Biosynthesis, Characterization of Gold Nanoparticles

International Journal of Nanobiotechnology ISSN: 2456-0111 (online) Vol. 2: Issue 2

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Biosynthesis, Characterization of Gold Nanoparticles Using M. indica Leaf Extract and Their Anticancer Activity Nabeel Ahmad1, Syed Mohd. Danish Rizvi2, Nitin Sahai3, Rajiv Dutta4* 1

Department of Biotechnology, School of Engineering and Technology, IFTM University, Lodhipur Rajput Moradabad, UP, India 2 Department of Biosciences, Integral University, Lucknow, UP, India 3 Department of Biomedical Engineering, North Eastern Hill University, Shillong, Meghalaya, India 4 Department of Biotechnology, Sharda University, Greater Noida, UP, India

ABSTRACT The present study involves the novel approach for the synthesis of nanomaterial holds infinite possibilities as nanotechnology has a major impact on medicine and its application. Leaf extract of Mangifera indica is a very good bioreductant for gold chloride (HAuCl4·4H2O) which leads to the synthesis of gold nanoparticles. Leaf extract of Mangifera indica was mixed with gold chloride (HAuCl4·4H2O) and UV–Vis spectroscopy was used to detect the synthesis of nanoparticles. TEM and FESEM were used for size, shape and structural morphology of synthesized nanoparticles. Synthesized gold nanoparticles were in range of less than ~75 nm and hexagonal in shape. The gold nanoparticles also showed potent cytotoxic effect against A549 lung cancer cell lines with an IC 50 value of 59 μg/ml by the MTT assay. Keywords: biosynthesis, characterization, cytotoxicity gold chloride, nanotechnology *Corresponding Author E-mail: [email protected]

INTRODUCTION Current development in nanotechnology lead to the fabrication of engineered metallic nanoparticles those are capable and applicable towards human development. Metallic nanoparticles have been shown to reveal unusual properties that they didn’t exhibit in their bulk in form. Because of these singular properties, nanoparticles applications are being researched extensively and these materials are edging the conventional materials in various purposes across wide range of disciplines. Metallic nanoparticles such as gold, silver, copper and zinc are being used as catalysts, chemical and biological sensors in several biomedical applications. In medicine, they are likely to be applied as antimicrobial, antifungal and as drug delivery agents. Metallic nanoparticles are

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the most promising because of their high surface to volume ratio.[1–3] Several methods like physical and chemicals are available for the synthesis of metallic nanoparticles but they are very hazardous, toxic, costly vacuum system, environment containment and time taking process.[4] Thus, the green synthesis bioprocess is much needed and hence has been finding widespread interest. The biosynthesis of metallic nanoparticles from corresponding metals carries out several routes like plant extracts, enzymes and using micro- organisms. The potential of plants to synthesis nanoparticles have featured excitingly very large toward the increased of natural nano-factories.[5,6] This remarkable development has brought an insight among researchers for

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Biosynthesis, Characterization of Gold Nanoparticles

advancement of molecules which are precise and efficient by using refined and potentially remarkable strategies. The main aim of this study was to synthesize AuNPs using aqueous leaves extract of Mangifera indica as bioreducing agent to reduce Au+ to Au0, which were later analyzed and characterized by using ultraviolet–visible (UV–Vis) spectroscopy, transmission electron microscopy (TEM) and field emission scanning electron microscopy (FESEM). After characterization, the in vitro cytotoxic activities of the green synthesized AuNPs were evaluated. MATERIAL AND METHODS Reagents Chloroauric acid (HAuCl4·4H2O) was obtained from CDH Pvt Ltd and used as received. Collection, Plant Extract Preparation and Biosynthesis of Gold Nanoparticles Fresh and healthy leaves of Mangifera indica were harvested from garden of Department of Biotechnology, IFTM University, Moradabad, Uttar Pradesh, India. Leaves of Mangifera indica were washed with distilled water, cut into fine pieces. Ten grams of fine cut leaves were added in 100 mL of distilled water and boiled for 20 minutes in agar heating mantle at 70°C. This mixture was cooled to room temperature and then filtered using Whatman filter paper. AuNPs were synthesized according to the procedure described in previous study.[7] Typically, 5 mL of Mangifera indica plant extract was added to 45 mL of 1 mM aqueous HAuCl4 solution for the reduction of Au3+ to Au0. As time passed, change of color of solution changed from pale yellow to violet red, which confirms that, the synthesis of AuNPs.


Ahmad et al.

Characterization of AuNPs The synthesized AuNPs were characterized by UV–Vis spectroscopy, TEM, and FESEM. Cell Culture and MTT Assay A549 cell line was procured from National Centre for Cell Science, Pune, India. The cell lines were grown as a monolayer in DMEM medium. To determine the cytotoxic effect of nanoparticles, cell viability study was done with the conventional MTT-reduction assay with slight modifications (Chencharick and Mossman, 1983).[8] RESULT AND DISCUSSION Gold Nanoparticles reveals violet red color in aqueous solution due to excitation of Surface Plasmon Resonance, which is a unique trait of AuNPs. The appearance of the violet red color was only due to the reduction of Au+, which confirms the formation of Au nanoparticles as shown in Figure 1.

Fig. 1. Synthesis of gold nanoparticles. The UV–Vis spectrum of AuNPs synthesized by Mangifera indica is shown in Figure 2. The distinct peak observed at ~550 nm corresponds to surface plasmon resonance of AuNPs.[9,10]

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Fig. 2. UV–Vis analysis of gold nanoparticles. TEM technique was used to visualize the size and shape of synthesized AuNP. It is evident from Figure 3, that most of the synthesized particles were uniform polydispersed, irregular and hexagonal in shape. The average size of the individual nanoparticles was estimated ~34 nm,

Fig. 3. TEM image Toxicological analysis is one of the cornerstones for the application of nanoparticles in medicinal applications.[11] The study found that gold nanoparticles affect cancerous cell lines in a dose

whereas polydispersed particles were in the range of 50–75 nm. The surface morphology of AuNPs was investigated using FESEM. The micrographs (Figure 4) showed that synthesized AuNPs have rough surfaces.

Fig. 4. FESEM image. dependent manner. The cell viability was found to be decreasing almost with concentration of 59 μg AuNPs in the case of A549 as shown in Figure 5A and B.

Fig. 5. (A) Phase contrast microscopic images of AuNPs induced gross cytomorphological changes and growth inhibition on A549 cells. (B) Effect of AuNPs on cytotoxicity in A549 cells. Data represent mean ± S.E. of three experiments.


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CONCLUSION Nanotechnology is facilitating technology that deals with nanometer sized items. The exercise of nanomaterials in biotechnology unites the fields of biology and material science. This method was very facile for the synthesis of gold nanoparticles. Synthesized gold nanoparticles were characterized by using UV–Vis spectroscopy, TEM and FESEM. The average size of the individual nanoparticles was estimated ~34 nm. Cytotoxic activity of gold nanoparticles was characterized by the means of a cell viability assay on A549 cancerous cell lines. ACKNOWLEDGMENT We would like to extend our gratitude to the SAIF Chandigarh, India for characterization of sample. REFERENCES [1] Ahmad N., Shree K., Srivastava M., et al. Novel rapid biological approach for synthesis of silver nanoparticles and its characterization, Int J Pharmacol Pharm Sci. 2014; 1: 28–31p. [2] Song J.Y., Jang H.K., Kim B.S. Biological synthesis of gold nanoparticles using Magnolia kobus and Diopyros kaki leaf extracts, Process Biochem. 2009; 44(10): 1133–8p. [3] Kumar V., Yadav S.K. Plant‐mediated synthesis of silver and gold nanoparticles and their applications, J Chem Technol Biotechnol. 20091; 84(2): 151–7p.


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Goodman C.M., McCusker C.D., Yilmaz T., et al. Toxicity of gold nanoparticles functionalized with cationic and anionic side chains, Bioconjug Chem. 2004; 15(4): 897– 900p. [5] Gan P.P., Li S.F. Potential of plant as a biological factory to synthesize gold and silver nanoparticles and their applications, Rev Environ Sci Bio/Technol. 2012; 11(2): 169–206p. [6] Ahmad N., Bhatnagar S., Ali S.S., et al. Phytofabrication of bioinduced silver nanoparticles for biomedical applications, Int J Nanomed. 2015; 10: 7019p. [7] Mubarak Ali D., Thajuddin N., Jeganathan K., et al. Plant extract mediated synthesis of silver and gold nanoparticles and its antibacterial activity against clinically isolated pathogens, Colloids and Surfaces B: Biointerfaces. 2011; 85(2): 360–5p. [8] Chencharick J.D., Mossman K.L. Nutritional consequences of the radiotherapy of head and neck cancer, Cancer. 1983; 51(5): 811–5p. [9] Philip D., Unni C. Extracellular biosynthesis of gold and silver nanoparticles using Krishna tulsi (Ocimum sanctum) leaf, Physica E. 2011; 43(7): 1318–22p. [10] Narayanan K.B., Sakthivel N. Coriander leaf mediated biosynthesis of gold nanoparticles, Mater Lett. 2008; 62(30): 4588–90p. [11] Pan Y., Neuss S., Leifert A., et al. Size‐dependent cytotoxicity of gold nanoparticles, Small. 2007; 3(11): 1941–9p.

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