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Chapter 1
INTRODUCTION
Nanotechnology is an important field of modern research dealing with synthesis, strategy
and manipulation of particles structure ranging from approximately 1 to 100 nm in size. Among
all noble metal nanoparticles, silver nanoparticles (AgNP) gained interest because of its
antimicrobial properties and wide range of applications in different fields such as medicine,
electronics, food industry, catalysis, optics, in the environment, etc. In addition, silver
nanoparticles have high surface area to volume ratio, unique chemical and physical properties.
Currently the development of green method in the synthesis of AgNP has an advantage over the
conventional way of synthesizing AgNP which uses high energy, temperature, pressure and toxic
chemicals because it is cost effective and eco-friendly. Green route is suitable for the synthesis of
silver nanoparticles that will be used in medicine and health products because the particles
surface does not contain toxic reducing agents (Ahmed, Ahmad, Lal Swami, Ikram, 2015).
The presence of flavonoids and phenolic compounds in plants and weeds have been
found to give multiple biological effect. The use of weed as source of these phytochemical
constituents have not yet explored. The researchers focused on the study of this weed
(Phyllanthus niruri). Phytochemicals that are responsible for the reduction of Ag + ions into Ag0
and the stabilization of the AgNP is due to the combination of biomolecules such as proteins,
amino acids, enzymes, polysaccharides, alkaloids, tannins, phenolics, saponins, terpenoids and
vitamins which are already established in the plant extracts having medicinal values and are
environmental benign, yet chemically complex structures (Kulkarni and Muddapur et al., 2014).

Moreover, the rate of nanoparticles synthesis is fast and the reducing form is more stable in plant
extracts (Wonsawat, 2014). Phyllanthus niruri is locally known as sampa- sampalukan and
commonly regarded as roadside and garden weed which is endemic in the Philippines and has
been reported to have hepatoprotective effect, inhibiting HIV replication, lipid lowering activity,
antidiabetic activity, anti-malarial activity, anti-spasmodic activity, analgesic activity, antioxidant
activity and inhibiting chromosomal aberrations (Montejo, Mondonedo, Lee, Ples, Vitor II,
2015).
Since P. niruri is abundant and regarded as a garden weed in the Philippines the
researchers were inspired to use it as reducing and stabilizing agent in the synthesis of silver
nanoparticles because of its rich phytochemical constituents. There has been no studies recorded
yet on this weed, so the researchers subjected the extracts to a microwave oven synthesis of
silver nanoparticles using P. niruri weed in the Philippines.
1.1 Objectives
This study aimed to synthesize silver nanoparticles via bioreduction using leaf extracts of
Phyllanthus niruri. Specifically the study attempted to:
1. determine the:
1.1 phytochemical constituents of the leaf extract of Phyllanthus niruri such as;
1.1.1 flavonoids
1.1.2 tannins
1.1.3 terpenoids
1.1.4 reducing sugar
1.2 effect of microwave irradiation time in the reduction of silver ions into
silver nanoparticles by UV-Vis Spectroscopy.
2. characterize the synthesized silver nanoparticles in terms of:
2.1 stabilizing molecules on the surface through FTIR

2.2 morphological feature and elemental composition through SEM-EDX

3. compare the antibacterial activity of ethanolic leaf extract of Phyllanthus niruri and
synthesized silver nanoparticles against:
3.1 Staphylococcus aureus
3.2 Escherichia coli
3.3 Haemophilus influenzae
3.4 Streptococcus pneumoniae
1.2 Significance of the Study
The results of this study will determine the efficacy of synthesized silver nanoparticles
using Phyllanthus niruri leaf extract as a potential anti-microbial agent. The comparison of the
biological activity of the synthesized silver nanoparticles from Phyllanthus niruri and the
aqueous leaf extract lead to further study on other therapeutic application of silver nanoparticles.
Plant extract mediated synthesis of silver nanoparticles is significant because it will not involve
the use of hazardous chemicals and high energy consumption. Search and utilization of
environment-friendly, rapid and cost effective synthesis of silver nanoparticles would be
significant.
1.3 Scope and Limitations
The study is focused on the synthesis and characterization of silver nanoparticles via
bioreduction of silver ions using ethanolic leaf extract of Phyllanthus niruri. The Phyllanthus
niruri leaves was collected from the road sides of San Pedro, Laguna. Green color and
undamaged leaves was used in this study.

The extraction, phytochemical screening and synthesis of silver nanoparticles was

conducted at the chemistry laboratory of Technological University of the Philippines Manila.
Domestic microwave oven was used in the synthesis. The reduction of Ag + to Ag0 was monitored
using Thermo-Scientific Genesys 10S at College of Pharmacy, University of the Philippines
Manila. Determination of stabilizing molecules on the surface of silver nanoparticles was
determined using Thermo-Scientific Nicolet 6700 FTIR at De La Salle University.
Morphological features and elemental composition was analyzed using JEOL JSM-5310
Scanning Electron Microscope equipped with Energy Dispersive X-ray at De La Salle University
Manila.
Antibacterial activity of ethanolic extract of Phyllanthus niruri and ethanolic extract of
Phyllanthus niruri containing nanoparticles will be compared on gram-negative (E. coli), grampositive (S. aureus), and on fastidious gram-negative(H. influenzae) and gram-positive (S.
pneumoniae) In-vitro antimicrobial test by disc diffusion method was conducted at Department
of Health-Research Institute for Tropical Medicine (DOH-RITM), Microbiology Department
using standard procedures described by Clinical and Laboratory Standards Institute.
Synthesis procedures were adapted from the work of Pal, Deb, Deshmukh (2013).