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Current Environmental Engineering, 2019, 6, 134-140

RESEARCH ARTICLE
ISSN: 2212-7178
eISSN: 2212-7186

Development of Graphene Oxide-Trihexyphenidyl Hydrochloride

Nanohybrid and Release behavior

Pradip M. Jawanjala, Pritam B. Patilb, Jayesh Patila, Mrunal Waghuldea and Jietndra B. Naika,*
a
University Institute of Chemical Technology, KBC North Maharashtra University, Jalgaon, 425 001, Maha-
rashtra, India; bDepartment of Chemical Engineering, Shri S'ad Vidya Mandal Institute of Technology,
Bharuch 392001, Gujarat, India

Abstract: Background: The demand of an efficient nanocarrier in drug delivery, graphene
and its derivatives are emerging as a rising star due to its remarkable chemical and struc-
tural properties.
Objective: Graphene oxide (GO) has high surface area and ability to load high amount of
aromatic drugs. Hence, the objective of the research was to load Trihexyphenidyl hydro-
chloride (THP), antiparkinsonian drug on GO ultrasonically by π-π stacking interaction.
Current Environmental Engineering

A R T I C L E H I S T O R Y

Methods: GO was synthesized by the modified Hummer method. The conjugation of GO-
Received: November 29, 2018 THP was generated by using Design-Expert Software and release study of GO-THP nano-
Revised: February 1, 2019
Accepted: March 8, 2019
hybrids was performed in the dissolution tester by using a dialysis membrane.
DOI:
Results: By varying an amount of GO and THP, the effect on loading efficiency and drug
10.2174/2212717806666190313153239 release was studied. THP showed sustained release behavior with release efficiency of
 89% to 98% over 8 h. GO-THP complex was characterized by UV-vis spectrophotometer,
FTIR, FESEM and XRD analysis.
Conclusion: GO-THP complex showed better-sustained release of the drug and can be useful
for the reduction dose frequency as well as adverse effect with better patient compliance.
Keywords: Graphene oxide, trihexyphenidyl hydrochloride, parkinson's, nanocarrier, nanohybrid, FTIR.
1. INTRODUCTION including its high specific surface area, two-
In drug delivery, nanomaterials ranging from 1- dimensional structure, and biocompatibility. Due
100 nm are used as transport modules such as mi- to these properties GO is considered as an ideal ma-
celles, polymers, carbon-materials, liposome, den- terial for a wide range of applications in the medi-
drimers, etc. [1, 2]. Due to remarkable chemical cal field such as drug or gene delivery, biosensor,
and structural properties, graphene-based materials medical imaging, tissue engineering, photothermal
are widely used in drug delivery application in the therapy, and antibacterial applications [13-18].
past few years. GO has a single-layered carbon In addition, the plenty of π electrons available
atom bonded through sp2 hybridization, arranged on the surface of GO enable the π - π interaction
as a two-dimensional honeycomb lattice [3-6]. GO with aromatic drug compounds [7]. Therefore, GO
is an oxidized form of graphene, laced with oxy- can serve as an admirable drug delivery platform.
gen-containing functional groups such as carboxyl, Moreover, the amphiphilic property of GO allows
epoxyl, carbonyl, hydroxyl, etc. which occupies it to stabilize the hydrophobic molecules [15]. The
the 60% of total graphene oxide area [7-11]. Gra- drug-loading ratio of graphene nanomaterials
phene has a theoretical specific surface area of might reach to 200%, which is comparatively high-
2600 m2/g, which is four times higher than any er than any other nanomaterial and drug delivery
other material used for drug delivery [12, 13]. GO systems [13]. Graphene can be functionalized by a
possesses remarkable physiochemical properties number of materials to increase its properties such as
dispersibility or water solubility which may increase
*Address correspondence to this author at the University Institute biocompatibility and reduce its toxicity [1, 19].
of Chemical Technology, KBC North Maharashtra University,
Umavi Nagar, Post Box No. 80, Jalgaon-425 001, Maharashtra, India; GO-based drug delivery systems are mostly pH
Tel: +91257 2257441; Fax: +91257 2258403;
E-mail: jbnaik@nmu.ac.in sensitive, notably in alkaline and neutral state, the

2212-7186/19 $58.00+.00 © 2019 Bentham Science Publishers
Development Of Graphene Oxide
nature of drug release is slower, thus signifying a
sustained drug delivery system [15]. Moreover,
GO has the capability to deliver drugs at a specific
(targeted) site by sensing temperature, pH and
photo thermally, electrically and magnetically con-
trolled external agents [7, 20]. GO layers and their
thickness play a key role for numerous reasons
such as many layers will reduce the surface area
but will increase the rigidity of graphene nanocar-
riers required for cell penetration. In addition, the
lateral dimension of GO does not influence surface
area and drug loading [13]. To decrease the dosing
frequency and adverse effect, sustained formula-
tion of a drug is indispensable [21, 22]. THP is
antidyskinetic and antiparkinsonian drug whose
IUPAC name is 1-cyclohexyl-1-phenyl-3-(1-
piperidyl-1-propanol1) [23]. THP is used for the
symptomatic treatment of Parkinson’s disease in
mono and combination therapy [24].
Herein, we report a non-covalent nanohybrid
formed by THP with GO and investigated it for the
loading and in vitro release of THP by GO
nanosheets.
2. MATERIALS AND METHODS
2.1. Materials
Graphite powder (100 micron) was purchased
from SDFCL (S. D. Fine-Chem Limited). 85%
pure Ortho-acid and Hydrogen peroxide 30% was
purchased from MERCK (Merck specialties pri-
vate limited). Trihexyphenidyl hydrochloride was
obtained as a gift sample from S. P. Pharmaceuticals,
Jalgaon (Maharashtra). Other chemicals and reagents
were of analytical grade and used as received.
2.2. Synthesis of Graphene Oxide
Graphite flakes can be oxidized using three dif-
ferent procedures: improved method, Hummers’
method, and Modified Hummers’ method with ad-
ditional KMnO4 [10, 25].
In modified Hummer's method, graphite flakes
and NaNO3 are added to1000ml volume flask con-
taining H2SO4 below the 5°C temperature with
continuous stirring. H2SO4 exfoliates the number
of graphite layers from graphite flakes. The mixture
was stirred for 1 h and potassium permanganate
was added into the suspension very slowly with an
operating temperature lower than 15°C. The
mixture was diluted with the very slow addition of
water and kept under stirring for 2 h at 35°C. The
above mixture was kept in a reflux system at 98°C
Current Environmental Engineering, 2019, Vol. 6, No. 2 135
for 10-15 min. Then, the temperature was
maintained at 25oC for 2 h. The solution was
finally treated with H2O2 to complete the reaction
and the colour of the solution changed to bright
yellow. Then the suspension was diluted by adding
water. The resulted mixture was washed
repeatedly with 10% HCl and subsequently with
deionised (DI) water several times. After washing,
the gel like substance was vacuum dried at 60°C
for more than 6 h to get dry GO powder [26].
2.3. Conjugation of THP and GO
The conjugation of GO-THP batches was gener-
ated by using Design-Expert Software (Stat-Ease
Inc., Minneapolis) to get an optimum batch [27-29].
Firstly, GO was sonicated with THP in methanol
for 1 h and then stirred overnight at room tempera-
ture in the dark condition [19]. The dry composition
remains at the bottom, after evaporation of metha-
nol. This composition was washed repeatedly with
distilled water to remove the unbound drug and
centrifuged at 1000 RPM for 15 min. Then the
product was freeze-dried to obtain a final conjuga-
tion of GO-THP. The resultant product of GO-THP
was characterized by UV-Vis spectrophotometer,
FT-IR spectrometer, FESEM and X-ray diffraction.
2.4. Loading of THP
Loading of THP was studied by separating the
GO-THP nanohybrid from aqueous medium con-
taining free THP molecules or unloaded THP. The
unloaded THP was measured by UV-vis spectro-
photometer at 218 nm. For this, a standard calibra-
tion curve was constructed by a series of THP so-
lutions with different concentrations.
2.5. Release of THP from GO-THP
Release study of GO-THP nanohybrids was
performed in the Electro lab dissolution tester by
using a dialysis membrane. GO-THP nanohybrids
containing 5 mg THP, equivalent to the dose, dis-
persed in 5 ml aqueous solution and put into the
dialysis membrane. Dialysis membrane was placed
in the dissolution tester, which contains a buffer
phosphate solution of pH-6.8. The release reser-
voir was kept under constant stirring at 37˚C±
0.5˚C. Samples were withdrawn at regular time
intervals and replenished with dissolution media to
maintain the sink condition. The concentration of
THP released from GO-THP into the buffer solu-
tion was quantified by using UV-Visible spectros-
copy [27, 30].
136 Current Environmental Engineering, 2019, Vol. 6, No. 2
3. RESULTS AND DISCUSSION
The -OH and -NH groups present on THP can
form a strong hydrogen bonding interaction with
-OH and -COOH groups present on GO sheet. It
means THP can simply load on GO noncovalently
with the aid of slight sonication.
SEM image of GO shows nanosheets laying
one over another with smooth surface shown in
Fig. (1a) while in Fig. (1b) nanohybrids observed
a large amount of THP loaded on GO
nanosheet.
S-4800 15.0kV 13.7mm x20.0k SE(M)
Fig. (1a). SEM image of GO.
S-4800 15.0kV 13.6mm x20.0k SE(M)
Fig. (1b). SEM image of GO-THP.
Jawanjal et al.
The more confirmation came from absorbance
peaks of GO, THP and GO-THP by UV-vis spec-
troscopy shown in Fig. (2a). The Go has single
broad peak at 204 nm, and THP solution absorbs
strongly at 218, 241, 251 and 257 nm. The loading
of THP onto GO was evident from the spectrum of
the GO-THP nanohybrid solution, which shows a
characteristic absorption peak of THP clearly. Af-
ter forming the GO-THP nanohybrid, the absorp-
tion peaks of THP showed a slight-shift. The ab-
sorption peak of THP at 218nm shifted to 206nm
after hybridization with GO.
2.00mm
2.00mm
Development Of Graphene Oxide Current Environmental Engineering, 2019, Vol. 6, No. 2 137

218

THP
204
Absorbance

GO

206

GO-THP

200 250 300 350

Wavelength
Fig. (2a). UV-vis spectra of THP, GO, and GO-THP.

GO
Transmittance

THP

GO-THP
4000 3500 3000 2500 2000 1500 1000 500
Wave Number (cm-1)
Fig. (2b). FT-IR spectra of GO, THP and GO-THP.

FT-IR spectra of GO, THP and GO-THP nano-
hybrid recorded in the wavelength region of 4000-
400 cm-1are shown in Fig. (2b). GO shows O-H
stretching vibration at 3321cm-1, C = O carbonyl
stretching at 1731 cm-1, C-OH at 1224 cm-1 and C-
O at 1053 cm-1. The peak at 1731cm-1 corresponding
to C = O in the spectrum of GO and the Peak at
1728cm-1 in the spectrum of the THP shift to lower
at the 1716 cm-1 after forming GO-THP nanohybrid.
This may show that THP loaded onto GO, and the
shift of the characteristic peak may be due to hydro-
gen bonding between the two components [19].
138 Current Environmental Engineering, 2019, Vol. 6, No. 2 Jawanjal et al.

12.075

GO
16.75
Counts

THP
15.20

GO-THP
10 20 30 40 50 60 70 80

2 Theta (Coupled 2 Theta/Theta)WL=1.5406

Fig. (3). X-ray diffraction pattern of GO, THP, and GO-THP.

Physical nature of GO nanosheet, THP and GO- shows shifting in diffraction angle at 2θ =15.20˚
THP nanohybrid were examined by X-ray diffrac- and amorphous nature of GO reduced from 53% to
tometer shown in Fig. (3). XRD analysis showed that 45%, that means it confirms the majority of THP
the GO nanosheets are 53% amorphous in nature. loaded homogeneously onto GO nanosheet.
The sharp diffraction peak observed at 2θ = 12.075˚ The loading of THP on GO was investigated by
corresponds to GO [26]. XRD analysis corre- using equation obtained from the calibration curve
sponds to THP shows diffraction 2θ at 16.75˚ and formed by a series of THP solutions with different
it shows crystalline nature. GO-THP nanohybrid concentrations. The composition of GO varies in
(a)
45
40
35
30
Loading In %

25
20
15
10
5
0
04:03 03:02 02:01 08:03

GO:TH

Fig. (4a). % THP loading on GO of all experimental runs.

Development Of Graphene Oxide Current Environmental Engineering, 2019, Vol. 6, No. 2 139

(b)
120

100

80
DRUG RELEASE (%)

04:03

60 03:02

02:01
40
08.03

20

0
0 2 4 6 8 10
HOUR

Fig. (4b). Cumulative % drug release from GO-THP nanohybrids of all experimental runs.
between 100 to 200 mg and THP in between 50 to THP and GO was mainly due to the strong π -π
100mg. The interaction between GO and THP is stacking interaction. In XRD analysis, we have
attributed to π -π stacking interaction. The hydro- observed that the amorphous nature of GO reduced
gen bond between the -OH and -NH groups of from 53% to 45%, which confirms the loading of
THP and -OH and -COOH groups of GO may ex- THP on GO. THP showed sustained release be-
ist. It was observed that the loading of THP onto havior with the release efficiency of 89% to 98%
GO increases with the increase of GO in propor- over the period of 8 h. Thus, it can be concluded
tion to THP. GO and THP in the proportion of 4:3 that the GO is the convincing carrier for sustained
the loading was found to be 23%. By increasing release of THP.
the proportion of GO from 4:3 to 8:3; 40% loading
was obtained. It is due to the increase of the GO LIST OF ABBREVIATIONS
surface area for loading THP, which in turn in-
creases the loading efficiency shown in Fig. (4a). GO = Graphene Oxide
In view of the loading capacity of GO toward THP = Trihexyphenidyl Hydrochloride
THP, it may be carrier candidate. Fig. (4b)
showed the sustained release behavior of THP and FTIR = Fourier Transform-infrared Spec-
89% to 98% of drug release efficiency over the troscopy
period of 8h. It might be due to the strong hy- FESEM = Field Emission Scanning Electron
drogen bonding, which requires more time to Microscopy
release the drug.
XRD = X-Ray Diffraction Analysis
CONCLUSION
In this study, the loading of THP on GO and CONSENT FOR PUBLICATION
sustained release behavior of THP were investi- Not applicable.
gated. The loading of THP on GO increased with
increase in the GO surface area that is originated AVAILABILITY OF DATA AND
within the suspension, as multilayer GO MATERIALS
nanosheets were exfoliated. The bonding between Not applicable.
140 Current Environmental Engineering, 2019, Vol. 6, No. 2
FUNDING
Technical Education Quality Improvement
Program (TEQIP-III), World Bank and MHRD,
New Delhi, India.
CONFLICT OF INTEREST
The authors declare no conflict of interest,
financial or otherwise.
ACKNOWLEDGEMENTS
The authors are grateful to the Technical Edu-
cation Quality Improvement Program (TEQIP-II)
for providing financial assistance to carry out this
research work. Authors are also thankful to S. P.
Pharmaceutical, Jalgaon for providing a gift sam-
ple of Trihexyphenidyl hydrochloride.
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