International Journal of Biological Macromolecules 119 (2018) 954–961

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International Journal of Biological Macromolecules

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

Amphiphilic graft copolymeric micelle using dextrin and poly (N-vinyl

caprolactam) via RAFT polymerization: Development and application
Aniruddha Pal, Abanindra Nath Sarkar, Puja Das Karmakar, Sagar Pal ⁎
Department of Applied Chemistry, Indian Institute of Technology (ISM), Dhanbad 826004, India

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

Article history: Dextrin and poly (N-vinyl caprolactam) based amphiphilic graft copolymer has recently been developed using
Received 13 November 2017 RAFT polymerization. The prepared graft copolymer has been characterized in details using FTIR and 1H NMR
Received in revised form 17 July 2018 spectral analyses, GPC, TGA, FESEM, TEM and DLS analyses. GPC analysis results indicate that the polymerization
Accepted 31 July 2018
is controlled, while the LCST value of the copolymer suggests that the synthesized copolymer demonstrates sol-
Available online 1 August 2018
gel behaviour on applying temperature. FESEM and TEM analyses envisage that the graft copolymer has spherical
Keywords:
morphology with nanoscale dimension. DLS study reveals that the micellar size vary with change of pH and tem-
Amphiphilic perature, demonstrating the dual-responsive behaviour of the graft copolymer. Finally, the developed graft co-
RAFT polymerization polymer shows worthy efficacy towards the pH and thermo-responsive release of encapsulated pyrene in
Micelle sustained manner.
© 2018 Elsevier B.V. All rights reserved.

1. Introduction Various polymerization techniques have been used to synthesize

Recently, synthetic copolymers have received noteworthy attention
towards controlled release applications [1–3] along with potential can-
didates for tissue engineering [2]. Amongst synthetic polymers, amphi-
philic copolymers that form micelle are considered as the suitable
carriers because of their biocompatibility and high drug loading capacity
[3,4]. Recently, stimuli-responsive amphiphilic copolymers (that in-
clude response to temperature, pH, light, magnetic field, electric field)
have been focused with significant interest in biological science [2,3].
Temperature is an important stimulus in environmentally responsive
polymers and hence thermoresponsive systems are used in-vitro as
cell sheet manipulation methods [3], can also be utilized in-vivo using
the micelle containing drugs [5,6]. On the other hand, pH responsive
polymers can be used for targeted delivery. Henceforth, the combina-
tion of pH and temperature responsive amphiphilic copolymers may
change hydrophilic/hydrophobic balance, which makes a polymer
more suitable as targeted drug delivery system [7,8]. Recently, a dual-
responsive amphiphilic graft copolymer, composed of dextrin and
poly (N-vinyl caprolactam) has been synthesized by us, where dextrin
has been chosen owing to its easy availability, non-toxicity, economic
viability and high solubility [9,10]. On the other hand, N-vinyl caprolac-
tam (NVCL) has been used to graft on dextrin though reversible addition
fragmentation chain transfer polymerization (RAFT) that possesses am-
phiphilic character with hydrophilic amide group (lactam ring), at-
tached to the hydrophobic vinyl group [11].
⁎ Corresponding author.
E-mail address: sagarpal@iitism.ac.in (S. Pal).
amphiphilic graft copolymers. Conventional free radical polymerization
process is usually avoided because of its non-living character that re-
sults unwanted homopolymer [12]. Thus to overcome the disadvan-
tages of free radical polymerization process, controlled radical
polymerization (CRP) techniques are used for the fabrication of amphi-
philic copolymers. Specially, RAFT is the most preferable technique as it
can be used with variety of monomers and reaction conditions. As a re-
sult, it provides efficient control over molecular weight and polydisper-
sity (PDI) [13–15]. For RAFT polymerization, xanthate is used as RAFT
agent that form less stable radical adduct, which provides control mo-
lecular weight with narrow polydispersity [16,17].
Previously, many researchers have developed and reported NVCL
based graft copolymers [18–21] and block copolymers [22–25]. For ex-
ample, Rejinold et al., 2011, Eswaramma et al., 2017, Li et al., 2012,
Mogos et al., 2014, Hinkley et al., 2004 and Gois et al., 2016 have re-
ported the synthesis of NVCL based copolymer and their applications
as 5-fluorouracil carrier, anti-HIV drug carrier, pervaporation perfor-
mance for dehydration of caprolactam solution, burn dressing, LCST be-
haviour. The above mentioned reports did not discuss in details about
micellization behaviour. Though Gois et al., 2016 and Liang et al., 2015
have reported micellization behaviour, but the synthesized polymers
are block copolymers. Here dextrin and poly (N-vinyl caprolactam)
based amphiphilic graft copolymer has been synthesized through
RAFT polymerization that was used to encapsulate pyrene. Controlled
nature of the graft copolymerization has been investigated by GPC anal-
ysis. The critical micelle concentration (CMC) of the graft copolymer
was determined using fluorescence spectrophotometer, where pyrene
was used as probe molecule. Moreover, the stimuli-responsive

https://doi.org/10.1016/j.ijbiomac.2018.07.198
0141-8130/© 2018 Elsevier B.V. All rights reserved.
 A. Pal et al. / International Journal of Biological Macromolecules 119 (2018) 954–961 955

behaviour of the graft copolymeric micelle has been investigated by de-
termining LCST and hydrodynamic diameter. Finally, the copolymer
demonstrates worthy efficacy towards pH and temperature responsive
controlled release of encapsulated pyrene.
2. Experimental method
2.1. Reagents and materials
compound); 4.32 ppm (\\CH2, RAFT agent); 0.85 ppm (\\CH3, RAFT
agent); 1.67 ppm (\\CH2 of PNVCL); 4.03 ppm (\\CH of PNVCL);
2.73 ppm (\\CH2, H12, lactam ring); 1.54 ppm (\\CH2, H13, lactam
ring); 1.38 ppm (\\CH2, H14, lactam ring); 2.89 ppm (\\CH2, H15, lactam
ring)]. Different grades of the copolymer were synthesized with varia-
tion of reaction parameters/conditions and optimized (Table 1).
% Conversion was determined using Eq. (1) [26].

Dextrin and AIBN were procured from Loba Chemie, India. Pyridine, Wp
%Conversion ¼ ð1Þ
and N,N-dimethyl formamide were obtained from Merck, India. N-vinyl Wm
caprolactam and 2-bromo-2-methyl propionyl bromide (98% pure)
were received from TCI Chemicals, Japan. For experimental works, dou-
ble distilled water was used. Potassium ethyl xanthate was obtained Wp is the weight of dried graft copolymer and Wmis the initial weight
from Alfa–Aesar, USA. of monomer.

2.2. Synthesis 2.3. Characterizations

2.2.1. Synthesis of macroinitiator
At first, 0.5 g of dextrin [1H NMR (400 MHz, d6-DMSO) δ:
5.40–5.49 ppm (\\CH, H1), 3.06 ppm \\CH, H2 and H4); 3.64 ppm
(\\CH, H3); 3.58 ppm (\\CH, H5), 3.46 ppm (\\CH2, H6), 4.58 ppm
(\\OH2); 4.90 ppm to 5.10 ppm \\OH3 to \\OH6)] was dispersed in
mixture of pyridine (6 mL)/water (10 mL). The solution mixture was
heated at 80 °C until become a transparent solution. Then 2-bromo-2-
methyl propionyl bromide (2 mL) was added drop wise in cold condi-
tion. The reaction was continued at room temperature for 12 h with
continuous stirring. The resultant mixture was poured into excess
acetone, and precipitated. Afterward, the residue was dried under
vacuum at 55 °C for 2 days. The reaction was carried out in inert
atmosphere of nitrogen [1H NMR (400 MHz, d6-DMSO) δ: 3.04 ppm to
5.48 ppm (Hs and \\OHs of dextrin) and 1.24 ppm (\\CH3 of bromo-
ester compound)].
2.2.2. Development of g-Dex/PNVCL graft copolymer
The controlled synthesis of g-Dex/PNVCL was processed through
RAFT polymerization. 0.1 g of macroinitiator was dissolved in 25 mL
DMF in a 100 mL 3-necked RB flask. The RB was fixed with stirrer that
was kept on an oil bath with a constant temperature of 80 °C. Then
1.0 g (0.0062 mol) of potassium ethyl xanthate was incorporated to
the reaction mixture and the reaction was continued for 12 h. Then
the product was filtered using Whatman 41 filter paper and dried
under hot air oven at 55 °C for 72 h.
After that, 1.0 g of xanthate mediated dextrin compound was taken
in RB. Then 0.02 g of AIBN and 0.021 mol of NVCL were added sepa-
rately. Subsequently the reaction was performed at 75 °C for 3 h with
continuous nitrogen purging. The reaction temperature was brought
down to ambient and precipitated in 250 mL acetone. Finally, the col-
lected product was dissolved in acetone and water mixture (30:70). Af-
terward, it was heated to 60 °C and the product was separated out. This
process was repeated for 4 times to remove unreacted NVCL. The prod-
uct (i.e. graft copolymer) was dried in a hot air oven at 55 °C for 12 h [1H
NMR (400 MHz, d6-DMSO) δ: 1.10 ppm (\\CH3, bromo-ester
The synthesized copolymer has been characterized using various
techniques that include FTIR and 1H NMR spectroscopy, TGA analysis,
GPC, DLS, AFM, FESEM and TEM analyses. The instrumental details and
experimental procedure have been illustrated in Supporting
information.
2.4. Determination of lower critical solution temperature (LCST)
The LCST of graft copolymer was determined by optical transmit-
tance of the copolymer solution (25 mg/L) at a temperature range of
25 °C to 45 °C with Agilent Cary 5000 UV–vis spectrophotometer.
2.5. Determination of CMC of g-Dex/PNVCL graft copolymer
The CMC of the synthesized graft copolymer was measured using
fluorescence spectrophotometer (Model: LS55; Make: Perkin Elmer
USA).
2.6. Encapsulation of pyrene on copolymer and its release
The encapsulation of pyrene on copolymer and its release was per-
formed in the similar way as reported before [27]. The detailed proce-
dure has been explained in Supporting information.
3. Results and discussion
3.1. Synthesis
The proposed synthetic step for the development of graft copolymer
is demonstrated in Scheme 1. At first, the macroinitiator was prepared
using dextrin and 2-brmo-2-methyl propionyl bromide, which was
then reacted with potassium ethyl xanthate to form the xanthate medi-
ated dextrin. Finally, NVCL was grafted on xanthate mediated dextrin in
presence of AIBN to obtain g-Dex/PNVCL graft copolymer as proposed in
Scheme 1.

Table 1
Synthesis details, molecular weight and polydispersity of various g-Dex/PNVCL graft copolymers.

Compound Time (h) Monomer (mol) % Conversion Mn Mw Polydispersity index

(kDa) (kDa)

Dextrin – – – 5.8 8.0 1.39

g-Dex/PNVCL 1 2 0.007 38 65.2 93.8 1.43
g-Dex/PNVCL 2 3 0.007 45 84.0 101.6 1.20
g-Dex/PNVCL 3 3.5 0.007 40 70.3 88.2 1.25
g-Dex/PNVCL 4 3 0.014 53 88.4 103.4 1.17
g-Dex/PNVCL 5 3 0.021 64 149.7 166.8 1.11
g-Dex/PNVCL 6 3 0.024 58 140.7 159.1 1.13

Bold indicates the optimized grade among synthesized graft copolymers.

956 A. Pal et al. / International Journal of Biological Macromolecules 119 (2018) 954–961
Scheme 1. Schematic representation for the preparation of g-Dex/PNVCL.
3.2. Characterization
3.2.1. GPC analysis
The controlled nature of graft copolymerization was studied by GPC
analysis. It was found that with increase in % conversion, Mn was pro-
portionately increased while polydispersity was decreased (Fig. S1a,
Supporting information), suggesting the living nature of polymerization
[13,16]. The kinetics of the graft copolymerization is represented in
Fig. 1. The plot of ln ([M]0/[M]t) vs. time is linear and passed through
the origin (Fig. 1a), which indicates that the polymerization is first-
order with respect to monomer concentration [28]. It was also observed
that the elution time for g-Dex/PNVCL 5 copolymer was lower than dex-
trin (Fig. 1b) that confirms the higher molecular weight of the copoly-
mer (Fig. 1d) as compared to dextrin (Mn: 5.8 kDa; polydispersity
~1.39) (Fig. 1c). The molecular weight and polydispersity of various
graft copolymers are given in Table 1.
3.2.2. FTIR spectra
FTIR spectrum of dextrin (Fig. S2a, Supporting information) demon-
strates the peak at 3285 cm−1, which can be assigned to the stretching
vibrations of O\\H. Other peaks at 2923 cm−1 and 1411 cm−1 can be at-
tributed to the C\\H stretching and bending vibrations, respectively.
The peaks at 1011 cm−1 and 927 cm−1 can be assigned to C\\O\\C
stretching vibrations [29]. After esterification of dextrin, the new peak
was observed at 1718 cm−1, which is responsible for the C_O
stretching vibration of ester carbonyl of bromo ester group (Fig. S2b,
Supporting information). The peak at 682 cm−1 is due to the C\\Br
bond vibration. Fig. S2c, Supporting information represents the FTIR
spectrum of potassium ethyl xanthate. The two characteristic peaks of
xanthate compound were observed at 1043 cm−1 and 660 cm−1,
which are attributed to\\C_S and C\\S [23]. After incorporation of po-
tassium ethyl xanthate to macroinitiator, these two characteristics
peaks were also found with little shifting to 1060 cm−1 and 654 cm−1,
suggesting successful assimilation of xanthate group (Fig. S2d,
Supporting information). NVCL (Fig. S2e, Supporting information)
shows the peaks at 3319 cm−1, 2927 cm−1, 1658 cm−1, 1476 cm−1,
1427 cm−1, 3107 cm−1 and 963 cm−1, which are corresponding to
N\\H stretching vibration, aliphatic C\\H stretching vibration, C_O
stretching vibration, C\\N stretching vibration, \\CH2\\ stretching vi-
bration, _CH and _CH2 stretching vibrations, respectively. On the con-
trary, after graft copolymerization of PNVCL on dextrin, all the
characteristics peaks of NVCL were observed except the peaks responsi-
ble for _CH (3107 cm−1) and _CH2 (963 cm−1) (Fig. S2f, Supporting
information). The disappearance of these peaks indicates the polymeri-
zation of NVCL through vinylic double bond.
3.2.3. 1H NMR spectra
1
H NMR spectrum of dextrin is shown in Fig. S3, Supporting informa-
tion. It is apparent that H2, 4 peaks of dextrin were observed at δ =
3.06 ppm, while H3, 5, 6 peaks of dextrin were found at δ =
3.46–3.64 ppm. The hydroxyl protons of dextrin (\\OH2, \\OH3 and
\\OH6) were observed at δ = 4.58–5.10 ppm. Besides, the anomeric hy-
drogen was obtained at δ = 5.40–5.49 ppm.
 A. Pal et al. / International Journal of Biological Macromolecules 119 (2018) 954–961 957

Fig. 1. (a) GPC kinetics plot; (b) elution time vs RI plot of copolymer and dextrin. (c) GPC curves of dextrin and (d) g-Dex/PNVCL 5.

Fig. 2. 1H NMR spectrum of g-Dex/PNVCL 5.

958 A. Pal et al. / International Journal of Biological Macromolecules 119 (2018) 954–961
Fig. S4, Supporting information demonstrates the 1H NMR spectrum
of the macroinitiator. One new peak of methyl proton was appeared at δ
= 1.24 ppm (H7), suggesting incorporation of bromo-ester group onto
dextrin backbone. It is also believed that most of the hydroxyl groups
took part in the formation of macroinitiator [9].
1
H NMR spectrum of NVCL is shown in Fig. S5, Supporting information.
The \\CH2 proton peaks of lactam rings were observed at δ =
1.56–1.58 ppm (H13), δ = 1.48 ppm (H14), δ = 2.46–2.49 ppm (H12) and
δ = 3.51–3.54 ppm (H15). Moreover,\\CH2 (H10) and\\CH (H11) protons
were found at δ = 4.45–4.48 ppm and δ = 6.94–7.01 ppm, respectively.
Finally, the 1H NMR spectrum of g-Dex/PNVCL 5 is represented in
Fig. 2. The methyl (H9) and methylene peak (H8) of xanthate group
were found at δ = 0.85 ppm and δ = 4.32 ppm, correspondingly. Addi-
tionally, the methylene peak of PNVCL was found at δ = 1.67 ppm (H10),
methine peak was observed at δ = 4.03 ppm (H11). Also the proton
peaks of lactam ring were found at δ = 1.38 ppm (H14), δ = 1.54 ppm
(H13), δ = 2.73 ppm (H12), and δ = 2.89 ppm (H15). These characteris-
tics peaks confirm the successful grafting of NVCL on dextrin through
RAFT polymerization.
The grafting ratio of hydroxyl protons was determined using Eq. (2)
[29] and it has been observed that the grafting ratio of copolymer was
0.46 [where IH10 = 2.23 and I(−OH) = (2.04 + 2.78) = 4.82]
IH10 ðCopolymerÞ
%grafting ratio of hydroxyl groups ¼
I ð−OHÞ
We have also determined % conversion (using Eq. (3)) [30] by com-
paring 1H NMR spectra of integral value of NVCL and g-Dex/PNVCL 5
and it was found that % conversion was 61%.
IH10ðcopolymerÞ
%Conversion ¼
IH10 ðcopolymerÞ þ 1H10 ðNVCLÞ
3.2.4. Thermogravimetric analysis
TGA analyses of dextrin, macroinitiator and g-Dex/PNVCL 5 are
shown in Fig. S6, Supporting information. Dextrin (Fig. S6a) is character-
ized with two degradation zones. The first weight loss in the range of
50–120 °C is mainly because of the evaporation of water molecule. An-
other weight loss in the region of 250C–340 °C is responsible for the
degradation of polysaccharide backbone [9]. TGA plot of macroinitiator
is presented in Fig. S6b, Supporting information. One additional weight
loss zone was observed in the range of 300 °C–380 °C, which is mainly
due to the degradation of bromo compound [9]. The TGA plot of graft co-
polymer (g-Dex/PNVCL 5) is demonstrated in Fig. S6c, Supporting infor-
mation. Two additional degradation zones were found at 120–160 °C for
Fig. 3. Investigation of surface morphology of g-Dex/PNVCL 5 graft copolymer using FESEM, and TEM analyses.
thermal decomposition of RAFT agent [25] and another weight loss zone
(410–530 °C) was responsible for the decomposition of PNVCL [31].
3.2.5. FESEM, AFM, and TEM analyses
It was found that dextrin has granular morphology [9]. After grafting
of NVCL on dextrin through RAFT polymerization, the graft copolymer
demonstrates spherical topography/morphology in nanoscale dimen-
sion as evidenced from AFM (Fig. S7, Supporting information), FESEM
(Fig. 3a), and TEM (Fig. 3b) analyses. Here it is worth to mention that di-
ameter of the copolymer is slightly higher as obtained from DLS analy-
sis, which is mainly because the DLS study was performed in the
aqueous phase [32].
3.2.6. pH- and thermo responsive behaviour of g-Dex/PNVCL 5 copolymer
LCST is a critical parameter to explain the temperature responsive
behaviour of a polymer. It has been observed from Fig. 4a that ~50% re-
duction of transmittance was revealed at 35 °C, which is corresponding
to the LCST of synthesized graft copolymer. Fig. 4b shows the hydrody-
namic diameter (DH) of the graft copolymer with reference to tempera-
ture. It is apparent that below LCST (i.e. at 25 °C), the DH value was quite
high than that of above LCST. This is because, below LCST (at 25 °C), H-
bonding interactions were predominated between amide group of lac-
tam ring and H2O, resulting higher hydrodynamic diameter (i.e.
346.0 nm). On the contrary at 37 °C, phase separation has been taken
ð2Þ
place and hence the hydrophobic interaction was dominated [23],
resulting lower hydrodynamic diameter of g-Dex/PNVCL 5
(246.5 nm). Fig. 4c demonstrates the variation of DH with respect to
pH. It was found that at pH 1.2, the DH value was 204.4 nm, while at
pH 7.4 the value was 383.3 nm. This is obvious because in acidic condi-
tion, mostly the hydrophilic groups were protonated that reduced the
ability to form of H-bonding with the media as compared to alkaline
ð3Þ
media. The above observations thus authenticate the dual responsive
(pH and thermo) behaviour of the synthesized graft copolymer.
3.2.7. Determination of CMC of g-Dex/PNVCL 5 copolymer and its micellar
stability
CMC of the synthesized graft copolymer was determined using fluo-
rescence spectroscopy at 37 °C where pyrene was used as fluorescence
probe. It was reported before that although pyrene has five characteris-
tics peaks, however, first (I1) and third (I3) peaks are considered be-
cause of its sensitivity towards polarity of microenvironment, where
peaks are located [32, 33]. Fig. 5a shows the variation of I1/I3 ratio
with graft copolymer concentration. It has been revealed that two linear
lines were intersected at 0.25 mg/mL, which is the CMC of the synthe-
sized copolymer. It is uncertain whether CMC was really formed at
0.25 mg/mL or not. Therefore, we had performed DLS study for three
 A. Pal et al. / International Journal of Biological Macromolecules 119 (2018) 954–961
Fig. 4. (a) Temperature dependence optical transmittance, (b) hydrodynamic diameter of copolymer at 25 °C and 37 °C, and (c) hydrodynamic diameter of copolymer at pH 1.2 and 7.4.
different polymer concentrations (0.15 mg/mL, 0.25 mg/mL and
0.50 mg/mL) at 37 °C (Fig. 5b). It is obvious that as the concentration
was increased, DH value was also increased (203.9 nm for
0.15 mg/mL; 246.5 nm for 0.25 mg/mL and 289.3 nm for 0.50 mg/mL)
with unimodal distribution. This DLS data indicate that more concen-
trated solution promote more aggregation between particles, resulting
higher DH [23]. However, it has also been observed that by dissolving
0.25 mg/mL of copolymer, a comparatively narrow PI (~0.546) with
perfect monodisperse distribution was found. Therefore, it may be
considered that at 0.25 mg/mL, the hydrophilic segments offer a stable
interface between the hydrophobic part (PNVCL) and the aqueous me-
dium, which form more stable self-assembly structures [23] than the
others.
The micellar stability of the developed graft copolymer has been in-
vestigated by DLS analysis for 5 days (Fig. 5c and Fig. S8, Supporting in-
formation). It was revealed that there was no significant change of
hydrodynamic diameter (the value remains in between 240 nm–
250 nm) during the investigated period (Fig. 5c), signifying long term
stability of the graft copolymer micelle [31].
3.2.8. Encapsulation and dual responsive release behaviour of copolymer
After pyrene encapsulation (Fig. S9, Supporting information), the
pH and temperature stimulated release of pyrene from the graft
Fig. 5. (a) Critical micelle concentration of copolymer; (b) hydrodynamic diameter of graft copolymer with different concentrations; and (c) hydrodynamic diameters of graft copolymer
with time (day).
959
copolymer was performed for 96 h. UV–vis spectra were recorded
to discern how much amount of dye was released from core of the
micelle. It has been noticed (Fig. 6a) that at pH 7.4, % release was
higher as compared to that of pH 1.2. This can be attributed by the
fact that in basic condition, the hydrophilic groups of graft copoly-
mer chains would be able to interact easily with the media, leading
to higher % dye release [9,10]. Moreover the release behaviour was
also carried out at two different temperatures (Fig. 6b). It is obvious
that the release rate was slower beyond LCST. This can be explained
based on the fact that above LCST, hydrophobic nature is
predominating that leads to coil the structure of the copolymer,
which slower the release rate [29].
4. Conclusion
Dual responsive, amphiphilic graft copolymer based micelle has
been successfully synthesized by RAFT polymerization. Kinetics study
reveals that graft copolymerization is first-order with respect to mono-
mer. 1H NMR spectral analysis confirms the formation of copolymer.
Temperature responsive nature of the copolymer was assured by deter-
mining LCST, which was well supported by DLS data. It was also found
that there was no major change in hydrodynamic diameter during the
investigated period (for 5 days), which signify the long term stability
960 A. Pal et al. / International Journal of Biological Macromolecules 119 (2018) 954–961
Fig. 6. (a) % Release of pyrene from copolymer micelle as a function of pH; (b) % release of pyrene from copolymer micelle as a function of temperature.
of copolymer micelle. Finally, the release of encapsulated pyrene dem-
onstrates that this new graft copolymeric micelle showed excellent
dual-responsive controlled release behaviour.
Acknowledgement
Authors thankfully acknowledge the financial support from Indian
Institute of Technology (ISM), Dhanbad, India as well as the various in-
strumental facilities of CRF, Indian Institute of Technology (ISM),
Dhanbad, India.
Appendix A. Supplementary data
Detail characterization techniques; experimental procedure for
pyrene encapsulation and release; GPC analysis (Fig. S1); FTIR
spectra (Fig. S2); 1 H NMR spectrum (Figs. S3, S4 and S5); TGA
analysis (Fig. S6); AFM analysis (Fig. S7); DLS study (Fig. S8);
pyrene encapsulation study (Fig. S9). Supplementary data to this
article can be found online at https://doi.org/10.1016/j.ijbiomac.
2018.07.198.
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