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Materials Research Bulletin 85 (2017) 6473

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Materials Research Bulletin


journal homepage: www.elsevier.com/locate/matresbu

Green synthesis and characterization of cadmium sulphide


nanoparticles from Chlamydomonas reinhardtii and their application as
photocatalysts
M. Divya Rao* , Gautam Pennathur*
Centre for Biotechnology, Anna University, Chennai 600025, India

A R T I C L E I N F O

Article history:
Received 29 March 2016
Received in revised form 14 August 2016
Accepted 30 August 2016
Available online 31 August 2016
Keywords:
A. Semiconductors
C. Electron microscopy
C. X-ray diffraction
D. Optical properties
D. Catalytic properties

A B S T R A C T

This paper describes a facile, green method for the synthesis of cadmium sulphide (CdS) nanoparticles
from Chlamydomonas reinhardtii. Morphological analysis by electron microscopy revealed the presence of
spherical particles measuring approximately 5 nm. Structural analysis by powder X-ray diffraction and
Fourier transform infrared spectroscopy conrmed the presence of cubic CdS nanoparticles that were
capped with algal proteins. Optical analysis showed a signicant blue shift in the optical band gap that
could be ascribed to quantum connement. The photocatalytic ability of these nanoparticles against
methylene blue under UV light was studied & was found to degrade 90% of the dye within 90 min.
Trapping experiments indicate that photogenerated holes, OH were the main reactive species
responsible for dye degradation. This one-step strategy using algal cell free extract to produce CdS
nanoparticles is an economical, environmentally friendly approach for the large-scale synthesis of CdS
nanoparticles that can be used in photocatalysis.
2016 Elsevier Ltd. All rights reserved.

1. Introduction
Cadmium sulphide (CdS) belongs to the IIVI group of
semiconductor nanoparticles. Due to quantum connement, these
nanoparticles have unique optoelectronic properties that signicantly differ from the bulk material. They have been extensively
studied in photocatalysis [36], used as biological sensors [17] and
light emitting diodes [38]. The expansion of industries and
growing concern over environmental impact of organic dyes in
water bodies has led us to evaluate alternate strategies for water
purication. Semiconductor nanoparticles like CdS with their
unique photophysical and photochemical properties are ideal for
use in the degradation of organic dyes via photocatalysis, using UV
or visible light as a source of illumination [14]. Conventional
methods of fabrication include chemical methods, microwave
heating, hydrothermal approaches and precipitation from liquid
reactions [34]. These methods usually employ high temperatures
and extremely toxic reagents for prolonged periods of time; they
are also expensive and in many cases require specialized
equipment.

* Corresponding authors.
E-mail addresses: divyarau@gmail.com (M. D. Rao), pgautam@annauniv.edu
(G. Pennathur).
http://dx.doi.org/10.1016/j.materresbull.2016.08.049
0025-5408/ 2016 Elsevier Ltd. All rights reserved.

Organic materials such as mercaptoethanol, thiophenol and


thioacetamide are frequently used as capping agents in the
synthesis of cadmium sulphide nanoparticles. These chemicals are
extremely harmful and pose a signicant environmental hazard
[34]. Their potential in the eld of hydrogen storage and in the
degradation of toxic dyes via photocatalysis emphasizes the need
to develop safer strategies for the large-scale synthesis of these
nanoparticles. Biological materials offer an interesting solution,
they are environmentally safe, do not require harsh processing
conditions and are cheaper to use [31]. A number of biological
sources have been used as capping agents in the synthesis of CdS
nanoparticles; however, there is only one report that discusses its
synthesis by employing R-phycoerythrin, a pigment isolated from a
marine cyanobacteria. There have been no reports till date that
make use of algae for the synthesis of these nanoparticles. Fresh
water algae like Chlamydomonas reinhardtii are ideal candidates
because they are easy to culture and scale up, cost effective and do
not pose a threat to the environment [2,12].
One of the greatest challenges in the recent past has been
purication of water, as close to 4 billion people have little to no
access of clean water for daily consumption. Population boom,
vagaries in weather and rapid industrialization are some of the
factors responsible for growing water shortage [3]. Paper and
textile industries employ organic dyes that contaminate waste

M.D. Rao, G. Pennathur / Materials Research Bulletin 85 (2017) 6473

water; up to 15% of synthetic dyes are lost as waste water during


manufacturing and processing operations. These dyes pose
signicant threat to the environment as they limit the amount
of sunlight that can penetrate into the water bodies thus affecting
aquatic life. Further, some dyes even at low concentrations are
extremely toxic to aquatic life and are responsible for the
destruction of aquatic communities [39]. The extensive variability
in the nature of the dyes and the stability of their molecular
structures makes them recalcitrant to remediation by various
physicochemical and biological methods. Advanced oxidation
processes (AOP) are attractive alternatives since they produce
highly reactive transitory species that mineralize organic contaminants. Of these, photocatalysis is a safe and economical
method; moreover heterogeneous photocatalysts such as CdS have
shown efciency in remediation by mineralization of contaminants to biodegradable compounds [3].
This paper discusses a novel, rapid, green approach in the
synthesis of cadmium sulphide nanoparticles using the environmentally benign fresh water alga Chlamydomonas reinhardtii. We
believe that this method is advantageous as it avoids further
downstream processing for the extraction of the nanoparticles. The
structural and optical properties of these nanoparticles were
investigated using UVvis absorption spectroscopy, photoluminescence spectroscopy (PL), High resolution scanning electron
microscopy (HR-SEM), High resolution transmission electron
microscopy (HR-TEM), Powder X-ray diffraction (XRD) and
Dynamic light scattering (DLS). We have demonstrated that algal
biomolecules are involved in the reduction and stabilization of the
CdS nanoparticles using Fourier transform infrared spectroscopy
(FTIR) and have posited a possible mechanism that explains its
formation. The photocatalytic activity of these nanoparticles
against methylene blue an organic dye was extensively studied.
2. Materials and methods
2.1. Preparation of algal cell-free extract
The Culture collection Centre, CAS in Botany, University of
Madras, India provided Chlamydomonas reinhardtii. This is a
freshwater green alga that is abundantly found water bodies.
The algae were grown as a suspension in 1L Erlenmeyer asks
containing 225 ml of Bolds basal medium (BBM) with a twelvehour photoperiod at 25  C for seven days. The cells were harvested
by centrifuging at 12,000 rpm for 20 min at 4  C. The supernatant,
i.e. the cell free extract was collected and used to synthesize
cadmium sulphide nanoparticles.

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solution. The uorescence spectrum of the sample was measured


on a Jobin Yvon Fluorolog 311 spectrophotometer. The morphology of the nanoparticles was determined by high resolution
scanning electron microscopy (HR-SEM) on a FEI Quanta FEG 200
equipped with energy dispersive X- Ray spectrometer (EDX) at an
accelerating voltage of 20 kV. A small volume of the sample was
drop casted on the carbon tape and allowed to dry under ambient
conditions. EDX is useful in determining the chemical composition
of the nanoparticles. Particle size was determined by high
resolution transmission electron microscopy (HR-TEM) analyses
on a FEI Tecnai G2, model T-30 S-Twin operating at 200 kV. Dilute
solutions of the sample were drop casted onto carbon-coated
copper and allowed to dry at room temperature. The structural
properties of the nanoparticle were obtained by powder X-ray
diffraction. The nanoparticles were dried completely and the
diffractogram was recorded on a Rigaku MiniFlex II diffractometer
with a Cu Ka (1.5406 ) source running at 30 kV and 2u values
ranging from 20 to 80 . Particle size and zeta potential was
measured on a Malvern Zetasizer Nano ZS system. Prior to analyses
the samples were diluted and ltered through a 0.2 mm lter and
analyzed in a range from 0.1 nm to 1000 nm. Fourier transform
infrared (FTIR) spectroscopy analyses was performed using a
BRUKER RFS-27, Stand alone FT Raman Spectrophotometer. Sample
preparation involved lyophilization of the nanoparticles, after
which they were mixed with potassium bromide (KBr) and made
into a tablet with the aid of a bench press. Scanning was performed
between 4000 cm1 to 400 cm1 at a resolution of 2 cm1 in the
transmittance mode.
2.4. Photocatalytic experiments
The degradation of methylene blue in the presence of CdS
semiconductor nanoparticles under UV light was evaluated.
10 ppm of the dye was dissolved in 100 ml of double distilled
water, to which 75 mg of the photocatalyst (CdS nanoparticles) was
added. The solution was vigorously dispersed and kept in the dark
for 30 min to ensure the adsorptiondesorption equilibrium was
reached. This solution was then placed under a UV (Phillips TUV30W) lamp with constant stirring. Samples were periodically taken
and centrifuged to remove the nanoparticles from the dye solution.
 
The concentration of the dye solution CC0 was measured using a
UVvis spectrophotometer in a range from 400800 nm after
separating CdS catalyst from the mixture.
3. Results and discussion

2.2. Synthesis of cadmium sulphide nanoparticles

3.1. Structural, morphological and elemental analyses

25 ml of the cell-free extract was made up to 100 ml with deionized water; this was placed in a water bath set at 65  C.
Cadmium chloride and sodium sulphide were added to the extract
to obtain a nal concentration of 1 mM. The reaction was
completed in twenty minutes, following which the cadmium
sulphide nanoparticles were separated by centrifuging at 8720 g.
The pellet was washed repeatedly (three times) with de-ionized
water to remove other organic matter that may be bound to the
nanoparticles.

Following the addition of the reagents to the cell free extract


there was a progressive change in colour from yellow to orange
indicating the formation of cadmium sulphide nanoparticles. After
the completion of the reaction, the nanoparticles were separated
and washed repeatedly with de-ionized water. These nanoparticles
were used in further experiments. Powder XRD measurements
were performed to determine the structure of the cadmium
sulphide nanoparticles. The observed peaks were indexed with
JCPDS standards and all the reections shown in Fig. 1 can be
indexed with cubic CdS (JCPDS No. 890440). The XRD pattern
exhibited three prominent peaks at 2u values of 26.4 , 44.7 and
51.7 ; these corresponded to the (111), (2 2 0) and (3 11) diffraction
planes of cubic zinc blende phase of cadmium sulphide. From the
diffractogram it is evident that the sample is clearly crystalline and
there are no impurities as there are no unmatched peaks. The
lattice cell parameter was determined to be: a = 5.825 and is in
good agreement with JCPDS card no. 890440. The relatively broad

2.3. Characterization of the CdS nanoparticles


The optical properties of the nanoparticles were studied using
UVvis absorption spectroscopy on a Perkin Elmer (l 35)
spectrophotometer. The absorption was measured from 300
650 nm at a resolution of 1 nm. The samples were diluted with deionized water to avoid errors due to high optical density of the

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M.D. Rao, G. Pennathur / Materials Research Bulletin 85 (2017) 6473

Fig. 1. Powder XRD patterns of cadmium sulphide nanoparticles synthesized using


Chlamydomonas reinhardtii cell free extract.

peaks indicated the ne size of the grains; the particle size was
determined by measuring the fwhm (full width half maximum)
and calculated using the Debye-Scherrer formula.
D

0:9l
bCosu

Where D represents mean crystallite size, l is the wavelength of


the copper target that was used. b represents the full width half
maximum (FWHM) of the peak and is the diffraction angle. The

average grain size from XRD analysis was 6 nm, which was roughly
similar to the sizes measured using HR- TEM. However, the XRD
results overestimated the size of the nanoparticles, this disagreement in the results from two different techniques is justiable
because the XRD line broadening does not take into account other
factors such as lattice defects and strain [10].
The size and morphology of the nanoparticles were analyzed
using HR- TEM, large numbers of spherical nanoparticles were
observed in the HR- TEM micrographs (Fig. 2). Over 50 particles
were measured using ImageJ software and the particles ranged
from 2 to 7 nm. Average particle size was determined to be 4.8 nm.
Possible reasons for the appearance of aggregates could be due to
the large concentration of sample loaded on the grid as well as the
duration of probe sonication applied to disperse the powdered
sample into solution [6]. However, the lattice fringes can be clearly
distinguished (Fig. 2D) indicating the crystalline nature of the
nanoparticles. The lattice fringe spacing for the sample is 3.36 ,
which corresponds to the (1 1 1) phase of cubic zinc blende
structure. The selective area diffraction pattern (SAED) pattern
showed bright concentric rings and is indicative of the crystalline
nature of the nanoparticles (Fig. 2E). Intracellular synthesis of
cadmium sulphide nanoparticles using Escherichia coli demonstrated similar results with narrower size distribution (25 nm)
[30]. Whereas, extracellular synthesis using fungi produced larger
sized particles ranging from 520 nm [23,1]. Interestingly, the algal
cell free extract was able to produce comparatively smaller sized
particles in spite of using extracellular material.
HR-SEM is capable of functioning at high magnications
making it an important tool in determining the topography of
the as-synthesized K1 and is therefore suited to image nanometer
sized objects. The HR-SEM image as seen in Fig. 3 indicated the
formation of large number of spherical nanostructures that were

Fig. 2. HR-TEM images of cadmium sulphide nanoparticles synthesized using Chlamydomonas cell free extract. (AC) HR-TEM micrographs from lower to higher
magnications; (D) Images of CdS nanoparticles showing lattice fringes, (E) SAED image of CdS nanoparticles.

M.D. Rao, G. Pennathur / Materials Research Bulletin 85 (2017) 6473

Fig. 3. HR-SEM and EDX images of cadmium sulphide nanoparticles synthesized


using Chlamydomonas reinhardtii cell free extract.

densely packed. Elemental analysis performed using EDX revealed


the presence of two intense peaks corresponding to cadmium and
sulphur. The other peaks corresponding to oxygen and phosphorus
are most likely due to the organic capping material that is bound to
the surface of the nanoparticles. The peaks at 3.14 keV and 3.37 keV
corresponded to the L1 and L1 peaks of cadmium while the peak at

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around 2.3 keV corresponded to the K1 peak of sulphur [22].


Quantitative analysis of the atomic ratio of cadmium and sulphur
in the sample was nearly stoichiometric (1:1).
The particle size distribution was measured using DLS and the
particles were found to range from 6 to 80 nm (Fig. 4A). The
average particle size corresponding to the maxima was 38 nm. The
hydrodynamic radius that is measured includes the inorganic core
as well as the organic shell and so may not be an accurate reection
of the actual size of the particle. The size of the organic shell
depends on various factors such as solvation layers, excluded
volume interactions, polyelectrolyte effects and restrictions in
bond and rotation angles [13]. Another factor that may have also
inuenced the DLS measurements is the polydispersity index that
was measured at 0.278 indicating that the solution contained
particles of different sizes. Another study also observed variation in
the sizes of cadmium sulphide nanoparticles synthesized using
surfactants when measured using DLS; sizes ranged from 4 to
85 nm depending on the surfactant used [10]. The variation in size
was attributed to the measurement of the adsorbed surfactant
layer/micelles as well as the hydration shell of the head group. Zeta
potential is a physical property of any particle in suspension and
the magnitude of the zeta potential is used as an indicator of the
stability of colloidal system [10]. When nanoparticles in suspension have either a sufciently large positive or negative zeta
potential they would repel each other preventing their aggregation
and this is generally due to interparticle electrostatic repulsion.
However, if the particles have low zeta potential values then there
is no force preventing them from coming together and occulating.
The zeta potential of CdS nanoparticle solution was measured to be
30.7 mV indicating the stability of the solution as observed in
Fig. 4B. The highly negative charge is due to the attachment of
protein functional groups (
NH2 & 
COOH) on the surface of the
nanoparticle. A number of factors inuence the adsorption of
proteins on the nanoparticle surface, these include the size, shape,
composition and surface charge on the surface of the nanoparticle
[24].
3.2. Optical properties of CdS nanoparticles
The optical properties of the synthesized CdS nanoparticles
were monitored using UVvis spectroscopy as seen in Fig. 5A. A
shoulder peak was observed around 430 nm, which is considerably
blue- shifted in comparison to bulk CdS (515 nm) due to quantum
connement. Decrease in particle size increases the energy of
separation between the ground and excited electronic states

Fig. 4. (A) Hydrodynamic size distributions of cadmium sulphide nanoparticles with an average hydrodynamic radius of 38 nm as measured by DLS; (B) Zeta potential of
cadmium sulphide nanoparticles synthesized using the cell free extract of the microalga Chlamydomonas reinhardtii.

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M.D. Rao, G. Pennathur / Materials Research Bulletin 85 (2017) 6473

Fig. 5. (A) UVvis spectrum of cadmium sulphide nanoparticles synthesized using Chlamydomonas reinhardtii cell free extract; (B) Tauc Plot of cadmium sulphide
nanoparticles for band gap determination.

resulting in a blue-shift in absorption. UVvis spectrum showed


broad distribution that suggested an apparent contradiction in
measurements between UVvis spectroscopy, XRD and TEM. This
was likely due to the fact that UVvis spectroscopy has the ability
to register larger particles as well as aggregates. Further,
techniques like TEM measure the actual size of the nanoparticle
and are usually able to distinguish individual particles. A variety of
techniques have been used to measure these nanoparticles and
differences in estimation are not due to measurement error, rather
due to the specicity of each technique.
The optical properties of CdS nanoparticles that had been
synthesized using Fusarium oxysporum biomass reported a peak
around 450 nm, which is red-shifted compared to our results. The
magnitude of the shift depends on the size of the semiconductor,
with smaller particles showing greater shift to shorter wavelengths [16]. The particles synthesized in that study measured
around 20 nm [1]. The average size of CdS nanoparticles was
estimated from the UVvis spectrum using Hengleins empirical
model [33]. Where, particle size i.e. the diameter of the
nanoparticle (2R) is correlated with the exciton absorption
transition onset (lexc) as shown in Eq. (2).
2

Fig. 6. Fluorescence emission spectra of cadmium sulphide nanoparticles synthesized using Chlamydomonas reinhardtii cell free extract.

Based on the UVvis spectra, the average particle size of the CdS
nanoparticles was estimated to be 3.06  0.1 nm indicating that
particle size had reduced by about 45% in comparison to bulk (5.6
3.06 nm) based on quantum connement. This phenomenon could
be due to the stabilizing inuence of algal proteins that act as
capping agents. The optical band gap energies for the assynthesized CdS nanoparticles were evaluated using the Tauc
relation [10]. The average band gap values were determined from
the linear intercept of the Tauc plot on the hn axis as shown in
Fig. 5b and was found to be 2.93  0.02 eV. This value is higher than
the reference bulk value for CdS, which is 2.42 eV, the difference in
band gap energies was measured to be 0.51 eV. This increase in
energy is referred to as a Blue shift and is due to the bound state
of electron-hole pairs in the quantum conned semiconductor
nanoparticle [13]. To the best of our knowledge there are no reports
on the aqueous synthesis of CdS nanoparticles using algal
biomolecules as capping agents.
Fluorescence emission in semiconductor nanoparticles is
generally due to contributions from excitonic recombination,
which usually occurs at the absorption edge and due to emissions

from trapped defects [32]. Fig. 6 shows the photoluminescence


spectrum of the cadmium sulphide nanoparticles excited at
370 nm. The nanoparticles showed two peaks at 410 nm and
430 nm that would correspond to band edge emissions and a
shoulder peak at 470 nm. Yang et al. studied the optical properties
of bovine-haemoglobin conjugated CdS nanocrystals and
uncapped CdS; emission peaks at 510 and 540 nm was observed
[35]. The blue shift in the emission peak was ascribed to the
quantum connement of the exciton as the particle size decreases.
The relative broadness of the emission peak is due to slight
heterogeneity in particle size. The absence of appreciable
emissions at longer wavelengths (500700 nm) due to surface
traps indicated the absence of defect related emission as observed
from the spectrum. This could be attributed to surface modication
by algal biomolecules that stabilize the nanoparticle and improve
photoluminescence intensity. A recent study that functionalized
CdS nanorods with neutravidin observed that optimization of the
ligands that coat the surface of the quantum dots and the
associated conditions play a critical role in improving photoluminescence [11]. More detailed study of the optical properties of

2R

0:1
0:1338  0:0002345  lexc

M.D. Rao, G. Pennathur / Materials Research Bulletin 85 (2017) 6473

these nanoparticles would offer insights regarding its potential as a


biomarker.
The FTIR spectrum of both the algal cell free extract and
cadmium sulphide nanoparticles is shown in Fig. 7A and B. The FTIR spectrum of algal cell free extract showed prominent peaks at
3437 cm1 and 2920 cm1 that indicated the presence of bonds
due to OH stretching and aldehydic CH stretching. Further, a
peak at 1635 cm1 that corresponds to amide I, a result of the
carbonyl stretch in proteins was also observed. FTIR analysis of the
nanoparticle pellet showed a number of peaks at 3409 cm1,
1582 cm1, 1403 cm1, 1155 cm1, 1005 cm1 and 574 cm1 as can
be seen in Fig. 7. The broad peak at 3409 cm1 may be assigned to
the O
H stretch of hydroxyl groups and the N
H stretch of
amines in proteins. The sharp peak at 1584 cm1 corresponds to
N
H bending vibrations, while the peak at 1403 cm1 could be
due to the CH3 and CH2 groups in proteins [18,9]. The increase in
the absorption band corresponding to amide II indicates that the
NH moieties are directly interacting with the nanoparticle surface.
There was an enhancement in absorption around 1400 cm1
indicating the involvement of side chain amino groups [7]. The
band at 1155 cm1 could correspond to C
O
C stretching in
carbohydrates and polysaccharides [5]. The change observed in
nanoparticle spectrum offers evidence of the conjugation of algal
proteins directly on the surface of CdS nanoparticles.
3.3. Mechanism of CdS nanoparticle biosynthesis
Chlamydomonas reinhardtii extract is rich in a number of
proteins, carbohydrates and vitamins. They also contain a number
of oxidoreductases, these enzymes catalyze oxidation and reduction reactions in the cell. It is likely that enzymes that form a part of
the oxidoreductive machinery are responsible for the biotransformation resulting in the formation of CdS nanoparticles. We have
elucidated a plausible mechanism to help understand the
biosynthesis of CdS nanoparticles as depicted in Fig. S1 in
Supplementary material. When CdCl2 is added, Cd2+ ions dissociate
and bind with algal proteins (oxidoreductases) that usually
respond to metallic stress. The subsequent addition of Na2S
results in the formation of S2 ions that bind to the cadmiumbound algal proteins, giving rise to the formation of CdS nuclei. At
higher pHs these enzymes act as reductases and help in the

Fig. 7. FTIR spectrum of (A) algal cell free extract and the (B) as-synthesized
cadmium sulphide nanoparticles with the characteristic peaks.

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synthesis and stabilization of the nanoparticles. The ability to


reduce sulphates to thiols that are eventually assimilated into
amino acids is abundantly present in Plants and microalgae [25].
Chlorella, a unicellular microalgae similar to Chlamydomonas
reinhardtii was found to be rich in thiosulfonate reductase, an
enzyme that catalyzed the reduction of bound sulte to bound
sulphide. They observed that the enzyme transferred the free S2
to Cd2+ resulting in the formation of CdS; ferredoxin was found to
act as an electron donor [26]. A similar mechanism is likely present
in Chlamydomonas reinhardtii, a prototroph that is rich in APS
(5-adenylylsulfate) reductase and sulte reductases [21]. An earlier
report studied the protein prole of the algal cell extract using SDS
PAGE and observed that it contained a number of proteins that
were part of the oxidoreductive machinery; these included super
oxide dismutases as well as oxygen evolver proteins [20].
3.4. Photocatalytic experiments
The photocatalytic activity of biologically synthesized cadmium
sulphide nanoparticles was evaluated by studying the photocatalytic degradation of methylene blue under UV light. Methylene
blue is an organic dye that is frequently used in textile industries- it
is used to dye cotton, silk and wood. Contact with eyes is very
harmful and may lead to loss of vision. Ingestion of the dye leads to
nausea, vomiting, mental disorientation and methemoglobinemia
[19]. The concentration of the active species was determined by
measuring changes in the absoprtion peak at 661 nm over the
course of 90 min. The absorption spectrum of the dye after
different time intervals is shown in Fig. 8. There was an appreciable
decrease in methylene blue concentration within 30 min and
almost complete degradation of the dye at the end the reaction.
The kinetics of the reaction was investigated to get a better
understanding of the photocatalytic degradation process. Photocatalytic oxidation of organic pollutants generally obeys LangmuirHinshelwood kinetics [37]. This kind of pseudo-rst order kinetics
may be represented by the following equation:
 
C
5
K app t
In
C0
Where C is the concentration of the dye, C0 is the initial
concentration of the dye at the start of the reaction; t represents
time and kapp is the rate constant. The rate of the reaction was

Fig. 8. UVvis spectra of the photocatalytic degradation of methylene blue by CdS


nanoparticles as a function of time.

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M.D. Rao, G. Pennathur / Materials Research Bulletin 85 (2017) 6473

 
determined by plotting In CC0 as shown in Fig. 9 and corresponds
to the slope of the tted curve. The rate constant was measured to
be 0.018 min1 and was found to surpass results obtained with a
composite catalyst composed of CdS cross-linked with chitosan
[39]. Our results were comparable with an earlier study that
employed a biotemplated approach for the synthesis of CdSbacterial cellulose derived hybrid nanobres in photocatalysis
under visible light [37]. An earlier report on the degradation of
methylene blue by CdS and ZnS nanoparticles reported a rate
constant of 0.00361 min1 [29]. These results indicate that the
biologically synthesized cadmium sulphide nanoparticles are
better photocatalysts under ultra violet irradiation.
The effect of initial dye concentration on photodegradation was
studied by taking different concentrations of methylene blue
(515 ppm) in the presence of CdS nanoparticles (0.75 g/L) under
UV light. As the concentration of the dye was increased the
photocatalytic efciency correspondingly decreased from approximately 86% to 71% as seen in Fig. 10. The ratio of photocatalytic
degradation can be correlated with the probability of hydroxyl
radicals being generated on the photocatalysts surface to their
reaction with dye molecules [39]. When 5 ppm of dye was used
there was greater degradation of the dye, this could be attributed to
greater interaction between dye molecules and the oxidising
species. As the concentration of the dye was increased, there were
signicantly greater numbers of dye molecules while there is only
a limited amount of hydroxyl radicals, as the catalyst concentration
remains constant. Further, all the active sites on the catalysts
surface would be obscured by the dye, resulting in a loss of
efciency.
Catalyst loading is an important parameter that should be
considered during photodegradation; it inuences the reaction
rate and by extension, the cost of the treatment [4]. The effect of
nano-CdS dosage on the degradation of methylene blue was
investigated in the presence of different amounts of catalyst (37.5
150 mg) with 10 ppm of dye for 90 min of irradiation. The results
are shown in Fig. 11 and as expected an increase in catalyst dosage
was found to augment photodegradation (from 74% to 90%).
Greater catalyst loading meant that there was a higher density of
nanoparticles, which led to improved adsorption of photons and
dye molecules and thereby enhanced degradation [4]. Usually,
when catalyst dosage is increased it may result in a turbid solution,
limiting the penetration of UV light and thus reducing

Fig. 10. Plot of (C/C0) versus time corresponding to the effect of initial dye
concentration in the photodegradation of methylene blue.

Fig. 11. Plot of (C/C0) versus time corresponding to effect of catalytic dosage in the
degradation of methylene blue.

Fig. 9. Plot of ln (C/C0) versus time corresponding to the photocatalytic degradation


of methylene blue.

photoactivation. However, in our study we did not observe a


signicant increase in the opacity of the solution and increasing
catalyst concentration did not hamper light penetration [8].
The photocatalytic performance of the as-synthesized CdS
nanoparticles was evaluated against other cadmium sulphide
nanoparticles under similar experimental conditions [27]. From
Fig. 12 we can discern that the CdS nanoparticles synthesized from
algal extract was far more effective at degrading methylene blue
under the same experimental conditions compared to other
available cadmium sulphide nanoparticles. The as-synthesized
nanoparticles were able to degrade 85% while CdS nanoparticles
that were capped with maltose achieved around 60% efciency and
CdS capped with glucose was only able degrade 50% of the dye.
These results highlight the small size, purity and photocatalytic
superiority of the as-synthesized cadmium sulphide nanoparticles.
This could be ascribed to the proteins that cap the nanoparticle;
these proteins act as an effectual host for dye absorption allowing
for the donor and acceptor molecules to interact. The increased
surface area provided by the proteins allows for more unsaturated

M.D. Rao, G. Pennathur / Materials Research Bulletin 85 (2017) 6473

Fig. 12. Comparison of photocatalytic degradation efciencies of CdS synthesized


from algae, CdS-G and CdS-M. (Reaction conditions: 0.075 g of catalyst, 100 ml of
Methylene Blue dye (10 ppm)).

surface coordination sites that can facilitate absorption of dye


molecules [15]. These experiments showcase their potential as
effective photocatalysts in the degradation of other toxic organic
dyes.
Reusability of a photocatalyst is a vital factor that determines its
stability as well as its potential in practical application. Photocatalytic experiments were carried out using the same catalysts
under identical conditions i.e. 10 ppm of methylene blue dye was
used in the presence of 0.75 g/L of catalyst for a duration of 90 min
under UV light. After the rst round, the solution was centrifuged;
the pellet containing the nanoparticles was washed and reused.
Over the course of ve consecutive cycles there was a marginal
decrease in catalytic efciency from 85.9% after the rst run, to
83.5% following the second cycle, 81.3%, 79.8% and nally 77% after
the fth cycle as observed in Fig. 13. The marginal loss in activity
could be due to some loss of photocatalyst during repeated
centrifugation and washes. However, these results demonstrate
the stability and potent photocatalytic activity of the biologically
synthesized CdS nanoparticles.
An important consideration that should be taken into account
with regard to photocatalytic efciency of CdS nanoparticles is
their photostability. These nanoparticles are usually prone to
photocorrosion due to the rapid recombination of electrons and

71

holes. Charge separation of electron and holes improves photoreactivity and inhibits photocorrosion [28]. To assess the photostability and anti-photocorrosion potential of the biogenically
synthesized CdS nanoparticles the concentration of elemental
sulphur as well as Cd2+ released into the solution after the
degradation of methylene blue was measured. After ve rounds of
recycling the photocatalyst, elemental sulphur concentration had
marginally increased from 1.5 ppm to little more than 3.5 ppm
(Fig. 14A). While Cd2+ concentration slowly increased from
approximately 2 ppm to 9.5 ppm (Fig. 14B) by the end of the fth
run. An earlier report that compared the photostability of bare CdS
as well as a composite composed of CdS nanoparticles intercalated
between titanate sheets found that bare CdS released 62.5 ppm
while the composite released around 15 ppm [28]. These results
demonstrate the inherent photostability of the algal-synthesized
nanoparticles. This could be attributed to the proteins capping the
nanoparticles, which enhance charge transfer from the nanoparticle to the attached protein thereby reducing recombination of
electrons and holes and thus improving photocatalytic ability of
these nanoparticles.
Active species trapping experiments were conducted to gain
insights into the photocatalytic mechanism of biosynthesized CdS
nanoparticles under UV light. 2-propanol (0.2 mol/L) was added
the dye solution to act as a hydroxyl radical (OH) scavenger, while
benzoquinone (BQ, 0.02 mol/L) was added to capture superoxide
radicals (O2) respectively. Disodium ethylenediaminetetraacetate (EDTA-2Na; 0.05 mol/L) was added to scavenge photogenerated holes [40]. Photocatalytic activity of the nanoparticles was
signicantly inhibited following the addition of EDTA-2Na,
indicating that photogenerated holes play a key role in the process
as seen in Fig. 15. Addition of EDTA degraded approximately 38% of
the dye while in its absence the photodegradation efciency
increased to 86% as seen in Fig. 15. 2-Propanol was also found to
inhibit the nanocatalysts ability to breakdown methylene blue by
reducing efciency to approximately 55% while benzoquinone was
found to play a less signicant role in dye degradation. These
results indicate that OH radicals as well as photogenerated holes
play an important role in the UV-light driven photocatalytic
degradation of methylene blue.
Irradiation of CdS nanoparticles produces a number of electronhole pairs that are powerful oxidising and reducing agents. These
holes are able to oxidize adsorbed water and produce hydroxyl
radicals.
CdS + hn ! h++ e

(3)

h+ + H2O ! OH + H+

(4)

Furthermore, oxygen helps prevent the recombination of


electron-hole pairs [27]. These hydroxyl radicals act as oxidizing
agents mineralizing methylene blue to harmless byproducts that
include CO2 and H2O. Further, electrons in the conduction band are
taken up by oxygen thus producing anionic superoxide radicals
that are involved in further oxidation.
MB + OH/O2 ! Harmless products (CO2 + H2O + NH4+ + NO3 +
SO42 + Cl)
(5)
4. Conclusion

Fig. 13. Plot depicting the reusability of CdS photocatalysts in the photodegradation
of methylene blue.

This work discusses the green synthesis of CdS nanoparticles


from the aqueous cell free extract of Chlamydomonas reinhardtii.
Spherical particles approximately measuring 5 nm were observed
under HRTEM. FTIR experiments indicated that algal biomolecules
are involved in the synthesis and stabilization of these

72

M.D. Rao, G. Pennathur / Materials Research Bulletin 85 (2017) 6473

Fig. 14. Estimation of (A) Elemental sulphur and (B) Photocorrosion of CdS after ve consecutive cycles of photodegradation of methylene blue.

References

Fig. 15. Effect of scavengers on the photocatalytic degradation of methylene blue


under visible light radiation.

nanoparticles. Optical analyses of the nanostructures showed an


increase in the band gap to 2.93 eV. Photoluminescence spectrum
of CdS nanoparticles revealed emission peaks at 430 nm and
470 nm. The nanoparticles were found to be stable in solution with
a zeta potential of 30.7 mV. The synthesized semiconductor
nanoparticles were found to be efcient photocatalysts of organic
dyes. Their reusability and photostability are important aspects for
practical consideration and these nanoparticles showed excellent
photostability. This method has potential for extensive synthesis of
CdS nanoparticles by an environmentally benign approach and its
application in the photodegradation of organic dyes.
Acknowledgements
The authors are very grateful for the support extended by the
Department of Science and Technology and the Department of
Biotechnology. MDR would like to thank SAIF, IIT Chennai for help
with HRSEM and FTIR experiments. MDR is very thankful to D.
Sudha for her continued support.
Appendix A. Supplementary data
Supplementary data associated with this article can
be found, in the online version, at http://dx.doi.org/10.1016/j.
materresbull.2016.08.049.

[1] A. Ahmad, P. Mukherjee, D. Mandal, S. Senapati, M.I. Khan, R. Kumar, M. Sastry,


Enzyme mediated extracellular synthesis of cds nanoparticles by the fungus,
fusarium oxysporum, J. Am. Chem. Soc. 124 (2002) 1210812109.
[2] L. Castro, M.L. Bla zquez, J.A. Mun oz, F. Gonz alez, A. Ballester, Biological
synthesis of metallic nanoparticles using algae, IET Nanobiotechnol. 7 (2013)
109116.
[3] M.N. Chong, B. Jin, C.W. Chow, C. Saint, Recent developments in photocatalytic
water treatment technology: a review, Water Res. 44 (2010) 29973027.
[4] M. Darwish, A. Mohammadi, N. Assi, Microwave-assisted polyol synthesis and
characterization of pvp-capped cds nanoparticles for the photocatalytic
degradation of tartrazine, Mater. Res. Bull. 74 (2016) 387396.
[5] J. Jena, N. Pradhan, R.R. Nayak, B.P. Dash, L.B. Sukla, P.K. Panda, B.K. Mishra,
Microalga scenedesmus sp.: a potential low-cost green machine for silver
nanoparticle synthesis, J. Microbiol. Biotechnol. 24 (2014) 522533.
[6] J. Jiang, G. Oberd orster, P. Biswas, Characterization of size, surface charge, and
agglomeration state of nanoparticle dispersions for toxicological studies, J.
Nanopart. Res. 11 (2009) 7789.
[7] A.A. Kamnev, Infrared spectroscopy in studying biofunctionalised gold
nanoparticles, Nanomaterials Imaging Techniques, Surface Studies, and
Applications, Springer, 2013, pp. 3550.
[8] S. Kansal, M. Singh, D. Sud, Studies on photodegradation of two commercial
dyes in aqueous phase using different photocatalysts, J. Hazard. Mater. 141
(2007) 581590.
[9] M.K. Kim, K.H. Jeune, Use of ft-ir to identify enhanced biomass production and
biochemical pool shifts in the marine microalgae, chlorella ovalis, cultured in
media composed of different ratios of deep seawater and fermented animal
wastewater, J. Microbiol. Biotechnol. 19 (2009) 0107.
[10] S. Kumar, M. Gradzielski, S. Mehta, The critical role of surfactants towards cds
nanoparticles: synthesis, stability: optical and pl emission properties, RSC Adv.
3 (2013) 26622676.
[11] L.G. Lippert, J.T. Hallock, T. Dadosh, B.T. Diroll, C.B. Murray, Y.E. Goldman,
Neutravidin functionalization of cdse/cds quantum nanorods and
quantication of biotin binding sites using biotin-4- uorescein uorescence
quenching, Bioconjugate Chem. 11 (3) (2016) 562568.
[12] T. Mamedov, V. Yusibov, Green algae chlamydomonas reinhardtii possess
endogenous sialylated n-glycans, FEBS Open Biol. 1 (2011) 1522.
[13] A.A. Mansur, S.M. de Carvalho, H.S. Mansur, Bioengineered quantum dot/
chitosan-tripeptide nanoconjugates for targeting the receptors of cancer cells,
Int. J. Biol. Macromol. 82 (2016) 780789.
[14] J. Mao, X.M. Chen, X.W. Du, Facile synthesis of three dimensional cds
nanoowers with high photocatalytic performance, J. Alloys Compd. 656
(2016) 972977.
[15] N. Jain, A. Bhargava, J. Panwar, Enhanced photocatalytic degradation of
methylene blue using biologically synthesized protein-capped ZnO
nanoparticles, Chem. Eng. J. 243 (2014) 549555.
[16] M. Pal, N. Mathews, P. Santiago, X. Mathew, A facile one-pot synthesis of highly
luminescent cds nanoparticles using thioglycerol as capping agent, J.
Nanopart. Res. 14 (2012) 113.
[17] V. Pardo-Yissar, E. Katz, J. Wasserman, I. Willner, Acety- choline esteraselabeled cds nanoparticles on electrodes: photoelectron-chemical sensing of
the enzyme inhibitors, J. Am. Chem. Soc. 125 (2003) 622623.
[18] D. Pathania, S. Sarita, B.S. Rathore, Synthesis: characterization and
photocatalytic application of bovine serum albumin capped cadmium
sulphide nanoparticles, Chalcogenide Lett. 8 (2011) 396404.
[19] M. Rafatullah, O. Sulaiman, R. Hashim, A. Ahmad, Adsortion of methylene blue
on low-cost adsorbents: a review, J. Hazard. Mater. 177 (2010) 7080.
[20] D. Rao, P. Gautam, A facile one-pot synthesis of gold nanoparticles by
Chlamydomonas reinhardtii, Asian J. Microbiol. Biotechnol. Environ. Sci. 16
(2015) 633639.
[21] C.G. Ravina, C.I. Chang, G.P. Tsakraklides, J.P. McDermott, J.M. Vega, T. Leustek,
C. Gotor, J.P. Davies, The sac mutants of chlamydomonas reinhardtii reveal

M.D. Rao, G. Pennathur / Materials Research Bulletin 85 (2017) 6473

[22]

[23]
[24]

[25]
[26]
[27]

[28]

[29]

[30]

[31]

transcriptional and posttranscriptional control of cysteine biosynthesis, Plant


Physiol. 130 (2002) 20762084.
S. Rengaraj, S. Venkataraj, S.H. Jee, Y. Kim, C.W. Tai, E. Repo, A. Koistinen, A.
Ferancova, M. Sillanpa, Cauliower-like cds microspheres composed of
nanocrystals and their physicochemical properties, Langmuir 27 (2010)
352358.
R. Sanghi, P. Verma, A facile green extracellular biosynthesis of cds
nanoparticles by immobilized fungus, Chem. Eng. J. 155 (2009) 886891.
A. Sankhla, R. Sharma, R.S. Yadav, D. Kashyap, S. Kothari, S. Kachhwaha,
Biosynthesis and characterization of cadmium sulde nanoparticlesan
emphasis of zeta potential behavior due to capping, Mater. Chem. Phys. 170
(2016) 4451.
J.A. Schiff, R.C. Hodson, Pathways of sulfate reduction in algae, Ann. N.Y. Acad.
Sci. 175 (1970) 555576.
A. Schmidt, Sulfate reduction in a cell-free system of chlorella, Arch. Microbiol.
93 (1973) 2952.
M. Shakir, M. Faraz, M.S. Khan, S.I. Al-Resayes, The photocatalytic: in vitro
anthelmintic activity of biomolecule-inspired CDS nanoparticles, C.R. Chim. 9
(2015) 966978.
S. Sehati, M.H. Entezari, Sono-intercalation of CdS nanoparticles into the layers
of titanate facilitates the sunlight degradation of Congo red, J. Colloid Interface
Sci. 462 (2016) 130139.
N. Soltani, E. Saion, M.Z. Hussein, M. Erfani, A. Abedini, G. Bahmanrokh, M.
Navasery, P. Vaziri, Visible light-induced degradation of methylene blue in the
presence of photocatalytic zns and cds nanoparticles, Int. J. Mol. Sci. 13 (2012)
1224212258.
R.Y. Sweeney, C. Mao, X. Gao, J.L. Burt, A.M. Belcher, G. Georgiou, B.L. Iverson,
Bacterial biosynthesis of cadmium sulde nanocrystals, Chem. Biol. 11 (2004)
15531559.
K.N. Thakkar, S.S. Mhatre, R.Y. Parikh, Biological synthesis of metallic
nanoparticles, Nanomed. Nanotechnol. Biol. Med. 6 (2010) 257262.

73

[32] Y. Wang, G. Meng, L. Zhang, C. Liang, J. Zhang, Catalytic growth of large-scale


single-crystal cds nanowires by physical evaporation and their
photoluminescence, Chem. Mater. 14 (2002) 17731777.
[33] H. Weller, H. Schmidt, U. Koch, A. Fojtik, S. Baral, A. Henglein, W. Kunath, K.
Weiss, E. Dieman, Photochemistry of colloidal semiconductors: onset of light
absorption as a function of size of extremely small cds particles, Chem. Phys.
Lett. 124 (1986) 557560.
[34] T. Xaba, M. Moloto, N. Moloto, The effect of water-soluble capping molecules in
the green synthesis of cds nanoparticles using the (z)-2-(pyrrolidin-2-ylidene)
thiourea ligand, Mater. Lett. 146 (2015) 9195.
[35] G. Yang, D. Qin, X. Du, L. Zhang, G. Zhao, Q. Zhang, J. Wu, Aqueous synthesis and
characterization of bovine hemoglobin- conjugated cadmium sulde
nanocrystals, J. Alloys Compd. 604 (2014) 181187.
[36] H. Yang, C. Huang, X. Li, R. Shi, K. Zhang, Luminescent and photocatalytic
properties of cadmium sulde nanoparticles synthesized via microwave
irradiation, Mater. Chem. Phys. 90 (2005) 155158.
[37] J. Yang, J. Yu, J. Fan, D. Sun, W. Tang, X. Yang, Biotemplated preparation of cds
nanoparticles/bacterial cellulose hybrid nanobers for photocatalysis
application, J. Hazard. Mater. 189 (2011) 377383.
[38] J. Zhao, J.A. Bardecker, A.M. Munro, M.S. Liu, Y. Niu, I.K. Ding, J. Luo, B. Chen, A.K.
Y. Jen, D.S. Ginger, Efcient cdse/cds quantum dot light-emitting diodes using a
thermally polymerized hole transport layer, Nano Lett. 6 (2006) 463467.
[39] H. Zhu, R. Jiang, L. Xiao, Y. Chang, Y. Guan, X. Li, G. Zeng, Photocatalytic
decolorization and degradation of congo red on innovative crosslinked
chitosan/nano-cds composite catalyst under visible light irradiation, J. Hazard.
Mater. 169 (2009) 933940.
[40] Y. Zhu, Z. Chen, T. Gao, Q. Huang, F. Niu, L. Qin, P. Tang, Y. Huang, Z. Sha, Y. Wang,
Construction of hybrid Z-scheme Pt/CdSTNTAs with enhanced visible-light
photocatalytic performance, Appl. Catal. B 163 (2015) 1622.