Journal Review: Green Synthesis Nanoparticles for Glucose Biosensor

Applications
Nanang Fadillah_183200201, Pipit Fitria Yuliadi_11832005 1,
Reni Silvia Dewi_1183200241, Salsabila Regina Mayanti_1183200121

1
Engineering Physics, Institut Teknologi Sumatera

ABSTRACT
Nanoparticles are used in many industrial applications. The role of ZnO nanoparticles (ZnO
NPs) has various properties such as UV filter properties and photochemical activity, antifungal,
high catalyst, and antimicrobial. Due to the high levels of toxic chemicals and the extreme
environment used in chemical and physical methods, green synthesis has been adopted using
plants, fungi, bacteria, and algae for the synthesis of nanoparticles. This journal discusses how
the green synthesis in the formation of ZnO nanoparticles is then applied to glucose biosensors.
This journal is compiled using various journal sources to compile relevant review articles. From
several journals, the green synthesis is effective in the formation of ZnO nanoparticles and is
more environmentally friendly when compared to physical and chemical synthesis methods. On
the other hand, glucose biosensors prepared from several studies show good electrocatalytic
abilities for glucose determination, in addition to using the green synthesis ZnO NPs can increase
the sensitivity of the biosensor.
Keywords: Glucose Biosensor, Green Synthesis, Nanoparticles, Zinc Oxide.
Table of Contents

Introduction 1
ZnO Nanoparticles 2
Green Synthesis Method of ZnO Nanoparticles 5
Results and Discussion 6
Conclusions 8
Acknowledgments 8
References 8
Appendix 10

1. Introduction

Nanotechnology is a branch of science that is of great interest to researchers. Apart from its
potential applications that can be applied in various fields, nanotechnology is a technique that
includes the design, synthesis and characterization as well as the application of materials
organized in one dimension at the nanometer scale (100 nm in size) [1]. Nanoparticles are a class
of materials that includes particulate matter having one dimension less than 100 nm [2]. The size
of the nanoparticles depends on the constituent material, this material can be 0D, 1D, 2D or 3D.

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 1

The importance of this material makes researchers know that size can affect physiochemical
properties of a substance such as optical properties. Nanoparticles exhibit color characteristics
and properties with variations in size and shape, which can be utilized in bioimaging applications.
Changes in one of the factors that affect the absorption properties of nanoparticles [4].
Nanoparticles consist of three layers namely the surface layer, which can be functionalized
with various small molecules, metal ions, surfactants and polymers, the shell layer which is
chemically different from the core in all aspects, and the core, which is the central part of the
nanoparticle and usually refers to the nanoparticles. on the nanoparticles themselves [7].
One of the nanoparticles that are quite popular at this time is zinc oxide (ZnO) which has
exclusive properties such as semiconductor, piezoelectric, and optics so that this material can be
applied in various fields such as energy storage, nanosensors, cosmetic products, nano-optical
devices, electronic devices. nanoelectronics, and so on [6]. ZnO nanoparticles have attracted
much attention for the fabrication of efficient biosensors due to their unique features which
include high catalytic efficiency, high isoelectric point, non-toxicity, biocompatibility, excellent
electron transferability, and good stability. Some researchers are interested in using the green
synthesis for the synthesis of ZnO nanoparticles because the process is environmentally friendly,
cost-effective, and safe, it is also because plant extract-mediated nanoparticle biosynthesis is
economically advantageous and offers natural anchoring agents in the form of proteins.
One of the most popular biosensors, the glucose biosensor, has been studied extensively for
its importance in the food, environmental, and clinical industries. Glucose oxidation-based
amperometric biosensors (GOx) are now widely used for glucose monitoring due to their
reliability, low cost and simplicity. GOx is a flavoprotein that catalyzes the oxidation of
-D-glucose to hydrogen peroxide (H2O2)andglucono-1,5-lactone [1]. For the biosensor to function,
the GOx enzyme must be immobilized to the electrode surface to increase the durability and
stability of the enzyme; however, the immobilized matrix must be biocompatible and must not
interfere with the enzyme structure. Immobilization of enzymes on nanostructures facilitates
direct electron transfer between enzyme active sites and electrodes, formation of the desired
microenvironment, and creation of a large surface area for larger enzyme fabrication. Therefore,
the use of ZnO NPs material is very suitable to be immobilized to the electrode surface to
increase its electron transfer ability. In this journal review, we will briefly discuss the green
synthesis of ZnO NPs with various methods. In addition, it will also be explained related to the
results of the characterization of ZnO NPs samples and also the electrocatalytic properties of the
glucose biosensor made.

2. ZnO nanoparticles
In today's era, ZnO is widely used in various applications in all fields, such as sensors,
photocatalysts, piezoelectric, electrical circuits and so on. This compound has various advantages
compared to other oxide compounds. In recent years, ZnO is more likely to be applied in
biomedical, optical, and electronic fields [5] .
In addition to being registered in GRAS ( Generally Recognized As Safe) by theFood and
Drug Administration),ZnO NPs can improve food quality and food packaging when applied to the
food preservation and packaging industry as an antimicrobial agent. Packaging with antimicrobial
technology has the ability to reduce and inhibit the growth of bacteria on the packaging. This
application is carried out through three mechanisms, namely the release of antimicrobial ions, the
destruction of bacterial cells, and the formation of ROS using light radiation.

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 2

 Nanoparticles can be synthesized by various methods. However, in its application to the
field of biosensors, two main methods can be used, namely mechanochemistry (Mecano
Chemical Process) and physical vapor synthesis (Physical Vapor Deposition) [5].

2.1 Mechanochemical (MCP)

The MCP method is relatively new among other nanoparticle synthesis methods. The way
this method works is by combining a physical size reduction process in a ball mill using a
nanoscale chemical reaction during the milling process.
In this process, the precursor is milled in a ball mill to produce the desired compound by
the pounding process and chemical reaction [8].

Figure 2.1 MCP Process ZnO Nanoparticles [8]

Ball mill acts as a chemical reactor at low temperatures where the reaction can be produced at
room temperature and pressure. The resulting nanoparticles averaged between 20-30 nm,
depending on the time and temperature used during the ball milling. The longer the grinding
process, the smaller the size of the nanoparticles produced. The most optimal grinding time is 40
minutes. High and low temperatures can also affect the crystal size of the resulting nanoparticles.
The higher the temperature used, the size of the nanoparticles will also increase.
This MCP method is very beneficial if you want to synthesize ZnO nanoparticles on a large
(industrial) scale because the process is simple and the costs tend to be low. In addition, this
method is also considered environmentally friendly because the reaction involved is not an
organic solvent [8].

2.2 Physical Vapor Deposition

In this method, reactant gas is used which is then added to the vapor and then cooled to a
certain temperature and condensed to produce nanoparticles.

Figure 2.2 PVD ZnO Nanoparticles [8]

The nanoparticles produced in the PVD process consist of discrete (very dense) particles with

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 3

defined crystallinity. The nanoparticle size ranges from 8-75 nm.
In addition to the two main methods above, there are several other methods, namely
precipitation, thermal decomposition, and hydrothermal. The characteristics of this method for the
precursors and solvents used can be seen in Table 1.

Table 1. Synthesis Method of ZnO Nanoparticles [8]

Method Precursor Solvent Size (nm) Form

Coprecipitation Zinc acetate Distilled water 80 (length), 30-60 Nanorod

(diameter)

Microwave Zinc acetate dihydrate 1-Butyl-3-methylimid 37-47 Sphere

decomposition azolium bis
(trifluoromethylsulfon
yl) imide
[bmim][NTf2]

Hydrothermal Zinc acetate dihydrate Polyvinylpyrrolidone 5 (length), 20-300 Nanorod

(PVP) (diameter)

Wet chemical Zinc nitrate Sodium hydroxide 20-30 Acicular

hexahydrate (NaOH) as precursor
and soluble starch as
stabilizer

Sol-gel Zinc nitrate Distilled water and 30-60 Circular and

gelatin as substrate hexagonal

The process of synthesizing nanoparticles can be said to be quite difficult considering the many
variables that can affect the properties the resulting product. For the synthesis of ZnO
nanoparticles, it is very important to control the particle size in order to produce the same size. On
the other hand, the morphology of the nanoparticles is another important factor that must also be
considered, because it can affect the level of toxicity [8].
3. Method Green Synthesis ZnO Nanoparticles
Green Synthesis of ZnO nanoparticles is currently in great demand because of its safe,
environmentally friendly nature, ease of manufacture, and low production costs. Biosensors are
one of the most popular applications.
3.1 Coprecipitation Method
biosensors amperometric (GOx) are currently widely used for glucose monitoring because
of their low cost, low cost and simplicity. GOx is a flavoprotein that catalyzes the oxidation of
D-glucose to hydrogen peroxide (H2O2) and D-glucono 1,5 lactone. For the biosensor to function,
the GOx enzyme must be immobilized to the electrode surface to increase the durability and
stability of the enzyme. However, the immobilized matrix must be biocompatible and must not
interfere with the enzyme structure.
For the manufacturing process using root extract ingredients from Zingiber Officinale,
Glucose oxidase, graphite powder, and Zinc acetate dihydrate. The synthesis stage was carried
out using the precipitation method, namely Zinc Acetate was dissolved in 50 ml of double

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 4

distilled water to give a final concentration of 0.02M. Then, 20 ml of Zingiber Officinale (0.25
g/ml with distilled water) was slowly added to the Zinc acetate. This mixture was heated at 80 °C
and stirred for about 2 hours, until a white precipitate formed. The precipitated ZnO nanoparticles
were centrifuged for 10 minutes at 10,000 rpm, washed with distilled water, and then washed with
methanol to remove unwanted impurities. The ZnO nanoparticles were dried (at 60 °C overnight)
and then calcined at 450 °C in a furnace for 2 h [10]. Another researcher [11] used neem (A.
indica) leaf extract for the manufacture of ZnO NPs.synthesis process also uses the
coprecipitation method where neem leaf extract is mixed with 90 mL of zinc nitrate dropwise
under continuous stirring at room temperature for 4-5 hours. [5] using the same method also
synthesized ZnO NPs by adding Srigading Flower extract.
3.2 Hydrothermal
Method The hydrothermal method is a process that uses a single-phase or heterogeneous
reaction in aqueous solution at high temperature (T > 25 °C) and pressure > 100 kPa to crystallize
ceramic materials directly from solution. [12] using peach extract, the synthesis process using the
hydrothermal method. In the ZnO NPs synthesis procedure Znpowder was added to 30 mL of
peach juice, followed by 1 mL of ammonia solution in a Teflon Lined stainless steel autoclave
with a volume capacity of 50 mL. Then, the autoclave was tightly closed and maintained at 180
°C for 12 hours. After hydrothermal treatment, the autoclave was naturally cooled to room
temperature. The resulting product was filtered and washed with DI water several times, then the
residue was dried at 70 °C overnight. The completely dry powder sample was thenstored.
3.3 Sol-Gel
Method The sol-gel method is a method for synthesizing nanoparticles that applies two
phases , namely sol and gel. Sol is a colloidal suspension in which phase is a solid which is still
undergoing Brownian motion (Brownian motion) or Brownian diffusion (Brownian diffusion) and
the dispersion is a liquid. While the gel is a substance that has semirigid pores consisting of a
continuous network in three dimensions formed from polymer chains.
[13] In the biosynthesis process of ZnO nanoparticles using Citrus aurantifolia extract.
Citrus aurantifolia fruit that has been peeled, cut into small pieces and mixed with deionized
water. Puree until it becomes mush, then filtered. Use a muslin cloth to remove any solid
particles. After becoming a liquid, then filtered using a filter syringe with a pore size of 200 nm.
Zinc acetate is dissolved in 100 ml of solution. The liquid extract of C. aurantifolia was filtered
based on the concentrations of 0.05 M, 0.10 M, 0.15 M and 0.20 M. Its pH value was between 3.7
and 4.0. The mixture is then heated to 90° C. with continuous stirring for 3 hours until a white
precipitate appears. The precipitate was washed with deionized water and dried at 100°C for 6
hours.
4. Results and Discussion
4.1 Characterization of ZnO NPs
Synthesized ZnO nanoparticles can be analyzed by various characterization methods such
as XRD (X-Ray Diffraction), UV-vis Spectroscopic and scanning electron microscopy (SEM). In
research [10], UV-vis testing was carried out on ZnO NPs from ginger root extract. From the test
results, the absorbance peak was found at a wavelength of 365 nm which confirmed the formation

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 5

of ZnO nanoparticles. [5] also carried out UV-vis testing on the sample and obtained the
absorbance peak at a wavelength of 365 nm.
In addition, XRD testing was also carried out to see the crystallinity structure of ZnO NPs.
From the test results it was observed that the diffraction peak was highest at 2 theta angle which
was at 36.24 degrees which corresponds to the 101 lattice plane. The peak has been associated
with the hexagonal ZnO phase (JCPDS file: 36-1451). Then to determine the particle size of ZnO
NPs, calculations were carried out using the Debye Scherrer and obtained the particle size of
16.58 nm which was in the size range of 12-32 nm. [12] also carried out characterization using
XRD on the sample and obtained a high hexagonal crystal structure with a crystal size of 30 nm
according to the width of the plane of refraction line (101). [11] also reported his test which
resulted in 3 highest peaks at an angle of 2θ namely at 31.66°, 34.58°, and 36.54°. This line of
reflection corresponds to the wurtzite structure confirming the successful synthesis of ZnO NPs.
However, [13] obtained an XRD pattern of the structure of ZnO NPs with the highest peak in the
reflection plane (1 0 0) in all variations of Zn concentration. This shows that the crystal structure
obtained is spherical.
[13] carried out SEM testing on samples with several variations of zinc acetate added to
lime extract (Citrus aurantifolia). The non-uniform surface morphology was clearly visible from
the lowest concentration (0.05 M), but it also showed that no particles were formed. When the
concentration of Zn in the orange extract was increased to 0.10 M, it could be seen that the
particles agglomerated at certain places and the diameter of the particles varied roughly between
0.15 m and 0.35𝞵m. [10] also conducted tests using SEM to determine the morphology of the
material. From the SEM results, it can be seen that ZnO is in the form of nanoparticles with sizes
ranging from 17 nm - 34 nm. [12] showed SEM results with a ZnO nanoparticle size of about 17
± 5 nm and exhibited an excellent nearly spherical shape with a narrow size distribution. In
addition, the resulting nanoparticles are much smaller than other researchers.
4.2 Electrocatalytic Properties of Glucose Biosensors

techniques were used cyclic voltammetry and amperometry to determine the

electrocatalytic oxidation activity of glucose. In research [10], the GOx-ZnO/CPE biosensor was
tested using 2 types of electrodes, namely CPE (carbon paste electrode) and CPE modified by
ZnO NPs. A buffer solution of 0.1 M PBS (pH 7) was dripped onto the electrode with added 10
mM H2O2 which was oxidized.test results showed an increase in current response with H2O2 at
both electrodes, but a significantly larger oxidation peak was observed for ZnO/CPE.
Amperometric response of both electrodes (CPE and ZnO/CPE) also increased with the addition
of H2O2 from 0.5 mM to 10 mM.H2O2 almost fourfold higherTherefore, the synthesized green
ZnO nanoparticles can catalyze and accelerate the H2O2 oxidationandservethus as an excellent
glucose biosensor. In addition to evaluating the test with variations in the addition of H2O2 , the
effect of increasing the number of ZnO NPs into CPE for the manufacture of glucose biosensors
was also investigated.response amperometric the biosensor with 1.0 mM glucose increased with
increasing the number of nanoparticles from 2-4 mg, whereas a further increase in the number of
nanoparticles led to a decrease in the amperometric. So that the addition of ZnO NPs as much as
4 mg is more optimal to be carried out in other experiments.

[11] also optimized the electrocatalytic activity of the developed

ZnO/PIn-MWCNTs/Frt/GOx bioanodes by varying the glucose concentration from

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 6

10-60mM.activity catalytic to glucose concentration was carried out using a linear sweep
voltammetry (LSV) technique. From the test results, there was a gradual increase in the height of
the oxidation peak as the substrate concentration (glucose) increased up to 50 mM. These results
indicate that the synthesized ZnO/PIn-MWCNTs nanocomposite has sufficient binding sites due
to its porous network structure which enhances its enzyme loading and catalytic activity.

The decrease in the observed peak indicates the saturation kinetics of the reaction. This
confirmed that the filling of all voids of the nano biocatalyst was not affected by a further
increase in concentration. Moreover, the optimized current density obtained via glucose
bioelectrolysis is 4.9 mA cm-2 at a scanning speed of 100mVs-1 by consuming 50 mM glucose in
PBS pH 7.4 as can be seen in the calibration curve b plotted against anodic current versus
concentration glucose.

In addition to the effect of the applied voltage and glucose concentration parameters,
temperature also affects the sensitivity of the glucose biosensor. In study [12] the effect of
varying temperature on the response of the GCE/ZnO DCS/GOx biosensor was also examined
between 23 and 65°C. The current response gradually increases with increasing temperature and
reaches its maximum value at 37 °C. This is because the enzyme activity increases at higher
temperatures. After 40 °C, the response decreases, which is due to the natural thermal
degradation of GOx. Although the GCE/ZnONDCS /GOx biosensor showed a good response at
37 °C, room temperature was still chosen for this work to prevent possible escape of evaporation
at higher temperatures and ease of processing.

5. Conclusion
From the several methods of synthesizing ZnO nanoparticles above, it can be concluded
that the most effective methods to use are MPC (Mechano Chemical Process) and PVD (Physical
Vapor Deposition) because the process is quite simple and costs low. In addition, the length of
milling time in the ball milling and the temperature used in the vapor can affect the size of the
resulting nanoparticles.
The green synthesis is effective in the formation of ZnO nanoparticles and is more
environmentally friendly when compared to physical and chemical synthesis methods.
Coprecipitation methods and hydrothermal methods are generally used in green synthesis. In
several studies, it was found that ZnO NPs were found in samples which were indicated by the
presence of a peak around 360 nm on UV-vis testing. In addition, XRD and SEM tests showed a
high hexagonal crystal structure on ZnO NPs with crystal sizes ranging from 17 - 34 nm.
However, in several studies, ZnO NPs have a larger size of about 100 nm and have a spherical
On the other hand, glucose biosensors prepared from several studies have shown good
electro-catalytic ability for glucose determination, in addition to using the green synthesis ZnO
NPs can increase the sensitivity of the biosensor.

6. Acknowledgment
Praise God Almighty, for the plenty presence of mercy and his grace, so that the writers can
complete the journal review with the title : “Green Synthesis of ZnO Nanoparticles for Glucose
Biosensor Applications”. Appreciation and thanks the author gave to Mr. Septia Eka Marsha

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 7

Putra, S.Si, M.Eng, Ph.D and Mr. Dr. Eng Lukman Nulhakim, ST, M.Si as the lecturer of
Material Engineering subject who has guided us in writing this review.

As well as a thank you to the entire members of the group who were always helpful in drafting of
the journal review.

At the end of the word, the authors realized that there are still so many mistakes in this writing.
The journal review is not worth enough from perfection. Therefore, we invoke suggestions ans
criticisms which can build for the sake of perfection and may be useful for all of us.

7. Reference
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their biological synthesis, antimicrobial activity, uptake, translocation and
biotransformation in plants. Journal of Materials Science, 185–201. Retrieved 2022,
from https://doi.org/10.1007/s10853-017-1544-1
[2] Bayrami, A., Haghgooie, S., Pouran, SR, Arvanag, FM, & Yangjae, AH (nd). Synergistic
antidiabetic activity of ZnO nanoparticles encompassed by Urtica dioica extract. 31(5),
2110-2118. https://doi.org/10.1016/j.apt.2020.03.004
[3] Fakhari, S., Jamzad, M., & Fard, HK (2019, Jan 11). Green synthesis of zinc oxide
nanoparticles, 12. No.1. https://doi.org/10.1080/17518253.2018.1547925
[4] Hieu, NV (2017). Nanomaterials: a guide to fabrication and applications. Advances in
Natural Sciences: Nanoscience and Nanotechnology, 8(2). Retrieved 3 5, 2022, from
https://iopscience.iop.org/article/10.1088/2043-6254/aa71eb
[5] Jamdagni, P., Khatri, P., & Rana, J. (2016). Green synthesis of zinc oxide nanoparticles
using flower extract of Nyctanthes arbor-tristis and their antifungal activity, 1-8. ,
http://dx.doi.org/10.1016/j.jksus.2016.10.002
[6] Kairyte, K., Kadys, A., & Luksiene, Z. (2013). Antibacterial and antifungal activity of
photoactivated ZnO nanoparticles in suspension. Journal of Photochemistry and
Photobiology B: Biology, 128, 78-84. Retrieved 2022, from
https://doi.org/10.1016/j.jphotobiol.2013.07.017
[7] Khan, I., Saeed, K., & Khan, I. (2019, November). nanoparticles. Nanoparticles:
Properties, applications and toxicities, 12(7), 908-931. 10.1016/j.arabjc.2017.05.011
[8] Perez, PJ, Ferreria, N. d. F., dos Reis, JS, & Cruz, RS (2012). Food Bioprocess Technol.
Zinc Oxide Nanoparticles: Synthesis, Antimicrobial Activity and Food Packaging
Applications, 5, 1447-1464. 10.1007/s11947-012-0797-6
[9] Rajeshkumar, S., Venkat Kumar, S., Ramaiah, A., Agarwal, H., Lakshmi, T., & Roopa, SM
(October 2018). Biosynthesis of zinc oxide nanoparticles using Mangifera indica leaves
and evaluation of their antioxidant and cytotoxic properties in lung cancer (A549) cells.
Enzyme and Microbial Technology, 117, Pages 91-95. Retrieved 2022, from
https://doi.org/10.1016/j.enzmictec.2018.06.009
[10] DÖNMEZ, S. (2020). Green Synthesis of Zinc Oxide Nanoparticles Using Zingiber
Officinale Root Extract and Their Applications in Glucose Biosensor. El-Cezerî Journal
of Science and Engineering, 7, 1191 - 1200.https://doi.org/10.31202/ecjse.729462
[11] Inamuddin, Shakeel, N., Ahamed, MI, Kanchi, S., & Kashmery, HA (2020). Green

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 8

 synthesis of ZnO nanoparticles decorated on polyindole functionalized-MCNTs and
used as anode material for enzymatic biofuel cell applications. Scientific Reports |, 1 -
10. https://doi.org/10.1038/s41598-020-61831-4
[12] Mutuchamy, N., Atchudan, R., Nesakumar, T., Perumal, S., & Lee, YR ( 2018).
High-performance glucose biosensor based on green synthesized zinc oxide
nanoparticle embedded nitrogen-doped carbon sheet. Journal of Electroanalytical
Chemistry, 195 - 204. https://doi.org/10.1016/j.jelechem.2018.03.059
[13] Samat, NA, & ​Nor, RM (nd). Sol–gel synthesis of zinc oxide nanoparticles using Citrus
aurantifolia extracts,. Sol–gel synthesis of zinc oxide nanoparticles using Citrus
aurantifolia extracts,, 39(ISSN 0272-8842), 545-548.
https://doi.org/10.1016/j.ceramint.2012.10.132.

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 9

8. Appendix
8.1 Summary 1 : Nanang Faddilah

Journal Authors : Inamuddin, Nimra Shakeel, Mohd ImranAhamed, Suvardhan Kanchi ,

HebaAbbas Kashmery
Tittle : Green synthesis of ZnO nanoparticles decorated on polyindole
functionalized-MCNTs and used as anode material for enzymatic
biofuel cell applications
Journal Name : Scientific Reports
Volume : 10
Page : 1 - 10
Year : 2020

Currently, one of the most important aspects for the development of enzymatic biofuel
cells (EBFC) is the synthesis of new electrode materials which have high current density,
low open circuit voltage (OCV), and long stability. . To achieve some of the parameters
above, many new strategies are used by researchers for the development of advanced
materials. Currently, nanomaterials and nanocomposites are promising materials that
have been used as effective electrode materials in the application of solar cells,
supercapacitors and biofuel cells. Here, we account for a novel electrocatalyst as an
electrode material consisting of ZnO nanoparticles decorated on a polyindole
(PIn)-multi-walled carbon nanotube (MWCNT) surface, for enzyme immobilization and
a glucose oxide (GOx) mediator (Ferritin).

Synthesis of ZnO NPs using neem leaf extract. Te Neem (A. indica) leaves were
collected and washed with tap water to remove dirt and other contaminated organic
matter, followed by distilled water and air dried at room temperature. The finely chopped
leaves were stored in a beaker containing double distilled water and boiled for 30
minutes. The extract was cooled and filtered through Whatman filter paper no. 1, and the
extract was stored at 4 °C after being covered with aluminum foil for further use. First,
10 mL of neem extract was mixed with 90 mL of 0.1 M zinc nitrate solution dropwise
under continuous stirring at room temperature for 4-5 hours. After that, 50 mL of 2.0M
NaOH solution was added dropwise to the above mixture and allowed to continue
stirring for 2 hours. The resulting white precipitate was filtered and washed repeatedly
with distilled water followed by ethanol to remove impurities. Finally, a white powder
was obtained after overnight drying of the pure precipitate at 60 °C in an oven. Dried
ZnO Nps were stored for further analysis for the manufacture of nanocomposites
(ZnO/PIn-MWCNTs).

-MWCNT fabrication process was prepared by chemical oxidative in situ polymerization

of indole on the surface of the MWCNT and assessed by various techniques. From the
results of Scanning Electron Microscopy (SEM) shows the morphology of MWCNT that
are interconnected in the polymer matrix. XRD and FTIR assays, confirmed the
crystallinity and distinct functional groups available in the synthesized materials,
respectively. Electrochemical assessment showed that the ZnO/PIn-MWCNT/Frt/GOx
nano bio-catalyst showed significantly higher electrocatalytic activity against glucose

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 10

 oxidation with a maximum current density of 4.9 mA cm−2 with a glucose concentration
of 50mM in phosphate buffer solution (PBS) (pH). 7.4) as a test solution by applying a
scan speed of 100 mVs−1. The results reflect that the prepared ZnO/PIn
MWCNTs/Frt/GOx biocomposite is a promising bioanode for the development of EBFC

8.2 Summary 2: Nanang Faddilah

Journal Authors : Nallal Muthuchamya, Raji Atchudanb, Thomas Nesakumar Jebakumar

Immanuel Edisonb, Suguna Perumalc, Yong Rok Lee
Tittle : High-performance glucose biosensor based on green synthesized zinc oxide
nanoparticle embedded nitrogen-doped carbon sheet
Journal Name : Journal of Electroanalytical Chemistry
Volume : 816
Page : 195–204
Year : 2018

The enzymatic glucose sensor has recently been manufactured with a highly selective,
sensitive and stable on glass carbon electrodes (GCE) using zinc oxide (ZnO) nanoparticles
embedded nitrogen-doped carbon sheets(ZnONDCS). In the fabrication process, first ZnO
NPs were synthesized by a simple hydrothermal method.synthesis procedure ZnO ,
approximately 200 mg of ZnP was added to 30 mL of peach, followed by 1 mL of aqueous
ammonia in a Teflon Lined stainless steel autoclave with a volume capacity of 50 mL.
Then, the autoclave was tightly closed and maintained at 180 °C for 12 hours. After
hydrothermal treatment, the autoclave was naturally cooled to room temperature. The
resulting product was filtered and washed with DI water several times, then the residue
was dried at 70 °C overnight. Completely dry powder samples were stored and used for
further analysis.

The physicochemical bonding of the synthesized ZnONDCS nanocomposite was

investigated by XRD, Raman, ATR-FTIR, FESEM, and HRTEM techniques. The N-atoms
that were effectively bonded to the graphite carbon structure with an atomic percentage of
7.5% were analyzed by ATR-FTIR and XPS. In addition, the well-distributed ZnO NPs on
carbon sheet were clearly seen from HRTEM and FESEM by elemental mapping. In
addition, glucose oxidation (GOx) was successfully immobilized on ZnO@NDCS
nanocomposite. The developed GCE/ZnO@NDCS/GOx showed excellent electrooxidation
performance against glucose at pH 7.0 PBS. In addition, the fabricated
GCE/ZnO@NDCS/GOx biosensor showed good analytical features such as high
sensitivity (231.7 A mM−1 cm−2), very low detection limit (6.3 M), wide linear range (0
,2–12 mM), and amperometric (3 s) along with good selectivity, repeatability,
reproducibilityand long-term storage stability. In addition, the sensor made also shows
good selectivity for glucose with the addition of the concentration of the

GCE/ZnONDCS/GOx Biosensor, which is quite stable, selective, and successfully applied

to quantitative monitoring of glucose in human blood serum. The synthesized ZnONDCS
nanocomposites could be useful in other applications in the field of solar cells and
optoelectronic devices. The results of this study indicate a simple and effective method of

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 11

 obtaining electrode materials for enzymatic glucose sensors.

8.3 Summary 3 : Nanang Faddilah

Journal Authors : Soner

DÖNMEZ : Green Synthesis of Zinc Oxide Nanoparticles Using Zingiber
Officinale Root Extract and Their Applications in Glucose Biosensor

Journal Name : El-Cezerî Journal of Science and Engineering

Volume : Vol: 7, No: 3
Page : 1191-1200
Year : 2020

Green Synthesis of nanoparticles through plant extracts has become an important research
area in nanotechnology. In this study, a novel amperometric glucose biosensor based on
zinc oxide (ZnO) nanoparticles synthesized using ginger root (Zingiber officinale) was
created. In the synthesis process Zn(CH3COO)2.2H2O was dissolved in 50 mL of distilled
water to produce a final concentration of 0.02 M. Then, 20 mL of Z. officinale root extract
(0.25 g/mL in distilled water) was slowly added. into a solution of Zn(CH3COO)2.2H2O.
This mixture was heated at 80 °C under continuous stirring with a magnetic stirrer for
about 2 hours, until a white precipitate was formed. These ZnO nanoparticle precipitates
were centrifuged for 10 min at 10,000 rpm, washed with distilled water, and then washed
with methanol to remove unwanted impurities. The ZnO nanoparticles were dried (at 60 °C
overnight) and then calcined at 450 °C in a furnace for 2 h. Glucose oxide (GOx) was
immobilized onto a ZnO-modified carbon paste (CPE) electrode via cross-linking with
glutaraldehyde. The prepared biosensor (GOx-ZnO/CPE) showed good electrocatalytic
capability for glucose determination. The biosensor also exhibited a low detection limit
(14.7 M), fast response (less than 1 s), high sensitivity (15.98 A mM−1 cm−2), and higher
biological affinity (Michaelis−Menten constant estimated to be 0 .99 mM). In addition, the
prepared biosensor showed good anti-interference ability in relation to ascorbic acid (AA)
and uric acid (UA). These results indicate that a simple and cost-effective biosensor has
been developed for the determination of glucose

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 12

8.4 Summary 4: Salsabila Regina Mayanti
Journal authors : Paula Judith Perez Espitia, Nilda de Fátima Ferreira Soares, Jane Sélia
d dos Reis Coimbra, Renato Souza Cruz, Nélio José de Andrade, Eber A
Antonio Alves Medeiros
Tittle : Zinc Oxide Nanoparticles: Synthesis, Antimicrobial Activity and Food
P Packaging Applications
Journal Name : Food Bioprocess Technol
Volume :5
Page : 1447-1464
Year : 2021

Based on the increasing problems regarding global public health in 2011 caused by
foodborne disease, a food packaging with nano technology was created that can
maintain the quality of the food in it in order to control foodborne pathogens. This
packaging is commonly referred to as antimicrobial packaging. Serves to reduce and
inhibit the growth of microorganisms on the surface of food.

The synthesis of ZnO nanoparticles was carried out using two main methods, namely
MCP (Mechano Chemical Process) and PVD (Physical Vapor Deposition). In the MCP
method, ZnCl2 and Na2CO3 to produce ZnCO3 and NaCl.

Reaction: ZnCl2 + Na2CO3 + 8 NaCl → ZnCO3 + 10NaCl

The mixture is heated at a temperature of 170-380𝑡𝑜 decompose ZnCO3 into ZnO. The
resulting nanoparticles are 20-30 nm in size. The size of the resulting nanoparticles
depends on the grinding time and the temperature used. Optimal conditions were
achieved by using a milling time of 40 minutes.

In the PVD method, reactant gases are added to the vapor and cooled at a controlled
rate and condensed to form nanoparticles. The resulting nanoparticles are very dense
and discrete and have a size of 8-75 nm.

This study concluded that ZnO nanoparticles can be effective antimicrobial agents due
to their activity against various microorganisms and high resistance to complex
processing conditions. In addition, ZnO nanoparticles can retain their antimicrobial
activity even when incorporated in polymer matrices indicating their potential for food
preservation through their use as antimicrobial packaging. Besides being antimicrobial,
ZnO has presented modifications in the structure and properties of packaging materials
such as mechanical and thermal resistance. This type of characterization is relevant in
food packaging design. Because each characteristic can affect the physical quality of
the developed packaging.

However, antimicrobial food packaging developed using nanotechnology can also raise
new concerns for society. Several studies are currently investigating the ability of ZnO
nanoparticles in antimicrobial packaging to transfer to food. So that the toxicological
impact of ZnO nanoparticles must also be evaluated to determine its positive and

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 13

 negative effects on food safety.

8.5 Summary 5: Salsabila Regina Mayanti

Journal authors : Pragati Jamdagni, Poonam Khatri, JS Rana
Tittle : Green synthesis of zinc oxide nanoparticles using flower extract of N
Nyctanthes arbor-tristis and their antifungal activity
Journal Name : Journal of King Saud University – Science
Volume :2
Page : 1-8
Year : 2016

Biological reducing agents are being explored worldwide to minimize the toxic effects
of chemicals used in nanoparticle fabrication. This study states a green approach for the
synthesis of ZnO nanoparticles using Nyctanthes arbor-tristis. Flower extract was used
as a biological reducing agent to synthesize ZnO nanoparticles from Zn(CH3COO)2.
nanopowder was characterized using various analytical techniques, such as UV–Visible,
FTIR, XRD, and TEM.

Synthesis was carried out by mixing 0.01M Zn(2CH3COO)and flower extract. pH was
maintained at 12 while continuously stirring for 2 hours. This results in a white
precipitate which is then dried at 60℃ overnight. Prior to drying, the precipitate was
first centrifuged at 15000 rpm for 5 minutes and washed twice using DI water. The
concentration of Zn(CH3COO)2 from 0.025 M to 0.2 M and the volume of flower
extract from 0.25 mL to 2 m. The solution was continuously stirred using a magnetic
stirrer and maintained at an increase in pH of 9, 10, 11, 12, and 13 using a 2 M NaOH
solution and the reaction temperature was maintained at 60, 70, 80, and 90℃.

Increasing the concentration of 0.02M resulted in a decrease in absorbance and a

substantial expansion of the peak. So it can be seen that increasing the concentration of
metal ions beyond the threshold value causes a decrease in the synthesis of
nanoparticles.

The nanoparticle size range obtained on synthesis under optimal conditions was 12-32
nm as reported by TEM and also showed that the nanoparticles were present in the form
of aggregates. In addition to the morphological analysis, the TEM analysis also
established the role of the lower-intensity overburden on the NP surface. XRD studies
confirmed the crystalline nature of the nanoparticles showing particle sizes in the range
provided by electron microscopy data. The nanoparticles were tested for antifungal
potential and found to be active against all five phytopathogens tested with the MIC
Value recorded as 16 lg/mL. It can be exploited to develop antifungal agents for
commercial use in agriculture. This research proves an environmentally friendly
approach for the synthesis of ZnO nanoparticles.

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 14

8.6 Summary 6 : Reni Silvia Dewi
Journal Authors : Soner DÖNMEZ
Title : Green Synthesis of Zinc Oxide Nanoparticles Using Zingiber
Officinale Root Extract and Their Applications in Glucose Biosensor
Journal Name : El-Cezerî Journal of Science and Engineering
Volume : Vol: 7, No: 3
Page : 1191-1200
Year : 2020

Various applications of glucose biosensors have been prepared with chemically

synthesized ZnO nanoparticles, only limited investigations have focused on glucose
biosensors prepared with green synthesized ZnO nanoparticles. This journal explains,
green synthesis of ZnO nanoparticles using Zingiber officinale root extract and the use
of these nanoparticles to make glucose biosensors. In this study, an amperometric
glucose biosensor based on green synthesized zinc oxide (ZnO) nanoparticles using
Zingiber officinale roots. Glucose oxidase (GOx) was immobilized to a ZnO-modified
carbon paste (CPE) electrode via cross-linking with glutaraldehyde. All electrochemical
experiments in this study were carried out using the CHI 1230B electrochemical
working method. The three-electrode system consisted of an Ag/AgCl reference
electrode, a ZnO nanoparticle modified carbon paste electrode as a working electrode,
and a platinum electrode as an auxiliary electrode. The experimental results show that
the synthetic green ZnO nanoparticle modified glucose biosensor performs much better
than many previously published ZnO nanoparticle based biosensors in terms of
sensitivity, response time, detection limit and cost effectiveness. The prepared
biosensor (GOx-ZnO/CPE) showed good electrocatalytic capability for glucose
determination. The biosensor also exhibited a low detection limit (14.7 M), fast
response (less than 1 second), high sensitivity (15.98 A mM).1cm2), and higher
biological affinity (Michaelis−Menten constant was estimated at 0 99 mM). In addition,
the prepared biosensor showed good anti-interference ability in relation to ascorbic acid
(AA) and uric acid (UA). These results indicate that glucose determination has been
successfully made from a simple and cost-effective biosensor.

8.7 Summary 7 : Reni Silvia Dewi

Journal authors : NurulAin Samat and RoslanMd Nor
Tittle : Sol–gel synthesis of zinc oxide nanoparticles using Citrus aurantifolia extracts
Journal Name : Ceramics International
Volume : 39
Page : 545-548
Year : 2013

In this journal we report the synthesis of ZnO NPs have been successfully
synthesized using Citrus aurantifolia extract at different concentrations of Zn acetate
through the sol-gel method. Citrus aurantifolia fruit that has been peeled, cut into small
pieces and mixed with deionized water. Puree until it becomes mush, then filtered. Use
a muslin cloth to remove any solid particles. After becoming a liquid, then filtered
using a filter syringe with a pore size of 200 nm. Zinc acetate is dissolved in 100 ml of

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 15

 solution. The liquid extract of C. aurantifolia was filtered based on the concentrations
of 0.05 M, 0.10 M, 0.15 M and 0.20 M. Its pH value was between 3.7 and 4.0. The
mixture is then heated to 90° C. with continuous stirring for 3 hours until a white
precipitate appears. The precipitate was washed with deionized water and dried at
100°C for 6 hours. XRD analysis of the XRD pattern at 0.10 M showed a similar XRD
pattern to 0.15 M and 0.20 M. For all samples, the peaks of the reflection (1 0 0) plane
were dominated by peaks because (1 0 1), ( 1 0 2) and (1 1 0) seem calm. It seems that
the ZnO NPs have a preferential orientation at (1 0 0).
From the SEM images for ZnO synthesized at 0.15 M and 0.20 M it looks
uniform with a structure around 100 nm. In general, we have shown that C. aurantifolia
extract is feasible for the synthesis of ZnO NPs utilizing a sol-gel technique in which
such biosynthesis offers an alternative to green synthesis methods.

8.8 Summary 8 : Pipit Fitria Yuliadi

Journal authors : Ibrahim Khan, Khaleed Saeed and Idrees Khan
Tittle : Nanoparticles: Properties, applications and toxicities
Journal Name : Arabian Journal of Chemistry
Volume : 12
Page : 908 - 931
Year : 2019

This journal provides details about types, synthesis, characterization,

physiochemical and applications of nanoparticles in various fields. Nanoparticles are
small materials that range in size from 1 to 100 nm. Nanoparticles can be classified into
different types based on their nature, shape, or size. Nanoparticles are divided into
several different groups including fullerenes, metal nanoparticles, ceramic
nanoparticles, and polymer nanoparticles. Nanoparticles have unique physical and
chemical properties due to their high surface area and nanosize. Reactivity, and other
properties also depend on its unique size, shape and structure. Due to these
characteristics, nanoparticles are suitable materials for application in a wide variety of
fields, including catalysis, medical applications, energy-based research, and
environmental applications. Through different characterization techniques such as
SEM, TEM and XRD, it was revealed that the nanoparticles have a size range from a
few nanometers to 500 nm. While the morphology can also be controlled. Due to their
small size, nanoparticles have a large surface area, which makes them suitable
candidates for various applications. Synthetic techniques can be useful for controlling
certain morphology, size and magnetic properties of nanoparticles.
Although nanoparticles are useful for many applications, there are still some
health hazards due to their uncontrolled use and disposal into the natural environment,
which should be considered to make the use of nanoparticles more convenient and
environmentally friendly. Suggestions for future researchers are to carry out reactions
with different parameters such as temperature, pressure, time, and pH to control the
shape and morphology of the resulting nanoparticles, so that various parameters must
be optimized to achieve certain product characteristics. In addition to good
implications, learn certain characterization techniques, and should be careful when
using materials such as heavy metals that are susceptible to environmental hazards and

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 16

 can also affect life.

8.9 Summary 9 : Pipit Fitria Yuliadi

Journal authors : Shabnam Fakhari, Mina Jamzad and Hassan Kabiri Fard
Tittle : Green synthesis of zinc oxide nanoparticles
Journal Name : Green Chemistry Letters and Reviews
Volume : Vol.12, No.1
Page : 19-24
Year : 2019

Green synthesis of nanoparticles by biological systems especially plant extracts

has become a new field in nanotechnology. ZnO nanoparticles were synthesized using
Laurus nobilis L.salts zinc (zinc acetate and zinc nitrate) as precursors. The synthesized
nanoparticles were characterized by Ultraviolet–Visible spectroscopy (UV–Vis),
Fourier Transform Infrared Spectroscopy (FT-IR), X-Ray Diffraction (XRD) analysis,
Energy-Dispersive X-ray analysis (EDX) and Scanning Electron Microscopy (SEM). ).
The UV–Vis spectra show typical absorption peaks around 350 nm due to their high
excitation bond energy at room temperature.
The chemical bond formation of zinc oxide was confirmed by FTIR analysis.
The XRD results revealed the formation of a hexagonal wurtzite structure, and SEM
analysis showed a spherical shape with an average size (21.49, 25.26) nm for the
nanoparticles synthesized by zinc acetate and zinc nitrate respectively. EDX analysis
confirmed the high purity for the synthesized nanoparticles. The EDX results confirmed
the presence of zinc and oxygen in the synthesized ZnO nanoparticles. The FTIR study
demonstrated the formation of ZnO and showed that the plant extracts contained
various phytochemicals, which worked as capping and stabilizing agents for the
synthesized ZNPs. From the analysis of the results, it is clear that the precursor has
played an important role in the morphology and surface structure of the ZnO
nanoparticles. Our results confirm the potency of nobilis L.L.for the synthesis of ZnO
nanoparticles in a simple, fast and environmentally friendly way.

Green Synthesis of ZnO Nanoparticles for Glucose Biosensor Applications. 17