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Biosynthesis, Characterization of Silver Nanoparticles Using Calotropis gigantea and

Screening their Free Radical Scavenging and Antibacterial Properties


A Synopsis Report
Submitted to
Department of Biotechnology
DCRUST, Murthal

In partial fulfilment of the degree of


Master of Technology

Under Supervision of :

Submitted by :

Dr. Reeti Chaudhary

Nitish kumar

Department of Biotechnology

14001509009

CONTENTS
INTRODUCTION
OBJECTIVES
REVIEW OF LITERATURE
MATERIALS AND METHODS
i) CHARACTERIZATION
ii) ANTIBACTERIAL ACTIVITY
iii) FREE RADICAL SCAVENGING ACTIVITY
SIGNIFICANCE OF STUDY
REFERENCES

INTRODUCTION

Nanotechnology is the art and science of manipulating matter at the nanoscale


(upto 100nm) to create new and unique materials and products. These
particles can be spherical, tubular, or irregularly shaped and can exist in fused,
aggregated or agglomerated forms.

Nanotechnology is often referred to as a general-purpose technology. Thats


because in its mature form it will have significant impact on almost all industries
and all areas of society.

It offers better built, longer lasting, cleaner, safer and smarter products for the
home, for communications, for medicine, for transportation, for agriculture, and
for industry in general.

Silve nanoparticles are of great interest for use as antimicrobial agents. Smaller
Silver Nanoparticles have greater antibacterial activity hence they are new
generation of antimicrobials

Nanotechnology silver particles have great potential in the medical field


including diagnosis, medical device coating, and health care.

Biological systems can be used for the synthesis of silver nanoparticles

Silver nanoparticles are known as excellent antimicrobial agents, and


therefore these can be used as alternative disinfectant agents in drinking
water.

silver nanoparticles have been synthesized using medicinal plants, e.g.


Calotropis gigantea, Aloe vera etc.

Calotropis is used alone and sometimes with other plants to cure variety
of human and animals ailments. It is used in various polyherbal
preparations. Different parts of the plant have immense potential to cure
various diseases and disorders.

Used to treat common disease such as fevers, rheumatism, indigestion,


cough, cold, asthma, elephantiasis, vomiting, diarrhea etc.

Medicinal properties of Calotropis gigantea


Antimicrobial activity
Asthma

Skin diseases
Antiviral activity

Anti-inflammatory activity
Analgesic and Antinociceptive
Activities

Anti cancer activity

Activity
Arrow poison

PartsAnti
Usedtumor activity
Latex

Biocidal activity

Latex

Brewing and to curdle milk

The bark and


latex

Cleansing water

Leaves and its


Saps

Energy plantation
Leather tanning
Fungicidal, insecticidal

Whole Plant
Whole Plant
Whole Plant

Cold & Fever

OBJECTIVES

To synthesize silver/gold nanoparticles using


Calotropis gigantea.

Characterization of synthesized nanoparticles


using X-Ray Diffraction, UV-Vis spectroscopy,
FTIR and SEM/TEM.

To screen out antimicrobial and free radical


scavenging

activity

nanoparticles.

of

synthesized

silver

REVIEW OF LITERATURE
Many studies have been conducted on, silver nanoparticles using different
parts of plants like nodal, leaves and whole plant by various researchers
S.No
1

Author & year


Work done
M. Gondwal et al, biological evaluation
2013

green

synthesis

of

Outcomes
and Face centered cubic (FCC) structure and
silver size was ranging from 19 to 45 nm (L) and

nanoparticles using aqueous 26 to 38 nm (S) respectively, spherical


extract of Calotropis procera

shape, significant antimicrobial potential


against K. pneumonia and S. typhi,

V. Baskaralingam et Green synthesis of Silver A decrease in colonies was observed


al, 2012

nanoparticles
Calotropis

through at 5 mg/mL of silver nanoparticle


gigantea leaf

extracts and evaluation of


antibacterial activity against
Vibrio alginolyticus

concentration

and

the

complete

inhibition of V. alginolyticus was


observed at 20 mg/mL of silver
nanoparticle concentration on LB
agar plates

S.No

Author & year

J. Sivakumar et al, Biosynthesis


2011

Work done

Outcomes

of

Silver nanoparticles

Nanoparticles
Using Calotropis gigantean
Leaf

was

observed

with different time intervals,


the colour of the reaction
medium changed rapidly from
colorless to brown.

Singhal et al, 2011 Biosynthesis


antimicrobial
silver

and Silver nanoparticles size 430 nm,


activity

nanoparticles

Ocimum sanctum.

of within 8 min of reaction time,


using exhibit more antimicrobial activity
on gram-negative microorganism
than gram-positive .

Mohapatra et al, Citrus


2010

limon

extract size range 10-30 nm, addition of

mediated synthesis of silver NaOH is a key factor for rapid


nanoparticles

biosynthesis, alkaline environment


is favorable

S.No

Author & year

G. Geoprincy et
al, 2012

Work done

Outcomes

green synthesis of silver Reduction of silver ions to silver


nanoparticles

nanoparticles, UV-Vis spectrum,


scanning electron microscope, X
ray

diffraction

and

transfer infra-red spectra.

Fourier

SYNTHESIS OF SILVER
NANOPARTICLES
Collect fresh leaves
Wash with distilled water to remove dust (3 times).
Finely chopped + 100ml D.W
Boil for 15 min. and then cool (at 60 C ) .
Filter using whatmann filter paper.
Freeze dry the extract (-70C2C; 48 h)
Store at 4C
Continued

CONTINUED

Mix extract with 1mM AgNO3 aqueous solution.


Keep at room temperature for 24 hours.
Observe color change (yellowish - brown).
Observe synthesis using UV-Visible spectroscopy.

CHARACTERIZATION

Characterization
refers to the study of materials
features such as its composition, structure, & various
properties like physical, electrical, magnetic etc.
UV visible spectroscopy: In general 200-800 nm light wavelength
is used for the characterization of size range 2-100 nm.
X-Ray Diffraction (to study structure, crystallinity, particle size,
distribution, particle shape and homogeneity of whole sample)

FTIR (to detect functional groups responsible for bioreduction)

SEM/ TEM (size/morphology).

ANTIBACTERIAL ACTIVITY

Grow bacterial test organisms in LB broth for 24 hours.


Spread LB broth culture over Muller Hinton Agar using
cotton swab/ spreader.
Dry for 10 minutes.
Put the 4 discs with following solutions as shown in figure
& compare zones of inhibition.
Calotropis
gigantea
Antibiotic

AgNPs
AgNO3

ABTS RADICAL SCAVENGING ASSAY


Incubate ABTS (7mM) & Potassium persulfate (2.45mM)
mixture at room temperature in dark for 16 h.
Test sample (10ml) + Distilled water (50 ml) + ABTS (140ml)
Record absorbance at 734nm.
Use(6-hydroxy-2, 5, 7, 8-tetramethylchroman-2- carboxylic
acid ) as positive control.
Calculate % Inhibition as:
% Inhibition = [A (control) A (sample)]/ A(control) 100.

DPPH RADICAL ASSAY


Add AgNps (equal volume) to 0.1mM methanolic DPPH solution.
Incubate reaction mixture for 30 min at room temperature under
shaking conditions.
Record absorbance at 517nm.
Use(6-hydroxy-2, 5, 7, 8-tetramethylchroman-2- carboxylic acid )
as positive control.
Calculate % Inhibition as:
% Inhibition = [A (control) A (sample)]/ A(control) 100.

REFERENCES

Gamble J S; Flora of the Presidency of Madras, Vol. I, II, III, Botanical survey of India,
Calcutta, 1935.

Saikia D, Gogoi PK, Phukan P, Bhuyan N, Borchetia S, Saikia J. Green synthesis of silver
nanoparticles using Asiatic Pennywort and Bryophyllum leaves extract and their antimicrobial
activity. Adv. Mater. Lett. 2015; 6: 260-264.

manjul gondwal and geeta joshi nee pant* biological evaluation and green synthesis of silver
nanoparticles using aqueous extract of calotropis procera Int J Pharm Bio Sci 2013 Oct; 4(4):
(P) 635 643

Baskaralingam Vaseeharan,1 Clara Gunapoorani Sargunar,2 Yong Chin Lin,3 Jiann Chu Chen3
Green synthesis of Silver nanoparticles through Calotropis gigantea leaf extracts and
evaluation of antibacterial activity against Vibrio alginolyticus Italy Nanotechnology
Development 2012; 2:e3 doi:10.4081/nd.2012.e3

G.geoprincy1, b.n.vidhya srri2, u.poonguzhali2, n.nagendra gandhi1, s.renganathan, a review


on green synthesis of silver nanoparticles Asian J Pharm Clin Res, Vol 6, Suppl 1, 2013, 8-12

Singhal G, Bhavesh R, Kasariya K, Sharma AR, Singh RP. Biosynthesis of silver nanoparticles
using Ocimum sanctum (Tulsi) leaf extract and screening its antimicrobial activity. J Nanopart
Res. 2011; 13: 2981 2988.

. J. Sivakumar, C. Premkumar, P. Santhanam and N. Saraswathi Biosynthesis of Silver


anoparticles Using Calotropis gigantean Leaf African J. Basic & Appl. Sci., 3 (6): 265-270,
2011

Farooqui MA, Chauhan PS, Krishnamoorthy P, Shaik J. Extraction of silver nanoparticles from
the leaf extracts of Clerodendrum inerme. Dig J Nanomater Bios. 2010; 5: 4349.

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