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1.

1 Factors affecting biosynthesis of nanoparticles


In previous study, it was noticed that several properties such as the size, shapes, optical
properties and amount of the synthesized nanoparticles can be greatly affected by several
factors. Distinction of shapes and size of the previously synthesized nanoparticles were
reported to occur due to variation of process parameter such as plant broth concentration,
mixing ratio of plant extract to metal precursor (i.e. AgNO
3
) solution, interaction time
and pH of the solution (Tripathy et al., 2009).

1.1.1 Plant extract concentration and mixing ratio of plant extract to metal precursor
The plant extract concentration can lead to different characteristic in UV-vis spectra of
the nanoparticles. In a study conducted by Tripathy et al., (2009), three concentration
(5%, 20%, and 40%) of Neem broth were tested. It was then observed that particles
synthesized with 5% broth concentration gave a very weak Plasmon resonance band
(absorbance value 0.509) at 417 nm. Then, as the concentration increased to 20%,
max

increased to 444 nm with absorbance 0.815. However, further increase in concentration
to 40%
max
change slightly to 441 nm with decrease in absorbance to 0.476. The
variation of
max
was corresponded to changes in particle size due to change in
concentration. Consequently, the ratio of silver ions to capping agent/ stabilizing agent
influence the size of synthesized nanoparticles.

Next, another factor that is closely related to plant extract concentration is, ratio of the
extract to selected metal precursor. In the same study, there are several ratios that have
been tested and the maximum absorbance was noted for 20% concentration at 1:4 Neem
extract to AgNO
3
mixing ratio. Another study also found that reduction reaction occurs
rapidly with increase in addition of extract and the colloid obtained in 30 min (Philip,
2010).

Philip (2010) reported that Au nanoparticles of different size and shape were synthesized
using hibiscus leaf extract by varying the ratio of metal salt and the extract. From the
prepared colloid solution with (varying the addition of plant extract; 10, 20, and 30 ml),
variation of morphology and size of synthesized nanoparticles were also obtained. The
observation showed that colloid containing 10 ml of plant extract produces large number
of anisotropic particle in the company of smaller particle being multi-branched. In the
colloid with addition of 20 ml plant extract, nanoparticles with triangular, hexagonal,
dodecahedral and triangular shapes are seen. With the maximum addition of 30 ml, the
particles produced are almost spherical with size of ~14 nm.

Hence, from the observation, it was noticed that the size and propensity of triangular
particles decrease with increase addition of plant extract. The particles tend to become
dodecahedral and spherical. From this study, it was learned that even if lower quantities
of the extract sufficient of the reduction of chloroaurate ions, most of the quasi-spherical
nanoparticles was not prevented from aggregating due to deficiency of biomolecules to
act as protecting agent.

Biosynthesis of gold and silver nanoparticles mediated by cashew leaf powder that was
previously done by Sheny, Mathew and Daizy, (2011), claim that lower amount of plant
material were sufficient to bring about reduction. It was also reported that for generation
of small and stable Au nanoparticles from the dried leaf powder, 12 mg is sufficient. For
amount below than that, the UV-vis absorbance decrease. The Uv-vis spectra of Ag
nanoparticles show a linear relationship between absorbance and quantity of extract. A
lower quantity of 5 mg of leaf powder gave absorption in visible region which will be
more suitable than their higher quantities.

1.1.2 Reduction temperature
Temperature at which reduction of metal ion occurred also played an important role in
determining the effectiveness of the synthesis. In a study conducted, reaction occurred at
300 K and 373 K was compared and it was found that at higher temperature, even 0.1 ml
of extract was sufficient to bring about reduction of Au
3+
. Comparatively, creating stable
colloid solution in lower temperature medium required more plant extract (2.5 ml)
compared as in higher temperature (0.6 ml). Reduction occurs at high temperature
leading to large-size and more stable Au nanoparticles. For synthesizing Ag
nanoparticles at 300 K, 1 ml was required while at 373 K, 0.2 ml was required. Ag
nanoparticles synthesized at higher temperature showed higher stability. Hence from the
study (Sheny, Mathew and Daizy, 2011) it can be concluded that at higher temperature,
minimal amount of plant extract can initiate metallic reduction and the particles
produced were large with better stability.



1.1.3 Interaction time
By referring to test conducted by Tripathy et al., 2009, they found that in short
interaction time of 4 h, nanoparticles below 20 nm with nearly spherical shape was
produced. They also claim that the aggregation and shape anisotropy increase with
increasing interaction time.
1.1.4 The solution pH
In research conducted by Sheny, Mathew and Daizy, (2011); observed a sharp and
symmetric peak in UV-vis spectrum of Au nanoparticles at pH of 6 and a broad peak
(indicates large sized particles) at pH of 4. Hence, the research concluded that Au
nanoparticles stable at pH of 6. The study also reported for Ag particles, their absorbance
increases with pH, where at lower pH peak broadening in occurred as an attribute to
large particle size.

In alkaline pH range, stability of cluster distribution was enhanced while the tendency
for aggregation of the particles decreases (Tripathy et al., 2009). This is supported by a
large fall in the flocculation parameter in alkaline pH ranges that has been reported by
Sastry et al., (1997), which might have caused decrease in aggregation. Hence, in the
study conducted by Tripathy et al., 2009, pH 2 of Ag solution failed to be synthesized as
nanoparticles and flocculation was clearly observed in the medium. They reported that
the stability of nanoparticles in the presence of additives has been found to be a function
of solution pH since OH
-
can change the surface charge on the nanoparticles. Therefore
at alkaline the stability of the cluster distribution was enhanced primarily due to
complete charging of the cluster. As a result the repulsive electrostatic/electrosteric
interaction was maximized. This enhanced stability and decreased aggregation.
















One of prominent factor that need to be consider during synthesizing nanoparticles is
temperature. According to Armendariz et al., temperature variation in reaction condition
may lead to fine tuning of the shape, size and optical properties of the nanoparticles
synthesized. In research conducted by Song, Jang, and Kim (2009) more than 90% of
leaf extract of two plants Magnolia kobus and Diopyros kaki was converted to gold
nanoparticles at a reaction temperature of 95
o
C in few minutes, suggesting reaction rates
higher or equivalent to synthesizing it chemically. It was also observed that the size of
gold nanoparticles synthesized by Muralidharan et al., increased at higher reaction
temperature. This claim was due to at higher reaction temperature the fusion efficiency
of micelles which dissipates super-saturation also increase.

The size of nanoparticles synthesized were also greatly influence by pH of the medium.
This was proven in the case of green synthesis of gold nanoparticles mediated by Avena
Sativa extract (Shanker, Bhargrava and Sastry, 2005).

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