Research Synopsis

Green synthesis of Silver Nanoparticles from Euphorbia grantii

Oliv. Leaf extract: Their structural characterization and
biomedical application

M.Phil. Botany

Session: 2022-2024

Submitted By: Mr.Hilal Ahmed

Supervisor: Dr. Faizan Ullah

Department of Biotechnology, University of Science and Technology,

Bannu, KPK, Pakistan

 1. INTRODUCTION

Nanotechnology

Nanotechnology is a flourishing science that deals with usage and production of nanosized
particles. It can be best explained as the science that involves synthesis, design, application
and characterization of materials whose smallest possible length in one dimension is at nano
meter scale (Gavarkar et al., 2014). Nanotechnology is currently on the rise and will have
everlasting impact on humanity by resolving the basic issues which faces humanity in the
context of health and treatment of disease. And these remarkable attributes of nanoparticles is
due to their practical application in areas like medicine (Pauwels et al., 2008) chemical
detection, electronics and catalysis (Hall et al., 2011). Nanotechnology develop materials,
characterize it which are at nanoscale and thus in this way open a vast area for application
(Jeremy et al., 2009). Advancement in nanotechnology introducing new metal nanoparticles
is opening new era of vast application of nanoparticles and creating research opportunities
(Sharma et al., 2009).

Nanoparticles

Nanoparticles have size of 1 to 100 nanometre, and possess unique physical and chemical
properties. It because of their large surface area compared to volume and having smaller
particle size (Kato et al., 2011). The application of nanoparticles is huge and in this regard
nanoscience and nanotechnology is rapidly developing. Nanoparticles of silver, gold, zinc,
and iron has been used in medicines, agriculture and other industries.

Different ways for nanoparticles synthesis

Several methods can be employed for nanoparticle synthesis as briefly described below.

1 Physical process:

The most important physical approaches are Evaporation – condensation and Laser ablation.
Silver, gold lead sulphide and Zinc oxide were synthesized by the method of evaporation-
condensation method (Kruis et al., 2000).

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2 Chemical synthesis:

Chemical reduction with the help of reducing inorganic and organic compounds is for
chemical synthesis of silver nanoparticles. Different compounds like sodium Borohydride,
sodium citrate, ascorbate, and nascent hydrogen, Tollens reagent, N, N-dimethylformamide,
compounds containing polyols and poly ethylene glycol are used as reducers. These reducers
reduce the silver ion and as a result of reduction metallic silver is formed (Evanoff et al.,
2004).

Biological synthesis:

Silver Nanoparticles can be synthesized by using biological methods in which toxic and
expensive chemicals are not used (Ahmad et al., 2003). The bio molecules found in the
extract of certain organism like enzymes, proteins, polysaccharides etc which reduces the
metal ions are environmentally safe (Korbekandi et al., 2009).

1 By using bacteria:

Use of bacteria as an agent for nanoparticles synthesis was reported by using pseudomonas
stutzeri AG259. Widely accepted mechanism for silver biosynthesis is the presence of
enzyme nitrate reductase. Nitrate is converted into nitrite by this enzyme. This enzyme plays
a vital role in synthesis of silver nanoparticles (Haefeli et al., 1984).

2 By Using Fungi:

Nanoparticles are synthesized by Fungi in large amount because they have large quantities of
secretions of proteins. This directly links to the nanoparticles synthesis in increased amount
(Mohanpuria et al., 2008). In F. Oxysporum it is supposed that the NADPH – dependent
nitrate reductase and extra cellular quinine shuttle process are responsible for synthesis of
nanoparticles (Ahmad et al., 2003).

3 Use of plant as a mean for nanoparticles synthesis:

Significance of using plant extract for silver nanoparticles synthesis is that plants are easily
available, safe to work with and nontoxic in nature. Phytochemicals like terpenoids,
aldehydes, amides, carboxylic acid, flavones, and ketones are involved in plant mediated
silver nanoparticles synthesis (Jha et al., 2009).

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Need for biosynthesis of nanoparticles:

As physical and chemical process are economically costly and toxic to the environment so
there is a need for the green synthesis of nanoparticles. Chemical methods produce toxic
chemicals absorbed on the surface which can show lethal effects when they are used in
medical treatment and procedures. Green synthesis is preferred over physical and chemical
methods because it is less costly, environmental friendly, easily scaled up for mass
production, no need of high pressure, energy and temperature (Parashar et al., 2009).

Silver nanoparticles

It is reported that nearly 25% of all nanotechnology consumer products involve AgNPs
(Vance et al., 2015). Because of their well-known antibacterial and antifungal properties,
they can be used in household products, food packaging, textiles, medical devices, antiseptics
in healthcare delivery, and personal healthcare (Ahamed et al., 2010; Quang et al., 2019).
AgNPs can also be used in electronic devices and wastewater treatment because of their good
electrical conductivity and photochemical properties (Quang et al., 2019; Zhang et al., 2016).
In the agriculture sector, AgNPs were developed as plant-growth stimulators (Steinitz &
Bilavendran 2011; Monica & Cremonini, 2019), fungicides to prevent fungal diseases (Alavi
& Dehpour 2010), or agents to enhance fruit ripening (Sah et al., 2011; Vinkovi´c et al.,
2017). The growing consumption of AgNPs inevitably increases the chance of release into
the environment during AgNPs synthesis and incorporation into products, as well as handling
and recycling or disposal of these products (Hedberg et al., 2014; Lombi et al., 2014). AgNPs
are expected to flow into environment as surface waters (Sun et al., 2014), and the main
pathway is through biosolids from wastewater treatment (Gottschalk et al., 2009; Mueller &
Nowack 2008). Indeed, AgNPs are detected widely in water and soil; they accumulate in the
soil or water reservoirs in large quantities (Benn and Westerhoff 2008; Gottschalk and
Nowack 2011). An analysis of the wastewater from a sewage treatment plant indicated
existence of AgNPs with a size of 9.3 nm and a concentration of 1900 ng/L (Hoque et al.,
2012).

Euphorbia grantii Oliv.

The genus Euphorbia (family Euphorbiaceae) has been reported to have some promising
activities as anti-inflammatory, anticancer, immunoregulation, and fibrinolytic actions
(Campos et al., 2016).Phytochemical investigation of the family Euphorbiaceae showed the

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presence of saponins, diterpenes, triterpenes, flavonoids, lectins, phorbol esters, and
glycoproteins.( Balakrishnan & Chakrabarty 2007). Euphorbia grantii Oliv was reported for
its antiviral activity against several reported viruses (Van Hoof et al., 1984).Triterpenes are
biologically active metabolites classified into tetracyclic and pentacyclic triterpenes and have
recently caught broad attention. They have been reported as anti-inflammatory, antiviral,
antibacterial, sedative, immunoregulatory, blood sugar regulatory, blood pressure-lowering,
and antitumor agents (Zhang et al., 2014).Recent reports indicated that triterpenes could
directly inhibit the tumor growth, both in vivo and in vitro via several mechanisms(Abbas et
al., 2013; Elkhayat 2014).

Statement of the problem

Silver has been reported to possess antimicrobial effects with particular qualities of
conductivity, stability, and activity (Das et al., 2011). According to Quang et al., the
antibacterial action of silver and its compounds has been long known as an antibiotic to treat
several infectious disorders and burn wounds. Researchers are being challenged to discover
novel and effective antimicrobial reagents that are both resistant and inexpensive.

Research hypothesis

Euphorbia grantii Oliv has been reported for antioxidant potential. Therefore it is
hypothesized that plant extract of E.grantii Oliv would be highly effective in the synthesis of
silver nanoparticles. Silver is a metal with large scale industrial applications. It is used in
many pharmaceutical products. It is known that nanoparticles have large surface area and are
more effective than bulk materials. It is also hypothesized that pharmacological significance
of silver nanoparticles will be higher than bulk.

Research aims and Objectives

Aim of study is to synthesize silver nanoparticles from Euphorbia grantii plant extract. The
following objectives will be achieved.

Characterization of the silver nanoparticles by UV, SEM, FTIR, XRD.

To evaluate AgNPs for various in vitro pharmacological activities such as antimicrobial,

antioxidant, phytotoxic and cytotoxic properties.

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 SCOPE AND SIGNIFICANE

Need for biosynthesis of nanoparticles rose as the physical and chemical processes were
costly. Often chemical synthesis method leads to presence of some of the toxic chemical.
These chemical absorbed on the surface of nanoparticle and may have adverse effect in the
medical applications. Green synthesis is preferred over chemical and physical method
because it is

 Cost effective
 Environment friendly
 Easily scaled up for large scale synthesis and in this method there is no need to use
high pressure, energy, temperature and toxic chemicals.
 Opening new era of synthesizing drugs at nano scale which can cure current
challenging disease like cancer.

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 Materials and Methods

Plant Extract Preparation:

Fresh and healthy Euphorbia grantii Oliv.plant will be collected, washed, cleaned, and then
dried at room temprature. After fully drying, the plant will be cut in pieces and make their
powder by grinder or pastal motor. Then we will weigh the powder and add 100mL deionized
water will added to the plant powder. Then kept in closed container for 7 days, after this
process the mixture will be filtered by whatman filter paper, we will have filtered aqueous
plant extract in beaker, now we will put the mixture in open container to evaporate the water
and crude extract will be collected in tubes for further experiments.
Synthesis of Nanoparticles
Method for nanoparticles synthesis will be used according to standard protocol of Khan et
al., (2018) .

Characterization of nanoparticles

Silver nanoparticles will be characterized through the following techniques.

1. UV spectroscopy
2. FTIR
3. XRD
4. SEM

Biological evaluations

Antibacterial Activity

The bacterial activity of AgNPs will be assessed using the usual approach (disk diffusion
method) described by Revati et al., (2013).

Antifungal Activity

The silver nanoparticles will be tested for antifungal activity using method of Jogee et al.,
2017.

Insecticidal activity

The insecticidal activity will be determined by the method of Maqsood et al., 2020.

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Antioxidant activity

To evaluate the antioxidant potentiality of AgNPs and plant extract the modified technique
will be used (Williams et al., 1995).

Phytotoxicity

The phytotoxic potential of the synthesized silver nanoparticles in different concentrations

i.e. 25 – 200 mg/L will be determined on maize plant. The effects of the nanoparticles on
seed germination, seedling growth, physiological and biochemical traits will be determined.

Photosynthetic pigmentation

Determination of photosynthetic pigments will be done by Kirk (1968) method.

Phenolic contents of plants

Phenolic contents of plants will be find out through Adom et al., 2005.

Antioxidant activity

Antioxidant activity of plants will be carried out by Blois (1958) method.

Heavy metal detection activity

The heavy metal detection activity will be carried out by the method of Abul kalam et al.,
2018.

Tentative schedule/Timeline:

The whole research shall be carried out in one year and shall be divided into three phases;

every phase will be of four months.

Phase 1 Collection of plant materials, extraction, nanoparticles synthesis

Phase 2 Biological evaluation of nanoparticles, characterization

Phase 3 Thesis write up and research paper publication

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