International Journal of Biological Macromolecules 48 (2011) 106–111

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International Journal of Biological Macromolecules

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

Microwave assisted synthesis of polyacrylamide grafted starch (St-g-PAM) and

its applicability as flocculant for water treatment
Sumit Mishra, Ankita Mukul, Gautam Sen ∗ , Usha Jha
Department of Applied Chemistry, Birla Institute of Technology, Mesra, Ranchi 835 215, Jharkhand, India

a r t i c l e i n f o a b s t r a c t

Article history: Polyacrylamide grafted starch (St-g-PAM) was made by a novel method of synthesis, involving combi-
Received 15 September 2010 nation of microwave radiation and a chemical free radical initiator (ceric ammonium nitrate) to initiate
Received in revised form 4 October 2010 grafting reaction. This method (microwave assisted synthesis) is quick, highly reliable, reproducible and
Accepted 7 October 2010
yields high quality product as compared to the conventional method (which uses a chemical free radical
Available online 15 October 2010
initiator alone to initiate the grafting reaction).The St-g-PAM grades synthesized were characterized by
various physicochemical techniques. Further, its application as flocculant for wastewater treatment was
Keywords:
investigated.
Ceric ammonium nitrate
Flocculant © 2010 Elsevier B.V. All rights reserved.
Microwave assisted synthesis
Jar test
Polyacrylamide grafted starch

1. Introduction The best method of graft copolymer synthesis is by use of

The importance of graft copolymers in the world of polymer sci-
ence is analogous to the importance of alloying in case of metals. In
case of grafted polysaccharides this assumes even greater impor-
tance as grafting is perhaps the most effective way of regulating the
properties of polysaccharides ‘tailor-made’ according to our needs.
The main problem in case of graft copolymers is the lack of com-
mercial methods of synthesis. The chief methods of synthesis of
grafted polysaccharides involves use of chemical free radical ini-
tiator (conventional method), high energy radiations (gamma and
X-ray), UV-radiation based method and microwave based methods.
The conventional method of synthesis uses a chemical free rad-
ical initiator (e.g. ceric ammonium nitrate or CAN) to generate free
radical sites on the backbone polymer, where the monomer of the
graft gets added up to form the graft chain [1–5]. This method
of synthesis has low reproducibility and is not very suitable for
commercial scale synthesis.
A better method of graft copolymer synthesis is by use of high
energy radiation (gamma rays or electron beam) as the free radical
generator [6–7]. But this method is not suitable for grafted polysac-
charide synthesis as the high energy radiation can cause damage to
the polysaccharide backbone (radiolysis).
UV rays in presence of suitable photosensitizor can also be used,
but low penetration of UV-rays makes it suitable for surface grafting
only.
∗ Corresponding author. Tel.: +91 9470137364.
E-mail address: gsen9@hotmail.com (G. Sen).
microwave radiation to generate the free radical sites on the back-
bone polymer. Recently, the microwave based methods of graft
copolymer synthesis has been classified into two types [8]:
(1) Microwave initiated synthesis: uses microwave radiation alone
to create free radical sites on the polysaccharide backbone, from
where the graft chains grow [4,8–10].
(2) Microwave assisted synthesis: uses a combination of
microwave radiation and chemical free radical initiator to
create the free radical sites on the polysaccharide backbone,
from where the graft chains grow.
In the study reported here, we have synthesized polyacrylamide
grafted starch (St-g-PAM) by ‘microwave assisted’ method, in pres-
ence of ceric ammonium nitrate (CAN) as free radical initiator.
Microwave assisted synthesis of grafted starch using CAN as free
radical initiator, has never been reported before to the best of our
knowledge.
Grafted polysaccharides have diverse applications, cutting
across various fields of science and technology. Some important
applications are as flocculant for wastewater treatment [4,9], as
matrix for controlled drug release [8,10] and rheological appli-
cations. In this study, we have investigated the flocculation
efficacy of polyacrylamide grafted starch (St-g-PAM) synthesized
by microwave assisted process, towards its application in wastew-
ater treatment.
Water is the single most important resource that decides the
fate of human being and for that matter any living being. The
present ever increasing population and improving standard of liv-

0141-8130/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.ijbiomac.2010.10.004
 S. Mishra et al. / International Journal of Biological Macromolecules 48 (2011) 106–111
ing, coupled with depleting water resources have put a strong
case on recycling of wastewater. Flocculation is an effective way
of treating wastewater as it not only removes the colloidal parti-
cles (as floc) but also the heavy metals and other contaminants that
gets adsorbed in them. Further, these flocculated colloidal parti-
cles include most of the microbes present in the wastewater. Also,
removal of colloidal particles makes sterilization of the water easy
as the microbes cannot get shelter against the colloidal particles
and hence gets full exposure to the sterilizing radiations.
Flocculation is a technique where polymers are involved in a
solid–liquid separation by an aggregation process of colloidal par-
ticles [11]. Both synthetic and natural polymers have been utilized
for flocculation. The advantage of flocculation over coagulation is
that the former uses only polymeric materials in minute quanti-
ties. These polymeric materials are mostly physiologically inert and
are biodegradable. On the other hand, coagulation involves use of
inorganic salts (e.g. alums) for charge neutralization of colloidal
particles. These inorganic salts are required to be added in higher
doses and their very presence in the water can lead to detrimental
effects (e.g. aluminium exposure due to use of alum as coagulant,
can lead to Alzheimer’s disease over long term [12]).
2. Materials and methods:
2.1. Materials
Maize starch and ceric ammonium nitrate were supplied by E.
Merck (India) Limited, Mumbai, India. Acrylamide was procured
from E. Merck, Germany. Analar grade of acetone and hydroquinone
was purchased from S.D. Fine Chemicals, Mumbai, India. All the
chemicals were used as received; without further purification.
2.2. Synthesis
2.2.1. Microwave assisted synthesis of polyacrylamide grafted
starch (St-g-PAM), using ceric ammonium nitrate (CAN) as free
radical initiator
1 gm of starch was dissolved in 40 ml distilled water. Desired
amount of acrylamide was dissolved in 10 ml water and was added
to the starch solution. They were mixed well and were transferred
to the reaction vessel (1000 ml borosil beaker) and catalytic amount
of ceric ammonium nitrate (CAN) was added. The reaction vessel
was subsequently placed on the turntable of a microwave oven.
Now, microwave irradiation at 800 W of power was performed.
Periodically, the microwave irradiation was paused just before
boiling of the reaction mixture starts (∼65 ◦ C) and was cooled by
placing the reaction vessel in cold water. This was done to keep
the competing reaction of homopolymer formation to the mini-
mum and also to prevent formation of vapours, which may contain
acrylamide and hence can be toxic/carcinogenic.
This microwave irradiation – cooling cycle was repeated until
a gel like mass was left or up to 3 min of irradiation time (if no
gelling took place). Once this microwave irradiation process was
complete, the reaction vessel and its contents were finally cooled
and kept undisturbed for 24 h, to complete the grafting reactions.
Now, the gel like mass left in the reaction vessel was poured into
excess of acetone. The resulting precipitate of graft copolymer was
collected and was dried in hot air oven. Subsequently, it was pulver-
ized and sieved. The percentage grafting of this microwave assisted
synthesized St-g-PAM was evaluated as:
wt. of graft copolymer − wt. of polysaccharide
% grafting = 100
wt. of polysaccharide
The proposed mechanism of synthesis has been depicted in
Scheme 1 and the synthesis details of various grades of the graft
copolymer have been shown in Table 1.
107
2.2.2. Purification of the graft copolymer by solvent extraction
method
Any occluded polyacrylamide (PAM) formed by compet-
ing homopolymer formation reaction was removed from the
grafted polymers synthesized as above, by solvent extraction
using a mixture of formamide and acetic acid (1:1 by volume)
[13].
2.3. Characterization
2.3.1. Intrinsic viscosity measurement
Viscosity measurements of the polymer solutions were car-
ried out with an Ubbelodhe viscometer (constant: 0.003899) at
25 ◦ C. The viscosities were measured in aqueous solutions. The
pH of the solution was neutral. The time of flow for solutions
was measured at four different concentrations. From the time
of flow of polymer solutions (t) and that of the solvent (t0 , for
distilled water), relative viscosity (rel = t/t0 ) was obtained. Spe-
cific viscosity was calculated from the relation sp = rel − 1. Then,
the reduced viscosity (sp /C) and the inherent viscosity (ln rel /C)
were calculated, where ‘C’ is the polymer concentration in g/dL.
The intrinsic viscosity was obtained from the point of intersection
after extrapolation of two plots, i.e. sp /C versus C and ln rel /C
versus C, to zero concentration [14]. The intrinsic viscosity of
the various grades of the graft copolymers has been given in
Table 1.
2.3.2. Elemental analysis
The elemental analysis of starch and that of St-g-PAM 2 (best
grade of the grafted starch synthesized) was undertaken with an
Elemental Analyzer (Make – M/s Elementar, Germany; Model –
Vario EL III). The estimation of five elements, i.e. carbon, hydrogen,
nitrogen, oxygen and sulphur were undertaken. The results have
been summarized in Table 2.
2.3.3. FTIR spectroscopy
The FTIR spectrums of starch (Fig. 1(a)) and of St-g-PAM 2
(Fig. 1(b)) were recorded in solid state, by KBr pallet method using
a FTIR spectrophotometer (Model IR-Prestige 21, Shimadzu Corpo-
ration, Japan) between 400 and 4000 cm−1 .
2.3.4. Scanning electron microscopy
Surface morphology of starch (Fig. 2(a)) and St-g-PAM 2
(Fig. 2(b)) were analyzed in scanning electron microscopy (SEM)
in powdered form (Model: JSM-6390LV, Jeol, Japan).
2.3.5. TGA studies
The thermo gravimetric analysis (TGA) of starch and that of the
St-g-PAM 2 were carried out with TGA instrument (Model: DTG-60;
Shimadzu, Japan). The study was performed in an inert atmosphere
(nitrogen) from 25 ◦ C to 600 ◦ C. The heating rate was uniform in all
cases at 5 ◦ C/min. The concerned TGA curves have been shown in
Fig. 3(a) and (b).
3. Flocculation studies
Flocculation efficacies of various synthesized grades of St-g-
PAM were studied by standard jar test procedure, in 0.25% kaolin
suspension.
All flocculation experiments were carried out in jar test appa-
ratus (Make: Simeco, Kolkata, India). The test protocol involved
taking a measured quantity (800 ml) of the 0.25% suspension
in 1000 ml borosil beaker. Calculated amount of the flocculant
(starch or various grades of St-g-PAM) was added in concen-
trated solution form (except in case of blank, where no flocculant
was added) to achieve the desired concentrations (ranging from
108 S. Mishra et al. / International Journal of Biological Macromolecules 48 (2011) 106–111

Scheme 1. Schematic representation for ‘microwave assisted’ synthesis of St-g-PAM.

Table 1
Polyacrylamide grafted starch (St-g-PAM) grades synthesized.

Grade Wt of starch (gm) Wt of acrylamide (gm) Wt of CAN (gm) Time of irradiation (up to % Graftinga Intrinsic viscosity (dl/g)
gel formation) (s)

St-g-PAM 1 1 5 0.3 167 480 1.3

St-g-PAM 2 1 10 0.3 30 907 3.69
St-g-PAM 3 1 15 0.3 30 904 3.11
St-g-PAM 4 1 10 0.2 26 839 2.72
St-g-PAM 5 1 10 0.4 98 900 3.62
Starch (St) – – – – – 0.91
wt. of graft copolymer − wt. of polysaccharide
a
% grafting = wt. of polysaccharide
100

0 ppm to 1 ppm). The solutions were stirred identically (i.e. in turbidity meter (Digital Nephelo-Turbidity Meter 132, Systronics,
‘jar test’ apparatus), at 150 r.p.m for 30 s, 60 r.p.m for 5 min fol- India). The flocculation efficacy thus studied for starch and var-
lowed by 15 min of settling time. Afterwards, supernatant liquid ious grades of St-g-PAM have been graphically compared in
was collected and turbidity measured in a calibrated nephelo- Fig. 4.

Table 2
Elemental analysis results.

Polymer grade %C %H %N %O %S

Starch 38.86 7.36 0.00 53.78 0.00

PAM 50.80 7.69 19.76 21.7 0.00
St-g-PAM 2 42.52 7.99 14.79 34.70 0.00
 S. Mishra et al. / International Journal of Biological Macromolecules 48 (2011) 106–111
Fig. 1. FTIR spectrum of (a) starch and (b) St-g-PAM 2.
4. Results and discussion
4.1. Synthesis of St-g-PAM by microwave assisted method
St-g-PAM has been synthesized by microwave assisted method
(i.e. synthesis based on free radical mechanism using microwave
radiation in synergism with ceric ammonium nitrate, to gener-
ate free radicals on the starch backbone). Various grades of the
graft copolymer were synthesized by varying the ceric ammonium
nitrate (CAN) and acrylamide (monomer) concentration. In each
case, the microwave irradiation of the reaction mixture was con-
tinued until it sets into a viscous gel like mass. The synthesis details
have been tabulated in Table 1. The optimized grade has been deter-
mined through its higher percentage grafting and intrinsic viscosity
(which is proportional to molecular weight). From Table 1, it is
obvious that the grafting is optimized at acrylamide concentration
of 10 gm and CAN concentration of 0.3 gm in the reaction mixture
(∼60 ml), when the microwave power is maintained at 800 W.
The mechanism of microwave assisted grafting has been
depicted in Scheme 1.
The mechanism by which Ce (IV) generates free radical is
believed to involve the formation of a chelate complex between
the hydroxyl group of the polysaccharide and the oxidant. The
complex so formed disproportionates forming free radicals on the
polysaccharide backbone.
The microwave radiation rotates the acrylamide molecules,
leading to elongation of its bonds. As the C–C double bond elon-
109
Fig. 2. SEM micrograph of (a) starch and (b) St-g-PAM 2.
gates, the pi bond electron cloud splits up into two localized clouds
(i.e. free radical sites on the constituent carbon atoms).
The free radical sites thus created on the starch backbone (by
CAN) and on the acrylamide (by microwave radiation) interacts
through usual free radical reaction mechanism, to yield the graft
copolymer.
A series of five grades of the graft copolymers have been syn-
thesized by this method.
Interpretation for using hydroquinone as inhibitor: Inhibitors
such as hydroquinone (HQ) react with chain radicals to terminate
chain propagation and the resulting hydroquinone radical is sta-
ble and cannot initiate further polymerization. The stability of the
hydroquinone radical is because of the delocalization of electron
charge density throughout the aromatic structure. Thus, addition of
hydroquinone quenches the grafting reaction. This inhibitor action
of hydroquinone supports the proposed free radical mechanism
[4,10].
StO∗ + QH → StOH + Q∗
4.2. Characterization
4.2.1. Estimation and interpretation of intrinsic viscosity
The intrinsic viscosity was evaluated for starch and the various
grades of St-g-PAM, as stated in Table 1.
It is obvious that the intrinsic viscosities of all the grades of
St-g-PAM are greater than that of starch. This can be explained
by the higher molecular weight of St-g-PAM than starch, due to
the grafting of the PAM branches on the main polymer backbone.
110 S. Mishra et al. / International Journal of Biological Macromolecules 48 (2011) 106–111
Fig. 3. TGA of (a) starch and (b) St-g-PAM 2.
This is in good agreement with Mark–Houwink–Sakurada relation-
ship (Intrinsic viscosity  = KM˛ , where K and ˛ are constants, both
related to stiffness of the polymer chains).
4.2.2. Elemental analysis
The results of elemental analysis for starch and of the best grade
of polyacrylamide grafted starch (i.e. St-g-PAM 2) are given in
Table 2. The presence of nitrogen in case of St-g-PAM 2 (but not
in starch) confirms that the PAM chains have indeed been grafted
on the backbone of starch.
4.2.3. FTIR spectroscopy
From the FTIR spectrum of starch (Fig. 1(a)), it is being observed
that a broad peak at 3603.03 cm−1 is due to the stretching vibra-
tions of O–H, a smaller peak at 2887.44 cm−1 attributed to the C–H
stretching vibrations. The bands at 985.62 cm−1 and 929.69 cm−1
are assigned to C–O–C stretching vibrations.
In case of St-g-PAM 2 (Fig. 1(b)), O–H stretching band of
hydroxyl group of starch and N–H stretching band of amide
group of PAM overlap with each other and lead to a peak at
3728.40 cm−1 and a shouldered peak at 3628 cm−1 . A small peak at
3030 cm−1 is attributed to the C–H stretching vibration. The bands
at 881.47 cm−1 and 829.39 cm−1 are assigned to C–O–C stretching
vibrations.
The appearance of a sharp peaks at 1791.87 cm−1 and at
1504.48 cm−1 for St-g-PAM 2 are attributed to C O and N–H
Fig. 4. Flocculation characteristics of starch and synthesized grades of St-g- PAM.
stretching (i.e. Amide I and Amide II stretching) respectively. Fur-
ther, there is one more additional peak present in grafted product
i.e. at 1334.74 cm−1 , which is assigned to the C–N stretching. Thus,
the presence of these additional peaks in case of grafted starch
compared to that of starch confirms the successful grafting of PAM
chains onto the backbone of starch.
4.2.4. Scanning electron microscopy (SEM) analysis
It is evident from the SEM micrographs of starch (Fig. 2(a)) and
that of the best grades of St-g-PAM (Fig. 2(b)) that profound mor-
phological change, in form of transition from granular to lace like
structure, have taken place because of grafting of PAM chains onto
starch. Thus, it is evident that the granular morphology of starch is
lost after grafting and transformed into lacy morphology.
4.2.5. Thermal analysis
The TGA curves of starch (Fig. 3(a)) essentially involved two dis-
tinct zones of weight loss. The initial weight loss is at 30 ◦ C–120 ◦ C.
This is due to the traces of moisture present. The second zone
(250 ◦ C–330 ◦ C) was due to degradation of the starch backbone.
St-g-PAM 2 (Fig. 3(b)), in addition to the above zones of weight
loss, have two extra zones of weight loss (340 ◦ C–460 ◦ C and
460 ◦ C–600 ◦ C), due to the grafted PAM chains.
5. Flocculation studies
The flocculation study in 0.25% kaolin suspension in ‘jar test’
apparatus has been graphically represented in Fig. 4.
All the grades of grafted starch have shown better flocculation
efficacy than starch, due to their higher hydrodynamic volume (i.e.
intrinsic viscosity) as reported in Table 1. The higher hydrodynamic
volume of the macromolecule leads to its higher flocculation effi-
cacy (ref: Singh’s easy approachability model [15,16]).
Further, among the various grades of St-g-PAM, the optimized
grade (St-g-PAM 2) showed maximum flocculation efficacy due to
its highest hydrodynamic volume.
 S. Mishra et al. / International Journal of Biological Macromolecules 48 (2011) 106–111
For all the polymers, there is an optimal dosage at which the
flocculation caused by it is maximum (i.e. the turbidity of the super-
natant collected is minimum). Beyond this dosage, the flocculation
decreases (i.e. turbidity of the collected supernatant increases) due
to destabilization of the flocs formed by the excess polymers (floc-
culant). This behaviour of the flocculation curve finely confirms the
bridging mechanism [17,18] involved behind the phenomenon.
According to bridging mechanism, when long chain polymers in
small dosage are added to a colloidal suspension, they get adsorbed
onto two or more particle surfaces and thus form a bridge between
them. There should be sufficient unoccupied space on the parti-
cle surface so as to form the polymer bridging. This phenomenon
is observed upto a particular dosage of polymer beyond which
flocculation diminishes, the process being known as steric stabi-
lization. Thus, at lower dosages of polymer, no significant bridging
occurs between and hence flocculation remains low. Similarly at
higher dosages of polymer, there is insufficient particle surface for
attachment of the polymer segments leading to destabilization of
flocs.
The optimal dosage of St-g-PAM 2 as flocculant, in 0.25% kaolin
suspension is at 0.75 ppm.
6. Conclusion
Polyacrylamide grafted starch (St-g-PAM) has been synthesised
by a novel ‘microwave assisted’ method, in which a combination of
microwave radiations and chemical free radical initiator i.e. ceric
ammonium nitrate (CAN) has been used to synthesize the graft
copolymer. The synthesized grades of the graft copolymer were
111
characterized through various physicochemical techniques. Fur-
ther, the flocculation efficacy of the graft copolymer was studied
through standard ‘Jar test’ procedure in 0.25% kaolin suspension.
It was found that the St-g-PAM grade with highest hydrodynamic
volume (i.e. intrinsic viscosity) showed the maximum flocculation
efficacy, as predicted by ‘Singh’s easy approachability model’.
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