqueous extract (1.12g/25g)

Phyto-constituents of the extracts were analyzed qualitatively. In both extracts proteins,

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alkaloids, steroids and cardio-glycosides were present. In addition, ethanol extract contains tannins, flavanoids and anthro-quinone (Table 1). These secondary metabolites might be responsible for therapeutic properties. Similarly, Sumathy et al., 2011 [17] reported the presence of glycosides, tannins, saponins, steroids, phenols and flavonoids in methanol extract of Musa acuminata colla flower. Obiageli et al., 2016 [18] reported Musa species revealed the existence of alkaloids, saponins, glycosides and flavonoid different stages (ripe and unripe). Tannin was found only in the ripe stage of banana Al-Dhabi et al., 2019 [19] .

3.3 Antioxidant activity For DPPH assay ethanol extract displayed IC50 value at 390 µg and aqueous extract 460 µg whereas BHT showed 170 µg. Total antioxidant activity showed 460µg/ml for aqueous and 740µg/ml for ethanol extracts (Fig.1). Dahham et al., 2015 [11] reported ethanolic extract of M. sapientum fruit exhibited IC50 value at 19.10 μg/mL for DPPH scavenging analysis. Imam

Journal Pre-proof et al ., 2011 [20] reported total antioxidant capacities of M. sapientum ssp. sylvestris peel, pulp and seed extract 675.56 ± 5.23, 543.70 ± 3.14 and 416.85 ± 5.50 mg/g as IC50 values.

3.4 Synthesis of Ag and Au NPs The changes in colour to reddish brown specifies the bioreduction of Ag ions to Ag0 and to colour of ruby red shows the reduction of Au3+ to Au0. The observed colour changes suggest the conformation of Ag and AuNPs (Fig. 2).

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3.5 UV–visible spectroscopy

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The fabricated Ag and Au NPs synthesized using M. acuminata colla flowers were analysed by UV–Vis absorption spectra, where SPR band was detected at 540 nm for Au NPs and for

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474 nm for Ag NPs and the results are displayed in Fig. 3. Mukundan et al., 2015 [21] reported that SPR band for Ag and Au NPs was at 441 nm and 563 nm using Bauhinia

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3.6 FTIR analysis

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tomentosa leaves.

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FT-IR spectra of fabricated samples were logged to determine the probable biomolecules accountable for the bio-reduction of metal ions and capping the NP. The obtained Ag and Au

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nano solutions were centrifuged at 10000 rpm for 30 min and air-dried. FT-IR spectrum of synthesised AgNP and Au NPs from M. acuminata colla flowers is displayed in Fig. 4. The flower extract possesses distinctive vibrational peaks at 3418, 2923, 2139, 1639, 1424, 1322, 1248, 1054 and 621cm−1. Distinctive vibrations after the reduction of Ag+ and Au3+ ions by M. acuminata colla flower extract were found to be 3427, 2924, 1626, 1383, 1321, 1112, 791, 764, 712, 564 cm−1 and 3421, 2924, 2855, 2357, 1642, 1549, 1404, 1072 and 598 cm−1 correspondingly. Particularly, the Ag NPs have peaks at 1112, 1383, and 1626 cm−1, whereas Au NPs reveals peaks at 1072, 1404, and 1642 cm−1. The observed functional groups have a major part in stability/capping of fabricated nanoparticles as described in the literature [22]. The observed bands at 1383 cm−1 and 1112 cm−1 were related to the N–H and C–N (amines) stretch vibration of the proteins correspondingly. Thus the observed major peaks present in the flower extract are also present in both Ag and Au NPs concluding that the existence of phytoconstituents which are exist in the fabricated NPs. The existence of several functional

Journal Pre-proof groups accountable for the synthesis of AgNPs synthesized using neem (A. indica) leaf broth [22].

3.7 SEM - EDAX analysis 3.7.1 SEM analysis SEM analysis of flower extracted Ag and AuNPs are displayed (Fig. 5). Ag NPs revealed the existence of wide range of poly-dispersed spherical shaped particles and tends to have size ranging from 12.6 to 15.7 nm. Subsequently, flower extracted Au NPs showed similar

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behaviour of polydispersed nature, with the particle size varying from 10.1 to 15.6 nm. Our findings are in agreement with spherical NPs with size varying from 35-55 nm via leaf of C.

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roseus linn. g. Don has been described by Ponarulselvam et al., 2012 [23].

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3.7.2 EDAX analysis

EDAX spectrum discloses the major signal in the Ag and Au region, which reveals the

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creation of Ag and Au NPs (Fig. 6). The elemental compositions of Ag and Au NPs were 43.02 and 41.94 wt%, correspondingly. EDAX profile showed the existence of a strong

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signal for elemental Ag at ~ 3 keV, while the existence of strong absorption signal for elemental Au at ~1.5 keV. Further, EDAX spectrum also reveals the weaker oxygen peak

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signal which might be resulted from the biomolecules that are absorbed to the surface of nanoparticles, signifying the reduction of Ag/Au ions to elemental Ag or Au. Lastly, there were no major peaks detected for synthesized NP. These results further evidence the complete bio-reduction of metal ions to metal NPs as displayed in the spectrum. Mukundan et al., 2017 [14] reported the presence of a Ag and Au peak using B. tomentosa leaves are found similar to our results.

3.8 XRD analysis The diffraction pattern of plant extracted synthesized Ag and Au NPs is shown in Fig. 7. The XRD patterns at 36.70, 42.90, 63.28 and 76.40 corresponding to the lattice planes 2.45, 2.11, 1.47, 1.24 nm and 27.32, 36.72, 39.56, 42.89, 63.25, 63.25, 65.02 and 76.38 corresponding to the lattice planes 3.26, 2.44, 2.28, 2.11, 1.47, 1.43 and 1.25 nm are in consistent with the standard pattern (JCPDS 04-0783) which illustrated that the green

Journal Pre-proof synthesized Ag and Au NPs are crystalline in nature with face centered cubic crystal structure. Previously, a pattern of XRD for Ag and Au NPs was confirmed by Arumugam et al., 2017 [8] using A. indica, M. balbisiana and O. tenuiflorum extract and also by Bornali and Mohan 2013 [5] using M. balbisiana bract extract respectively.

3.9 Gas Chromatography- Mass Spectroscopy Analysis for profiling of metabolites GC MS analysis authenticated the presence of metabolites such as (3.beta.,22Z)-4H-1,3Benzodioxin-4-one,2-(1.1-methylethyl)

hexahydro

-5-methyl-4a-(2-propenyl)-,[2S-

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(2.alpha.,4a.alpha.,5.beta.,8a.beta)]- and 9,12-octadecadienoic acid(z,z)- (Fi. 8 and Table 2). Similarly 15 bioactive compounds using methanolic extract of M. acuminate colla fruit has

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been reported by Kumar et al 2017 [24]. Jordan et al 2001 [8] reported 43 volatile

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compounds in M.acuminate colla fruit.

3.10 Antibacterial activity of flower extracts, AgNps and AuNPs of Musa acuminata

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colla flowers

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The antibacterial performances of aq. extract, ethanol extract, Ag and Au NPs of M. acuminata colla flowers were tested against the ESBL gene producing bacteria such as E.

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faecalis, S.aureus, K. pneumoniae, S. typhi, E. coli, P.mirabilis, and P.aeruginosa. Subsequently, the obtained results of NPs were summarized in Table 3. All the four samples have shown good antibacterial activities against Enterococcus faecalis, Proteus mirabilis, Pseudomonas aeruginosa and Klebsiella pneumoniae. Both extracts and silver NP have moderate activity against Staphylococcus aureus. A silver nanoparticle alone has minimal antibacterial activity against Echescheria coli and Salmonella typhi. Banerjee et al ., 2014 [1] reported antibacterial activity of banana leaf against E.coli and Bacillus showing 6 and 6 mm for extract, 14 and 16 mm for Ag NPs. In microdilution (resazurin) method, AgNPs exhibited good activity for all the organisms followed by ethanol extract and aqueous extract. AuNPs showed least activity for all the organisms (Fig. 9 and Fig. 10). Saritha et al., 2019 [25] reported that leaves of T. majus ethanol extract, aqueous extract and AgNPs showed best activity against P. aeruginosa.

Journal Pre-proof 3.11 Anticancer activities of solvent extracts, Ag NPs and Au NPs from Musa acuminata colla The numerous concentrations of aqueous extract, ethanol extract, Ag and Au NPs 200,100, 50, 25, 12.5, 6.25 and 3.12 µg/ml and vehicle control (DMSO) control (without samples) were tested for anticancer activities in MCF-7 and normal Vero cell line. Normal Vero cell lines had similar effect and the 50% viability were calculated as 55.0 µg/ml, 37.5 µg/ml, 27.5 µg/ml and 35 µg/ml for Au NPs, aqueous, ethanol and Ag NPs respectively (Fig. 9 and Fig. 10). Saritha et al., 2019 [25] reported that leaves of T. majus ethanol extract, aq. extract and Ag NPs showed IC50 value at 7.5 µg/ml, 4.68 µg/ml, 2.49 µg/ml for MCF-7 cell lines and

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IC50 for normal Vero cell lines were at 6.8 µg/ml, 8.1 µg/ml and 5.3 µg/ml.

4. Conclusion

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Aqueous and ethanol extract of M. acuminata colla flowers were analysed for phyto-

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constituents by qualitative and GC-MS. Antioxidants assay were performed by DPPH scavenging and total antioxidant scavenging analysis. Aqueous extract was used to reduce

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Ag+ and Au3+ ions to Ag0 and Au0 ions for synthesizing respective NP. UV, FT-IR, XRD and SEM – EDAX were used to characterize the synthesised NP. Both extracts, biosynthesized

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AgNPs and AuNPs were tested for its activity against ESBL gene producing bacteria. Toxicity and anti cancerous activity of the NP against normal Vero and MCF-7 cell lines

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were tested. The IC50 values noted for MCF-7 and normal VERO cells treated with AuNPs 30.0 µg/ml and 55.0µg/ml, aqueous 10.62µg/ml and 35µg/ml, ethanol 8.75 µg/ml and 27.5 µg/ml AgNps 4.04 µg/ml and 37.5 µg/ml respectively. From the perusal of the above results, the solvent extracts and NP are found to be promising to treat ESBL gene producing bacteria as well as anticancer activities by the pharmaceutical industries.

Acknowledgement The authors SV and PA thank Loyola College, Madras University Chennai and Royal Bio Research Centre, Chennai for providing lab facilities to carry out this research work. The authors MVA, NAA-D, GAE and AKMG would like to extend their sincere appreciation to the Deanship of Scientific Research at King Saud University for its funding of this research through the Research Group Project No. RG-1435-071.

References

Journal Pre-proof [1] P. Banerjee, M. Satapathy, A. Mukhopahayay, P. Das, Leaf extract mediated green synthesis of silver nanoparticles from widely available Indian plants: synthesis, characterization,

antimicrobial

property

and

toxicity

analysis,

Bioresources

and

Bioprocessing, 1 (2014) 3. [2] M. Kavitha, A.C. Ponni, World Journal of Science and Research, World, 2 (2017) 18-27. [3] H.M. Ibrahim, Green synthesis and characterization of silver nanoparticles using banana peel extract and their antimicrobial activity against representative microorganisms, Journal of Radiation Research and Applied Sciences, 8 (2015) 265-275. [4] S. Saranya, A. Eswari, E. Gayathri, S. Eswari, K. Vijayarani, Green synthesis of metallic

of

nanoparticles using aqueous plant extract and their antibacterial activity, Int. J. Curr.

ro

Microbiol. App. Sci, 6 (2017) 1834-1845.

[5] B. Bornali, M. Kalita, Green synthesis of gold nanoparticles using Musa balbisiana bract

-p

extract, International Journal of Pharma and Bio Sciences, 4 (2013). [6] C. Gomathi, Synthesis and characterization of zinc oxide nanoparticles using aqueous

re

extract of Banana Peel (Musa acuminata L.) R. Ananthalakshmi*, SR Xavier Rajarathinam2, A. Lavanya1, A. Mohammed Sadiq3 and A.

lP

[7] Al-Dhabi NA, Ghilan AKM, Arasu MV, Duraipandiyan V, Ponmurugan K. Environmental friendly synthesis of silver nanomaterials from the promising Streptomyces

na

parvus strain Al-Dhabi-91 recovered from the Saudi Arabian marine regions for antimicrobial

189:176-184.

Jo ur

and antioxidant properties. Journal of Photochemistry & Photobiology, B: Biology: 2018a:

[8] N. Arumugam, B. Thulasinathan, R. Pasubathi, K. Thangavel, J.B. Muthuramalingam, A. Arunachalam, Biogenesis of silver nanoparticles using selected plant leaf extract; characterization and comparative analysis of their antimicrobial activity, Nanomedicine Journal, 4 (2017) 208-217.

[9] J.S. Moodley, S.B.N. Krishna, K. Pillay, P. Govender, Green synthesis of silver nanoparticles from Moringa oleifera leaf extracts and its antimicrobial potential, Advances in Natural Sciences: Nanoscience and Nanotechnology, 9 (2018) 015011. [10] S. Dahham, Y. Tabana, D. Sandai, M. Ahmed, A. Majid, In vitro anticancer and antiangiogenic activity of essential oils extracts from agarwood Aquilaria crassna, Med. Aromat. Plants, 5 (2016) 256-268. [11] S.S. Dahham, T. Mohamad, Y.M. Tabana, A. Majid, Antioxidant activities and anticancer screening of extracts from banana fruit (Musa sapientum), Academic J Cancer Res, 8 (2015) 28-34.

Journal Pre-proof [12] L. Leaves, Antioxidant activity by DPPH radical scavenging method of ageratum conyzoides, American Journal of Ethnomedicine, 1 (2014) 244-249. [13] A.B. Aliyu, M.A. Ibrahim, A.M. Musa, A.O. Musa, J.J. Kiplimo, A.O. Oyewale, Free radical scavenging and total antioxidant capacity of root extracts of Anchomanes difformis Engl.(Araceae), Acta Pol Pharm, 70 (2013) 115-121. [14] Al-Dhabi NA, Esmail GA, Duraipandiyan V, Valan Arasu M, Salem-Bekhit MM. Isolation, identification and screening of antimicrobial thermophilic Streptomyces sp. AlDhabi-1 isolated from Tharban hot spring, Saudi Arabia. Extremophiles 2016;20:79-90. [15] Balachandran C, Duraipandiyan V, Emi N, Ignacimuthu S. Antimicrobial and cytotoxic

of

properties of Streptomyces sp. (ERINLG-51) isolated from SouthernWestern Ghats. South

ro

Indian J Biol Sci 2015;1:7–14.

[16] Al-Dhabi NA, Esmail GA, Duraipandiyan V, Valan Arasu M. Chemical profiling of

-p

Streptomyces sp. Al-Dhabi-2 recovered from an extreme environment in Saudi Arabia as a novel drug source for medical and industrial applications. Saudi J Biol Sci 2019a;26:758-66.

re

[17] V. Sumathy, S.J. Lachumy, Z. Zakaria, S. Sasidharan, In vitro bioactivity and phytochemical screening of Musa acuminata flower, Pharmacologyonline, 2 (2011) 118-127.

lP

[18] O. Obiageli, Izundu A. I.,Okoye Nkechi Helen,I.Adachukwu Pauline. Phytochemical Compositions of Fruits of Three Musa Species at Three Stages of Development. IOSR

na

Journal of Pharmacy and Biological Sciences 2016, 48-59. [19] Al-Dhabi NA, Esmail GA, Ghilan AKM, Arasu MV, Duraipandiyan V, Ponmurugan K.

Jo ur

Chemical constituents of Streptomyces sp. strain Al-Dhabi-97 isolated from the marine region of Saudi Arabia with antibacterial and anticancer properties. Journal of Infection and Public Health. 2019: https://doi.org/10.1016/j.jiph.2019.09.004 [20] M.Z. Imam, S. Akter, M.E.H. Mazumder, M.S. Rana, Antioxidant activities of different parts of Musa sapientum L. ssp. sylvestris fruit, Journal of Applied Pharmaceutical Science, 1 (2011) 68. [21] D. Mukundan, R. Mohankumar, R. Vasanthakumari, Green synthesis of silver nanoparticles using leaves extract of Bauhinia tomentosa linn and its invitro anticancer potential, Materials Today: Proceedings, 2 (2015) 4309-4316. [22] Ilavenil, S., Srigopalram S., Park, HS., Choi KC. 2015. Growth and metabolite profile of Pediococcus pentosaceus and Lactobacillus plantarum in different juice. South Indian Journal of Biological Science, 1, 1-6. [23] S. Ponarulselvam, C. Panneerselvam, K. Murugan, N. Aarthi, K. Kalimuthu, S. Thangamani, Synthesis of silver nanoparticles using leaves of Catharanthus roseus Linn. G.

Journal Pre-proof Don and their antiplasmodial activities, Asian Pacific journal of tropical biomedicine, 2 (2012) 574-580. [24] Kumar D, Punithavel N, Jayachandran M. Green chemical approach towards the synthesis of CeO2 doped with seashell and its bacterial applications intermediated with fruit extracts. Journal of Photochemistry & Photobiology, B: Biology: 2017: 172: 50-60. [25] Valsalam S, Agastian P, Arasu MV, Al-Dhabi NA, Ghilan AKM, Kaviyarasu K, Ravindran B, Chang SW, Arokiyaraj S. Rapid biosynthesis and characterization of silver nanoparticles from the leaf extract of Tropaeolum majus L. and its enhanced in-vitro antibacterial, antifungal, antioxidant and anticancer properties. Journal of Photochemistry &

Jo ur

na

lP

re

-p

ro

of

Photobiology, B: Biology: 2019: 191:65-74.

Journal Pre-proof Table 1: Phytochemical analysis of solvent extract of Musa acuminate colla

Contents

Aqueous extract

Ethanol extract

Tannins

+

-

2.

Saponins

-

-

3.

Flavonoids

+

-

4.

Alkaloids

+

+

5.

Proteins

+

+

6.

Steroid

+

+

7.

Quinones

+

-

8.

Terpenoids

-

-

9.

Cardio glycosides

+

+

10.

Phenol

-

-

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1.

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S.No

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+: the components are present; -: the components are absent

Journal Pre-proof Table 2: The major phytochemical compounds identified from the ethanol extract of Musa acuminate colla flower by GC–MS. RI

List of compounds

3.48 6.98 17.43 1.12 1.11 2.44 2.48 1.42 1.68 1.21 3.67 4.16 7.73 21.31 21.16

35.880

2.63

Phenol,5-(1,5-dimethyl-4-hexenyl) -2-methyln-hexadecanoic acid 9,12-octadecadienoic acid(z,z)Octacosane Tetracosane Tetracosane Tetracosane Tritriacontane Octacosane Octacosane 3,5-Dihydro-3-((1-methylethyl) imino) –N, Stigmasterol .beta.-Sitosterol Curan-17-oic acid, 2,16-didehydro- 20-hydroxy-19-oxo9-Isopropyl-6-methyl-6,7-dihydro-9 H-5-oxa-9-azabenzocyclohepten-8-on Cycloheptane 9,19-cyclolanostan-3-ol,

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Retention tine 16.742 18.960 20.614 23.735 24.753 26.005 27.391 28.782 30.145 31.484 32.905 33.260 34.034 34.709 34.907

Journal Pre-proof Table 3: Antibacterial activity of Musa acuminata colla flower of ethanol extract against ESBL gene producing bacteria

S.No Microorganisms/Sample

Zone of inhibition(mm) 1000 µg

500 µg

250 µg

125 µg

DMSO

Streptomycin (Std 10µg)

Aqueous Extract Enterococcus faecalis

10

9

9

8

-

14

2

Staphylococcus aureus

11

9

7

-

-

22

3

Klebsiella pnumoniae

12

10

6

-

-

16

4

Salmonella typhi

-

-

-

-

-

15

5

Echerichia coli

7

-

-

-

-

16

6

Proteus mirabilis

12

10

9

6

-

19

7

Pseudomonas aeruginosa

9

8

6

-

-

13

10

-

-

14

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Enterococcus faecalis

13

6

2

Staphylococcus aureus

8

6

7

6

-

22

3

Klebsiella pnumoniae

7

-

-

-

-

16

4

Salmonella typhi

6

6

-

-

-

15

5

Echerichia coli

-

-

-

-

-

16

6

Proteus mirabilis

13

12

10

10

-

19

7

Pseudomonas aeruginosa

8

7

7

6

-

14

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Ethanol Extract

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1

AgNPs 1

Enterococcus faecalis

13

10

9

7

-

14

2

Staphylococcus aureus

9

-

-

-

-

22

3

Klebsiella pnumoniae

10

9

-

-

-

16

4

Salmonella typhi

9

7

6

-

-

15

5

Echerichia coli

12

10

8

-

-

16

6

Proteus mirabilis

6

6

6

-

-

19

Journal Pre-proof 7

Pseudomonas aeruginosa

12

10

8

7

-

13

AuNPs Enterococcus faecalis

11

8

8

6

-

14

2

Staphylococcus aureus

-

-

-

-

-

22

3

Klebsiella pnumoniae

10

-

-

-

-

16

4

Salmonella typhi

9

-

-

-

-

15

5

Echerichia coli

7

-

-

-

-

16

6

Proteus mirabilis

8

7

7

6

-

19

7

Pseudomonas aeruginosa

9

8

8

-

13

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1

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Fig.1: Antioxidant activity of various extract of Musa acuminata colla DPPH assay and Total antioxidant assay

Journal Pre-proof A

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Fig 2. Synthesis of silver and gold nanoparticles from the flower extract of Musa acuminata colla A=AgNPs, 1=Silver nitrate, 2=before synthesis, 3= after synthesis B=AuNPs, 1=Gold chloride, 2=before synthesis, 3= after synthesis B

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Fig 3. UV–visible spectrum of the (A) silver and (B) gold nanoparticles synthesized from the flower extract of Musa acuminata colla

Fig 4. FTIR spectrum of the synthesized from the flower extract of A= Musa acuminata colla flower extract, B=AuNPs, C= AgNPs

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A1

B1

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Fig 5. SEM image of the synthesized (A,A1) silver (B,B1) gold nanoparticles from the flower extract of Musa acuminata colla

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Fig 6. SEM -EDAX image of the synthesized (A) silver and (B) gold nanoparticles from the flower extract of Musa acuminata colla

B

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30000

B

A 10000

Intensity (cps)

Intensity (cps)

20000

5000

10000

0

0 20

40

60

20

80

40

60

80

2-theta (deg)

2-theta (deg)

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Fig 7. XRD pattern of the synthesized (A) silver and (B) gold nanoparticles from the flower extract of Musa acuminata colla

Fig.8: GC-MS analysis of the ethanolic extract of Musa acuminate colla

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Fig 9. IC50 values of Musa acuminata colla flower extracts against MCF-7cell lines. a. Aqueous extract, b. Ethanol extract, c. Silver nanoparticles, d. Gold nanoparticles, e. Vehicle control (DMSO) and f. Control against MCF-7cell lines.

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b \ b

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Fig 10. IC50 values of Musa acuminata colla flower extracts against VERO cell lines. a. Aqueous extract, b. Ethanol extract, c. Silver nanoparticles, d. Gold nanoparticles, e. Vehicle control (DMSO) and f. Control against VERO cell lines.

Journal Pre-proof Conflict of Interest Statement I would like to inform you that the authors in manuscript entitled “Biosynthesis of Silver and Gold nanoparticles using Musa acuminata colla flower and its pharmaceutical activity against bacteria and anticancer efficacy” do not have any conflict of interest in submitting this paper to Journal of Photochemistry and Photobiology B

Regards

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Dr. Mariadhas Valan Arasu

Journal Pre-proof Highlights  Ag and Au nanoparticles were green synthesized from the flower extract of M.acuminata colla  Nanoparticles showed maximum absorption at 474 nm and 540 nm  TEM and SEM analysis confirmed the nanoparticels were lattice plane  Nanoparticles exhibited activity against Pseudomonas aeroginosa

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 Nanoparticles documented both antioxidant and anticancer properties