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[133] Elumalai, E. K., Prasad, T. N. V. K. V., Hemachandran, J., Viviyan, Therasa, S., Thirumalai, T., and E. David, “Extracellular synthesis of silver nanoparticles using leaves of Euphorbia Hirta and their antibacterial activities”, Journal of Pharmaceutical Sciences and Research, 2010, 2(9), p. 549.

[134] Krishnaraj, C., Jagan, E. G., Rajasekar, S., Selvakumar, P., Kalaichelvan, P. T., and N. Mohan, “Synthesis of silver nanoparticles using Acalypha indica leaf extracts and Its antibacterial activ ity against water borne pathogens”, Colloids and Surfaces B: Biointerfaces, 2010, 76, pp. 50–56.

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[142] Avinash Kumar Reddy, G., Jyothi M. Joy, Trilok Mitra, Shaik, Shabnam, and T. Shilpa, “Nano Silver – A Review”, International Journal of Advanced Pharmaceutics, 2012, 2(1), pp. 9–15.

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[148] Url: www.vtek.chalmers.se/~v92tilma/tea/misc/chem.html.[149] Sun, R. W. Y., Chen, R., Chung, N. P.Y., Ho, C.M., Lin, C.L.

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[153] Paulina Segovia, Víctor Coello, Jesús Arriaga, Sergio Belmares, Aracelia Alcorta, Francisco Hernández, Ricardo Obregón, Ernesto Torres, and Francisco Paraguay, “In TechGreen synthesis and char acterizations of silver and gold nanoparticles”, Univer sidad Autónoma de Nuevo León, San Nicolás de los Garza, N. L., México.

[154] Kerker, M., “The optics of colloidal silver: Something old and something new”, J Colloid Interface Sci, 1985, 105, pp. 297–314.

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[156] Lok, C., “Proteomic analysis of the mode of antibacterial action of silver nanoparticles”, J Proteome Res, 2006, 5, pp. 916–924.

[157] Jiang, X. C., Chen, W. M., Chen, C. Y., Xiong, S. X., and A. B. Yu, “Role of temperature in the growth of silver nanoparticles through a synergetic reduction approach”, Nanoscale Res Lett, 2011, 6(1), p. 32.

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[159] Raffi, M., Hussain, F., Bhatti, T. M., Akhter, J. I., Hameed, A., and M. M. Hasan, “Antibacterial characterization of silver nanoparticles against E. Coli ATCC5224”, J Mater Sci Technol, 2008, 24(2), pp. 97–192.

[160] Jiang, X. C., Chen, W. M., Chen, C. Y., Xiong, S. X., and A. B. Yu, “Role of temperature in the growth of silver nanoparticles through a synergetic reduction approach”, Nanoscale Res Lett, 2011, 6, 32.

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List of figures

Figure 3-1 (a, b, c)

Pictures of black tea leaves, garlic and onion extract respectively. 20

Figure 3-2(a, b)

Schema of experimental setup and picture of experimental setup for electrolytic deposition of silver nanoparticles using different values of cur-rent in mA respectively. 21

Figure 3-3(a, b)

Sketch of the experimental setup and picture showing setup in lab respectively. 23

Figure 3-4 Sketch of experimental set up for method III. 25Figure 3-5 Picture of the experimental set up for method

III in the institute lab. 25Figure 3-6(a, b)

Sketch of the experimental setup and picture of the experimental setup in the institute lab for method IV. 26

Figure 3-7 (a, b)

Sketch of the experimental setup and picture of the actual set up in the institute lab respectively for method IV. 27

Figure 4-1 (a–d)

Changes in the colour of the solution during elec-trolysis. (a) Colour of the solution immediately after the circuit is switched on and tea extract is added. (b) Colour of the solution after 5 min. (c) Colour of the solution after 10 min. (d) The particles collected from the cathode and settled down at the bottom of the surface after 48 hrs. 30

Figure 4-2 Samples corresponding to 50 mA, 150 mA, 200 mA, 250 mA, 350 mA, 500 mA current values (from left to right). 31

Figure 4-3 (a–g)

XRD of as-synthesized silver nanoparticles cor-responding to (a) 500 mA, (b) 350 mA, (c) 250 mA, (d) 200 mA, (e) 150 mA, (f ) 50 mA and (g) JCPDS file No. 04-0783. 32

Figure 4-4 (a–d)

TEM picture showing the particles of size 32–97 nm for current 59 mA. 33

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TEM photograph showing the particles of size 30 nm–80 nm for current 150 mA. 34

Figure 4-6 (a–e)

TEM photograph showing the particles of size 12 nm–70 nm for current 200 mA. 34


TEM photograph showing the particles of size 15–35 nm for current 250 mA. 35


TEM photograph showing the particles of size 5–35 nm for current 350 mA. 35


TEM image of silver nanoparticles correspond-ing to 500 mA. 36

Figure 4-10 UV-vis graphs of as-synthesized silver nanopar-ticles for set 1 of method 1. 37

Figure 4-11 Picture showing the final colour of the solu-tions obtained for 1.25%, 2.5% and 6.25% (v/v) concentration of black tea leaves extract. 38


XRD graphs for 1.25%, 2.5% and 6.25% (v/v) concentration of black tea leaves extract respectively. 39

Figure 4-13 (a, b)

TEM pictures of silver nanoparticles of size 12 nm–70 nm for 1.25% (v/v) concentration of black tea leaves extract. 40


TEM pictures of silver nanoparticles of size 2 nm–40 nm for sample with 2.5% (v/v) con-centration of black tea leaves extract. 41


TEM pictures of silver nanoparticles of size 2 nm–35 nm for sample with 6.25% (v/v) con-centration of black tea leaves extract. 41

Figure 4-16 UV-visible graph of as-synthesized silver nanoparticles corresponding to 1.25%, 2.5% and 6.25% (v/v) concentration of black tea leaves extract. 42

Figure 4-17 (a–h)

Pictures of inhibition zones obtained in the agar plates with organisms, antibiotics and as-synthesized silver nanoparticles. 46

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List of Figures 153

Figure 4-18 Dose response curve for compounds against MCF-7 cell line. 47

Figure 4-19 Dose response curve for compounds against He-La cell line. 48

Figure 4-20 Colour of colloidal silver for different strengths of precursor. 50


XRD graphs corresponding to 0.005N, 0.01N and 0.02N strength of silver nitrate. 51

Figure 4-22 (a, b)

TEM pictures of as-synthesized silver for 0.005N silver nitrate solution. 52


TEM pictures of as-synthesized silver nanopar-ticles for 0.01N strength. 52

Figure 4-24 (a, b)

TEM and AFM pictures of the silver nanopar-ticles with size >100 nm. 53

Figure 4-25 UV-visible graphs of silver nanoparticles corre-sponding to different strengths of precursor. 53

Figure 4-26 Picture of coloured solution obtained for 1.25%, 2.5% and 6.25% (v/v) concentration respectively of capping agent for 0.005N of precursor. 54


XRD graphs of as-synthesized silver nanoparticles corresponding to 1.25%, 2.5% and 6.25% (v/v) concentration of the capping agent respectively. 55

Figure 4-28 TEM picture of as-synthesized silver nanopar-ticles of size 5–60 nm for 1.25% (v/v) concen-tration of capping agent with 0.005N strength of silver nitrate. 56

Figure 4-29 (a, b)

TEM picture of as-synthesized silver nanopar-ticles of size 2–30 nm for 2.5% (v/v) concentra-tion of capping agent with 0.005N strength of silver nitrate. 56

Figure 4-30 (a, b)

TEM picture of as-synthesized silver nanopar-ticles of size 2–25 nm for 6.25% (v/v) concen-tration of capping agent with 0.005N strength of silver nitrate. 57



Figure 4-31 UV-visible graph for 2.5% and 6.25% (v/v) concentration for 0.005N strength of silver nitrate. 57

Figure 4-32 (a, b)

FTIR graph as-synthesized silver nanoparticles capped by black tea leaves extract respectively. 58

Figure 4-33 Dose response curve for as-synthesized silver nanoparticles against MCF-7 cancer cell line. 59

Figure 4-34 Dose response curve of as-synthesized silver nanoparticles against He-La cancer cell line. 60

Figure 4-35 Pictures of the coloured solution of the samples for black tea leaves, garlic and onion extract. 61

Figure 4-36(a, b, c)

XRD graphs for three different capping agents.61


TEM and AFM images of as-synthesized silver nanoparticles of size 12–70 nm for tea as cap-ping agent. 62


TEM pictures of silver nanoparticles of size 5–25 nm in case of onion extract as capping agent. 63

Figure 4-39 (a–e)

TEM images of silver nanostructures for 0.0125 of garlic extract. 63

Figure 4-40 UV-visible graphs of as-synthesized silver nanoparticles for different capping agents. 64

Figure 4-41 Colour of the solution obtained for 1.25%, 2.5%, 6.25% (v/v) concentration of onion extract for 0.005N silver nitrate. 65


XRD graphs of as-synthesized silver nanoparti-cles for 1.25%, 2.5%, 6.25% (v/v) concentration of onion extract. 65


TEM images of silver nanoparticles of size 5–25 nm for 1.25% (v/v) concentration of onion extract. 66


List of Figures 155


TEM images of silver nanoparticles synthesized of size 2–20 nm using 2.5% (v/v) concentration of onion extract as capping agent. 66


TEM and AFM images of silver nanoparticles of size 2–10 nm for 6.25% (v/v) concentration of onion extract as capping agent respectively. 67

Figure 4-46 UV-graphs of silver nanoparticles for different concentration of onion extract. 68

Figure 4-47 (a–f )

Inhibition zones for silver nanoparticles in com-bination with antibiotics. 69

Figure 4-48 Colour of the solutions of the samples corre-sponding to different temperatures for method II. 74


XRD graphs of as-synthesized silver nanopar-ticles at 30°C, 50°C, 70°C. 75


TEM pictures of as-synthesized silver nanopar-ticles of size 2–35 nm from method II at 30°C. 76

Figure 4-51 (a–e)

TEM pictures of silver nanoparticles of size 30–100 nm from tollens method at 50°C. 76


TEM pictures of silver nanoparticles from tol-lens method at 70°C. 77

Figure 4-53 UV-visible graphs of silver nanoparticles for precursor solution at 30°C, 50°C, 70°C temperatures. 78

Figure 4-54 FTIR spectrum of pure black tea leaves. 79Figure 4-55 FTIR spectrum of the pure black tea leaves and

the as-synthesized silver nanoparticles at 30°C. 80Figure 4-56 FTIR spectrum of as-synthesized silver

nanoparticles with black tea leaves extract at 50°C. 80

Figure 4-57 FTIR spectrum of as-synthesized silver nanoparticles with black tea leaves extract at 70°C. 81



Figure 4-58 Colour of samples corresponding to 1.25%, and 2.5%, 6.25% (v/v) concentrations of capping agent. 81


XRD graphs of as-synthesized silver nanopar-ticles for different concentrations of capping agent using method II. 82

Figure 4-60 (a, b)

TEM pictures of as-synthesized nanoparticles for 1.25% concentration of the capping agent. 83

Figure 4-61 (a–e)

TEM picture of as-synthesized silver nanopar-ticles of size 18–19 nm, using second method for 2.5% (v/v) concentration of capping agent. 83

Figure 4-62 (a, b)

TEM pictures of as-synthesized particles of size 2–20 nm using 6.25% (v/v) of capping agent. 84

Figure 4-63 UV-visible graphs of silver nanoparticles for method II corresponding to 1.25%, 2.5%, 6.25% (v/v) concentration of capping agent. 84

Figure 4-64 Colour of the samples obtained for different capping agents. 85


XRD patterns of as-synthesized silver nanopar-ticles using black tea leaves, garlic and onion extract. 86

Figure 4-66 (a, b)

TEM pictures of silver nanoparticles of size 2 nm–45 nm using 1.25% (v/v) black tea leaves extract. 86


TEM picture of silver nanoparticles of size 2–30 nm as-synthesized 1.25°% (v/v) onion extract in method II. 87


TEM pictures of as-synthesized nanoparticles of the size 2–50 nm using method II for garlic extract. 87

Figure 4-69 UV-visible graphs of silver nanoparticles for dif-ferent capping agent in method II. 88

Figure 4-70 XRD pattern of silver nanoparticles obtained from photographic films. 90


List of Figures 157


HRSEM of silver nanoparticle obtained from photographic films. 90

Figure 4-72 Colour of the colloidal silver solution. 91Figure 4-73 XRD graph of silver nanoparticles obtained

from X-Ray films in broad daylight without capping agents. 92

Figure 4-74(a–d)

TEM pictures of silver nanoparticles obtained from X-Ray films without using capping agents. 92

Figure 4-75 UV-visible graph for the sample obtained from X-Ray films. 93

Figure 4-76 XRD pictures of sample containing silver nanoparticles by using X-Ray films. 94

Figure 4-77(a–f )

TEM pictures of silver nanoparticles obtained from X-Ray films. 95

Figure 4-78 Colour of the solution containing silver nanoparticles obtained using X-Ray films and 1.25%, 2.5%, 6.25% (v/v) concentration of black tea leaves extract. 95


XRD patterns of silver nanoparticles obtained from X-Ray films for 1.25%, 2.5%, 6.25% (v/v) concentration of black tea leaves extract respectively. 96


TEM pictures of silver nanoparticles of size 30 nm–100 nm obtained from X-Ray films through electrolysis for 1.25% (v/v) concentra-tion of black tea leaves extract. 96


TEM images of silver nanoparticles obtained from X-Ray films using electrolysis and 2.5% (v/v) concentration of black tea leaves extract. 97

Figure 4-82 (a–d)

TEM pictures of silver nanoparticles obtained from X-Ray films through electrolysis and using 2.5% (v/v) concentration of black tea leaves extract and silver nanoparticles depositing on the bacteria. 98




TEM pictures of silver nanoparticles obtained from X-Ray films using electrolysis and 6.25% (v/v) black tea leaves extract. 99

Figure 4-84 UV-visible graphs of silver nanoparticles for three different concentration of the capping agent. 99

Figure 4-85 Colour of the samples obtained from X-Ray films for different capping agents. 100


XRD graphs of silver nanoparticles obtained from X-Ray films using NaOH as stripping agent and different capping agents: onion, garlic and tea extract. 101


TEM pictures of silver nanoparticles of size 30–100 nm obtained from X-Ray films using electrolysis and 2.5% (v/v) tea leaves extract. 101


TEM pictures of silver nanoparticles of size 100–150 nm obtained from X-Ray films capped by 1.25% (v/v) onion extract. 102


TEM images of silver nanoparticles obtained from X-Ray films through electrolysis and using 1.25% (v/v) concentration of garlic extract. 102

Figure 4-90 UV-visible graphs of silver nanoparticles obtained from X-Ray films using 1.25% (v/v) concentration of black tea leaves, onion and garlic extract. 103

Figure 4-91 Colour of colloidal silver obtained from X-Ray films using NaOH and different concentration of capping agents. 104


XRD patterns of silver nanoparticles obtained by stripping X-Ray films using NaOH for 1.25%, 2.5% and 6.25% (v/v) concentration of black tea leaves extract. 105


List of Figures 159


TEM images of silver nanoparticles of size 10–150 nm obtained from X-Ray films using NaOH and 1.25% (v/v) concentration of black tea leaves extract. 106


TEM images of silver nanoparticles of size 5–150 nm obtained from X-Ray films using NaOH and 2.5% (v/v) concentration of black tea leaves extract. 106


TEM pictures of silver nanoparticles of size 60–300 nm obtained from X-Ray films using NaOH and 6.25% (v/v) concentration of black tea leaves extract. 107

Figure 4-96 UV-visible graphs of silver nanoparticles of size 60–300 nm obtained from X-Ray films using NaOH. 108

Figure 4-97 Colour of the samples of silver nanoparticles obtained from X-Ray films and different cap-ping agents. 108

Figure 4-98 (a, b)

XRD patterns of silver nanoparticles obtained from X-Ray films using NaOH and onion extract and garlic extract respectively. 109


TEM images of silver nanoparticles obtained from X-Ray films using NaOH and garlic extract. 110


TEM images of silver nanoparticles obtained from X-Ray films using NaOH and 1.25% (v/v) concentration of onion extract. 110

Figure 4-101 TEM image of silver nanoparticle obtained from X-Ray films using NaOH and black tea leaves extract. 111

Figure 4-102 UV-visible graphs of silver nanoparticles obtained from X-Ray films using NaOH and different capping agents. 111

Figure 4-103 Colour of the colloidal silver at different temperatures. 113




XRD patterns of silver nanoparticles synthe-sized in method V at different temperatures. 114

Figure 4-105 (a, b)

TEM pictures of as-synthesized silver nanopar-ticles of size 20–80 nm obtained from method V at 30°C. 114


TEM pictures of silver nanoparticles of size 15–25 nm synthesized using method V at 50°C. 115

Figure 4-107 (a–d)

TEM pictures of silver nanoparticles of size 5–65 nm as-synthesized by the method V at 70°C. 115

Figure 4-108 UV-visible graphs of silver nanoparticles corresponding to method V at different temperatures. 116

Figure 4-109 Colours of the colloidal silver for different strengths of precursor. 117


XRD graphs of silver nanoparticles synthesized using method V for different strength of precur-sor solution 0.005N, 0.01N, 0.02N respectively. 118


TEM pictures of silver nanoparticles of size 5–30 nm synthesized using method V for 0.005N strengths of precursor. 119


TEM images of silver nanoparticles synthesized using method V with 0.01N of the precursor solution. 119

Figure 4-113(a, b)

TEM pictures of as-synthesized silver nanopar-ticles of size 20–80 nm obtained using method V with 0.02N precursor strength. 119

Figure 4-114 UV-visible graphs for silver nanoparticles syn-thesized using method V and varying strength of precursor solution. 120

Figure 4-115 Colour of the solutions containing silver nanoparticles. 121


List of Figures 161

Figure 4-116 XRD graphs of as-synthesized silver nanopar-ticles obtained from method V with varying concentration of reducing and capping agent 1.25%, 2.5% and 6.25% (v/v) respectively. 121

Figure 4-117 TEM image of as-synthesized silver nanopar-ticles of size 30–80 nm obtained from method V for 1.25% (v/v) onion extract. 122

Figure 4-118(a, b)

TEM pictures of silver nanoparticles synthe-sized using method V with 2.5% (v/v) concen-tration of onion extract. 123

Figure 4-119(a, b)

TEM pictures of silver nanoparticles synthe-sized using method V with 6.25% (v/v) concen-tration of onion extract. 123

Figure 4-120 UV-visible graphs of silver nanoparticles syn-thesized using method V for 1.25%, 2.5%, and 6.25% (v/v) concentration of onion extract. 124

Figure 4-121 Colour of the colloidal silver due to different capping agents. 125


XRD graphs of silver nanoparticles synthesized using method V using onion, garlic and black tea leaves respectively. 125


TEM pictures of silver nanoparticles obtained from method V using 1.25% (v/v) concentra-tion of garlic extract. 126

Figure 4-124 TEM image of silver nanoparticles synthesized using method V with 1.25% (v/v) concentration of onion extract. 126


TEM images of silver nanoparticles synthesized using method V using 1.25% (v/v) concentra-tion of black tea leaves extract. 127

Figure 4-126 UV-visible graphs of silver nanoparticles syn-thesized using method V using 1.25% (v/v) onion, garlic and black tea extracts. 127


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List of tables

Table 1-1 Surface to volume ratio of nano spheres. 3Table 4-1 Effect of current on the size of nanoparticles and

UV-visible peaks for set 1, method I. 38Table 4-2 Effect of concentration of capping agent on the

size of nanoparticles and UV-visible peaks for set 2, method I. 42

Table 4-3 Sample and their combination sent to the lab for test against bacteria for 15 minutes. 43

Table 4-4 IC 50 values of the sample for cancer cell lines. 49Table 4-5 Effect of strength of precursor (silver nitrate) on

the size of nanoparticles and UV-visible peaks for set 3, method I. 54

Table 4-6 Effect of concentration of the capping agent on the size of nanoparticles and UV-visible peaks for set 4, method I. 58

Table 4-7 IC 50 values of the sample for cancer cell lines. 60Table 4-8 Effect of three different capping agents on the size

of nanoparticles and UV-visible peaks for set 5, method I. 64

Table 4-9 Effect of concentration of onion extract on the size of nanoparticles and UV-visible peaks for set 6, method I. 67

Table 4-10 1 ml sample + 0.1 ml culture [kept for 15 minutes] total solution plated on soya bean casein digest agar incubated at 37°C for 48 hrs. 68

Table 4-11 1 ml sample + 0.1 ml culture + ampicillin [10 mcg] [kept for 15 minutes] total solution plated on soya bean casein digest agar incubated at 37°C for 48 hrs. 69

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Table 4-12 1 ml sample + 0.1 ml culture + gentamicin [10 mcg] [kept for 15 minutes] total solution plated on soya bean casein digest agar incubated at 37°C for 48 hrs. 69

Table 4-13 1 ml sample + ampicillin [10 mcg] [kept for 15 minutes] and zone readings taken. 70

Table 4-14 1 ml sample + gentamicin [10 mcg] [kept for 15 minutes] and zone readings taken. 70

Table 4-15 Summary of the results for method I. 71Table 4-16 Effect of temperature of precursor solution on

the size of nanoparticles and UV-visible peaks for set I, method II. 78

Table 4-17 Effect of concentration of capping agent on the size of nanoparticles and UV-visible peaks for set 2, method II. 84

Table 4-18 Effect of three different capping agents on the size of nanoparticles and UV-visible peaks for set 3, method II. 87

Table 4-19 Summary of the results for method II. 88Table 4-20 Effect of concentration of the capping agent on

the size of nanoparticles and UV-visible peaks for set 3, method III. 99

Table 4-21 Effect of three different capping agents on the size of nanoparticles and UV-visible peaks for set 4, method III. 102

Table 4-22 Summary of the results for method III. 102Table 4-23 Effect of concentration of the capping agent on

the size of nanoparticles and UV-visible peaks for set 1, method IV. 108

Table 4-24 Effect of three different capping agents on the size of nanoparticles and UV-visible peaks for set 2, method IV. 112

Table 4-25 Summary of the results for method IV. 113

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List of Tables 165

Table 4-26 Effect of temperature on the size of nanoparticles and UV-visible peaks for set 1, method V. 117

Table 4-27 Effect of strength of precursor (silver nitrate) on the size of nanoparticles and UV-visible peaks for set 2, method V. 120

Table 4-28 Effect of concentration of capping/reducing agent on the size of nanoparticles and UV-visible peaks for set 3, method V. 123

Table 4-29 Effect of capping/reducing agents on the size of nanoparticles and UV-visible peaks for set 4, method V. 125

Table 4-30 Summary of the results for method V. 126

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Authors biographyDr. M. M. Malik was born in Bhopal, India in 1965. He received B.Sc. (1985) and M.Sc. (1988) degree from Barkatullah University, Bhopal, India. He was awarded PhD degree in 1992 by Jamia Millia Islamia (Central University) New Delhi, India on the topic “Electrical and structural studies of Chalcogenide Glasses.”

He was awarded Senior Research Fellowship by CSIR, New Delhi, in 1991. He visited ICTP Trieste

Italy in 1994 for research work. He joined Maulana Azad National Institute of Technology (MANIT) Bhopal, in July 1994 and currently is a professor and Head of the Department of Physics, Chairman, Nanoscience and Engineering Centre and Head, Department of Biological Sciences, MANIT. He has to his credit 68 publications in International Journals, 6 publications in National Journals, 22 pro-ceedings of International Conferences/Seminars and 12 Proceedings of National Conferences/Seminars. He has guided 13 PhD students. He is presently working on amorphous semiconductors, luminescence, electrets, green synthesis and nanomaterials. Dr. Malik is member of various professional societies which includes ISTE and Indian Chapter of ICTP Trieste, Italy.

Dr. Shweta Rajawat is presently a post-doctoral fellow in department of Physics, Maulana Azad National Institute of Technology (MANIT), Bhopal (India) under a scheme of University Grants Commission of Government of India. The research area is “Targeted drug delivery for cancer treatment using carbohydrate functionalized silver nanoparticles”. She obtained her PhD Degree at MANIT, Bhopal (India) on “Green synthesis and characterization of silver nanoparticles”.

Master’s degree and Bachelor’s degree in Science were awarded by Mohanlal Sukhadia University, Udaipur (Rajasthan, India) with certifi-cate of merit in each year of Bachelor Degree Program. Dr. Rajawat has also obtained Master’s Degree in Computer Application (MCA) from Indira Gandhi Open University, New Delhi (India).

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References...... 2004, 275, pp. 177–182. [4] Priyabrata Mukherjee ... The 21 volume Academic American ... ASME_19th Bionano_References.indd 136 Manila Typesetting Company 02/20/2018