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Recent advances in electrochemical non-enzymatic glucose sensor for the

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DOI: 10.1016/j.sna.2023.114778

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 Sensors & Actuators: A. Physical 363 (2023) 114778

Contents lists available at ScienceDirect

Sensors and Actuators: A. Physical

journal homepage: www.journals.elsevier.com/sensors-and-actuators-a-physical

Recent advances in electrochemical non-enzymatic glucose sensor for the

detection of glucose in tears and saliva: A Review
Kermue Vasco Jarnda a, Danqi Wang a, Qurrat-Ul-Ain a, Richmond Anaman b,
Varney Edwin Johnson b, Garmai Prosperity Roberts c, Pauline Sammumah Johnson d,
Bob Wisdom Jallawide Jr b, Tianhan Kai a, *, Ping Ding a, *
a
Department of Health Inspection and Quarantine, XiangYa School of Public Health, Central South University, Changsha, Hunan Province 410078, People’s Republic of
China
b
School of Metallurgy and Environment, Central South University, Changsha, Hunan Province 410083, People’s Republic of China
c
Department of Nursing, XiangYa School of Nursing, Central South University, Changsha, Hunan Province 410078, People’s Republic of China
d
Department of Social Medicine and Health Management, XiangYa School of Public Health, Central South University, Changsha, Hunan Province 410078, People’s
Republic of China

A R T I C L E I N F O A B S T R A C T

Keywords: Biosensors have been extensively investigated and used in a wide range of applications, from environmental
Detection monitoring to clinical diagnostics. Glucose biosensors have gained popularity and application because glucose
Non-enzymatic sensing is critical for diabetes diagnosis. However, traditional methods for diabetes monitoring are unpleasant,
Electrochemical sensing
expensive, and invasive. To address this issue, non-enzymatic glucose sensors have been developed for tear and
Glucose biosensors
saliva-based detection. In this review, we comprehensively cover the recent advances in these sensors and
Saliva
Tears explain the principles and mechanisms of electrochemical glucose detection. We also talk about the recent
development of metal alloy-based materials that work well as electrochemical glucose sensors. These materials
have pore channels that can be changed, a large specific surface area, and metallic active sites that are spread
out. Furthermore, we discuss the promising potential of metal alloy-based materials as efficient electrochemical
glucose sensors. These materials have tuneable pore channels, a large specific surface area, and well-dispersed
metallic active sites, which can enhance the sensitivity and selectivity of glucose detection.

1. Introduction
The medical application of blood glucose sensing has drawn
increasing attention to glucose sensors. The amount of blood glucose
must be carefully monitored and controlled, especially for diabetes in­
dividuals. [182]. In 2014, it was estimated that 422 million people
globally will have diabetes; by 2040, that number might rise to 642
million. [142]. Therefore, developing a quick and accurate way to
measure glucose levels would be significant [105]. After cancer and
cardiovascular illnesses, diabetes has emerged as the third chronic,
non-communicable disease to pose a threat to human health. [42].
Physiological blood glucose levels are between 70 and 120 mg/dL [30],

Abbreviations: Ag, Silver; Au, Gold; Aunps, Gold Nanoparticles; C-1, Carbon molecule; C-1, Carbon One; Celif, Capillary Electrophoresis-Laser-Induced Fluores­
cence; CI, Chloride ions; Cnts, Carbon Nanotubes; COFs, Covalent Organic Framework; CoO, Cobalt Oxide; CoP, Cobalt Phosphide; CoS, Cobalt Sulfide; Cu, Copper;
CuO, Copper Oxide; DES, Dry Eye Syndrome; DFT, Density functional theory; DM, Diabetes Mellitus; EIS, Electrochemical Impedance Spectroscopy; FAD, Flavin
Adenine Dinucleotide; Fe, Ferric Oxide; Fe3O4, Magnetite; G, Glucose; Gce, Glassy Carbon Electrode; GDH, glucose dehydrogenase; Gox, Glucose Oxidase Enzyme;
GSH, Glutathione; H2O, Water; H2O2, Hydrogen Peroxide; H2Q, Hydroquinone; HPLC-Pad, High-Performance Liquid Chromatography Combined with Pulsed
Amperometric Detection; IHOAM, Incipient Hydrous Oxide Adatom Mediator; Lod, Limit of Detection; LC-ESI-MS, Liquid chromatography-electrospray ionization-
tandem mass spectrometry; MHz, Megahertz; Mn, Manganese; Mn, Manganese Oxide-; MOSFET, Metal Oxide Silicon Field Effect Transistors; MPA, Mercaptopro­
pionic Acid; MWCNTs, Multi-Walled Carbon Nanotubes; NaOH, Sodium Hydroxide; NEGS, Non-Enzymatic Glucose Sensor; Ni, Nickel; Oh, Hydroxyl Ion; Pani,
Polyaniline; PBQ, Parabenzoquinone; PBS, Phosphate Buffer Solution; Pd, Palladium; pKa, logarithmic dissociation constant; Pt, Platinum; Rct, Charge Transfer
Resistance; rGO, graphene oxide; Rh, Rhodium Oxide-Based; SPR, Surface Plasmon Resonance; SAM, Self-Assembled Monolayer; Sio2, Silicon Dioxide; Swcnts, Single-
Walled Carbon Nanotubes; Ti, Titanium Oxide; TMP, Tetramethylpyrazine; ZnO, Zinc Oxide.
* Corresponding authors.
E-mail addresses: th_kai@csu.edu.cn (T. Kai), pingding2010@163.com (P. Ding).

https://doi.org/10.1016/j.sna.2023.114778
Received 4 July 2023; Received in revised form 22 October 2023; Accepted 25 October 2023
Available online 30 October 2023
0924-4247/© 2023 Published by Elsevier B.V.
K.V. Jarnda et al.
whereas salivary physiological glucose concentrations range from 0.05
to 21.61 mg/dL [43]. Large fluctuations in G levels can result in a
number of secondary problems, including injury to several organs,
including the heart and brain [59], the kidneys [6,22], and the nerves
[37]. Measuring the amount of glucose in the blood is important for
preventing and treating clinical diabetes, since too much glucose in the
blood is the main cause of diabetes [181].
Numerous techniques, including the fluorescence method [50],
colorimetry [220], and electrochemistry [7,118], have been developed
to quantify blood glucose concentration. It is necessary to puncture the
skin or a peripheral capillary to take samples, which could hurt the
patients. Small blood samples are typically taken using a finger-stick
instrument, and then H202, which is created by the enzymatic oxida­
tion of glucose, is indirectly detected electrochemically. Patients’
discomfort and agony are caused by inadequate glucose regulation,
which breeds subsequent diseases [61]. Finger-prick tests can disrupt
continuous measurement and result in hypo- and hyperglycemia [188].
Therefore, quick, painless, and frequent blood glucose measurement
techniques would be of significant interest to both individuals and
medical and societal communities. A number of initiatives have made it
possible to create non-invasive techniques for identifying certain
chemical biomarkers in human fluids, such as sweat, tears, urine, or
saliva [178]. Monitoring saliva and tears is becoming more and more
important as a non-invasive way to find more diseases. The most
interesting thing about saliva is that it can show signs of disease,
including those that are often found in the blood [113].
Hoa Duc Trinh et al. created CS@SiO2 core-shell nanoparticles with
GOx to detect salivary glucose [113]. However, the presence of con­
taminants significantly impacted the accuracy of the test results.
Attention has been drawn to tear fluid as a non-invasive bodily fluid. In
addition to being non-invasive, the tear environment is cleaner than the
blood environment, and tear samples don’t need to be treated before­
hand, making the procedure straightforward. Blood glucose levels and
tears’ glucose levels are highly positively associated [159]. Thus, tear
fluid glucose measurement is non-invasive. Tear glucose has been
detected in several ways for decades. Capillary
electrophoresis-laser-induced fluorescence (CELIF) [194], HPLC-PAD
[137], and liquid chromatography-electrospray ionization-tandem
mass spectrometry (LC-ESI-MS) are examples [85].
Zhang et al. [243] developed a novel glucose detection method using
citric acid-coated silver nanoparticles. GOx may oxidize the samples’
glucose to gluconic acid and H2O2, according to their method⋅H2O2
oxidizes silver nanoparticles to release Ag+ and citric acid, increasing
the solution’s ionic strength. Although the chemiluminescence assay has
improved and these electrochemical sensors have an appealing design,
their low sensitivity prevents their practical usage. However, the pre­
ceding approaches need nanoparticles, which are expensive and
time-consuming to manufacture. The production conditions of these
nanoparticles are prone to change, which over time complicates
research.
Diabetes management and treatment require regular glucose moni­
toring with sensors [165,218]. Enzymatic and non-enzymatic glucose
sensors are already available [110,138,166,215,44]. Non- enzymatic
glucose sensors are quick, reliable, and sensitive [213,215]. Due to their
limitations, non-enzymatic glucose sensors are low-specific. Tempera­
ture, pH, and other environmental variables affect glucose oxidase
[240]. Adsorption, cross-linking, trapping, and electro-polymerization
make enzyme immobilization on electrode substrates difficult [217].
The importance of non-enzymatic glucose sensors cannot be overstated;
however, Pd and Ag are components of non-enzymatic glucose sensors
[80]. A simple electrochemical method [227,242,71] and
non-enzymatic glucose electrodes can also measure glucose concentra­
tion in human physiological fluids. This review mainly discusses dia­
betes and blood non-enzymatic glucose monitoring in tears and saliva,
electrochemical principles of glucose sensors, including nonenzymatic
glucose detection methods, recent prospects in non-enzymatic glucose
2
Sensors and Actuators: A. Physical 363 (2023) 114778
sensors, metal alloy-based glucose sensors, such as COFs, MOFs, and
other materials, as well as opportunities for their further development
and applications.
Fig. 1 shows an electrochemical glucose concentration detector. The
yellow spheres on the dark blue square electrode plate in Fig. 1a
represent the electroactive sites, which are crucial for glucose electro-
oxidation in electrochemical cells. The polygonal green target mate­
rials suggest glucose molecules in human physiological fluids like elec­
trolytes. Electroactive materials’ surfaces electro-oxidize target
materials. Second, after leaving the electrochemical cell and entering
the external circuit, the transducer and amplifier handle the electro-
oxidation reaction current (Fig. 1b). Finally, the signal processor con­
verts the recorded current into glucose concentration (Fig. 1c). This idea
led to a full evaluation of the non-enzymatic electrochemical measure­
ment of glucose in saliva and tears for use in health care and medicine.
2. Diabetes and blood non-enzymatic glucose monitoring device
Controlling diabetes requires managing its life-threatening symp­
toms. By reducing hypoglycemia and hyperglycemia as well as their
accompanying adverse effects, diabetics can live much longer [97].
Glucose monitoring can optimize patients’ treatment regimens and
provide benefits for medications, physical exercise, and nourishment
[116]. Blood glucose monitoring is the most straightforward method of
obtaining a glucose sample without being intrusive [24]. In 1962, Clark
and Lyons from Cincinnati Children’s Hospital introduced the first
glucose biosensors [48]. These sensors utilize glucose oxidase (GOx) via
electrochemistry [152]. Due to their high sensitivity (M to mM),
repeatability, and low price [213], electrochemical sensors were utilized
for blood glucose readings. GOx, the enzyme of the sensor, is
glucose-specific. Hexokinase and glucose-1-dehydrogenase, which are
uncommon enzymes, were used to detect glucose [167], but GOx is
resistant to extreme pH, temperature, and ionic strength. It must tolerate
these circumstances for production [48]. GOx converts glucose to glu­
conolactone in the presence of oxygen, producing water and H2O2 [3].
Gluconolactone and water produce carboxylic acid and gluconic acid,
respectively. To oxidize, GOx reacts with flavin adenine dinucleotide
(FAD+). The reduction of FAD+ to FADH2 through redox reactions [47]⋅
H2O2 from oxygen regenerates FAD+⋅H2O2 and glucose are determined
by electrons at the anode [135].
2.1. Tear Glucose based on non-enzymatic sensing
In Point-of-Care (PoC) diagnostics, the biochemical examination of
external bodily fluids, such as sweat, saliva, tear fluid, urine, etc., has
recently gained a lot of interest [174], as this permits non-invasive and
painless procedures [174]. Because of its high concentration of glucose,
electrolytes, proteins, and other substances compared to other external
body fluids, tear fluid offers a significant potential for non-invasive di­
agnostics [189]. Literature has documented a number of tear sensor
examples, most of which are intended to measure glucose levels enzy­
matically [163]. The creation of cutting-edge glucose sensors is essential
for biomedical research since monitoring glucose has significant medical
benefits for the treatment of diabetes [40].
When comparing the amounts of glucose in tear fluid, non-enzymatic
glucose sensors use different principles than enzymatic sensors. For
measuring glucose, these sensors commonly use electrochemical or op­
tical techniques [174]. Non-enzymatic glucose sensors in electro­
chemical sensing use certain electrode materials that react to the
presence of glucose. Metal oxides with a high affinity for glucose mol­
ecules, such as copper oxide (CuO), zinc oxide (ZnO), or tungsten oxide
(WO3), can be found in these substances [231]. A quantifiable electrical
signal is created when glucose interacts with the electrode surface to
start an electrochemical reaction [89]. Next, the glucose level in the
sample of tear fluid is associated with this signal [185]. Optical sensing,
on the other hand, utilizes the interaction between glucose and specific
K.V. Jarnda et al.
Fig. 1. An example of the electrochemical method and the subsequent signal processing used in the glucose concentration detecting system. Reprinted from Villena
Gonzales et al. [205].
Fig. 2. Illustration of how glucose oxidase converts glucose to gluconic acid.
Adapted from Kornecki et al. [111].
optical properties or molecules [224]. For instance, some optical sensors
that are not enzymatic can take advantage of the surface plasmon
resonance (SPR) phenomenon [224], where the presence of glucose al­
ters the refractive index of a sensing surface, resulting in a measurable
change in the light reflection or transmission [123].
The use of a fluorescent molecule to identify glucose in the tear fluid
is based on the fluorescence quenching principle, which is another op­
tical technique [51]. The fluorescence of the molecule is quenched in the
presence of glucose, and the amount of quenching is proportional to the
amount of glucose present [51]. Comparing these non-enzymatic tear
glucose sensors to enzymatic ones reveals benefits, including increased
stability, lower price, and extended shelf life [174]. However, they have
several drawbacks, including the possibility of drift over time, low
sensitivity, and sensitivity to interference from other compounds present
in tear fluid [17]. To improve tear glucose monitoring for diabetes pa­
tients, researchers are still investigating and developing new
non-enzymatic sensing approaches. Agostino et al. used reflex tear
samples to analyze their glucose level in real-time to show the CuO µPs
glucose sensor’s applicability to actual samples [174]. After being given
0 µM and 300 µM glucose supplements, two tear fluid samples were
examined in triplicate. During chronoamperometry tests, a 15 µL aliquot
of tear fluid was infused in 1.5 mL of support electrolyte (dilution 1:100)
[174]. Both the standard addition method and determining the actual
concentration from the calibration curve were used to quantify the
glucose [23], taking into account the dilution of 1:100 done when
spiking. In the first instance, extrapolating the calibration line to zero
response yielded a concentration of 4.125 mM for the spiked tear sam­
ple, as shown in Fig. 3a. Another study conducted by Han et al. [77] first
used the sensor system embedded with the glucose sensor and DES
sensor (W/O). To represent the normal and DES circumstances in
Fig. 3b, a tear film of two different thicknesses was created using an
artificial tear fluid. The glucose concentrations were adjusted for each
layer in order to calibrate the system’s glucose sensors. Both sensors in
the sensor system absorbed enough tear fluid when in contact with the
simulated tear film to perform measurements.
3
Sensors and Actuators: A. Physical 363 (2023) 114778
The glucose sensor demonstrated a highly linear relationship be­
tween the electrical current values and glucose concentration, as shown
below (Fig. 4A), irrespective of the tear film thicknesses (Fig. 4C). Based
on these findings, they came up with the equation below to calibrate the
glucose level in tears for in vivo testing: Tear glucose concentration for
in-vivo testing:
Glucose concentration (mg/dl) = 0.59(mg/dl Na) × Current (nA) -
3.76 (mg/dl).
Han et al. [79] evaluated the infiltration length for the DES sensor
(W/O) in the following four scenarios: DES film and low glucose con­
ditions, DES film and high glucose conditions, normal film, and low
glucose conditions (0, 0.9, and 1.8 mg/dl), normal film and high glucose
conditions (9 and 18 mg/dl), and normal film and DES film conditions.
Fig. 4B demonstrates that the infiltration length from the DES film was
statistically substantially shorter than that from the conventional film.
But for each video, there was no discernible difference between the two
distinct glucose circumstances, which correspond to tear fluids with
low/normal and high blood glucose levels [212]. This finding indicates
that there may be a threshold between the amounts of tear fluid
collected under the normal and DES situations.
2.2. Detection of non-enzymatic glucose in saliva
Analytes that get into the blood through the saliva can tell you about
your mood, hormones, metabolism, and nutrition [238]. Therefore,
saliva has been considered a fluid substitute for non-invasive glucose
monitoring. A healthy person’s blood glucose level ranges from 0.23 to
0.38 mM, while a diabetic person’s ranges from 0.55 to 1.77 mM [5].
Although salivary and blood glucose levels are related, little is known
about this connection [114], making it more challenging to connect
salivary concentration to therapeutic activity. The capability of
non-invasive saliva collection is one of the many benefits saliva offers for
diagnostic purposes [34].
Patients self-puncture at least 1800 times a year, according to Gro­
chowska et al. [70], which may encourage the development of
K.V. Jarnda et al. Sensors and Actuators: A. Physical 363 (2023) 114778

Fig. 3. (a). Detection of glucose in real tear sample using the standard addition method. (A) Chronoamperometry measurements with tear sample after supple­
menting it with 0 µM and 300 µM glucose. (B) Linear fit and extrapolation of the calibration line to zero response [174]. (b) A) Schematic description of the setup. (B)
Thickness profiles of tear films prepared under two different flow rates, simulating the normal and DES conditions, respectively [77].

inflammation at the injection site. Therefore, it is crucial to continue
research into the possibility of regulating the amount of glucose in the
human body while retaining the accuracy of blood tests by evaluating
fluids other than blood. In the research work of Grochowska et al.,
measurements were made in a deaerated NaOH solution at various
glucose concentrations (0.006–45 mM) in order to first assess the
sensitivity of the best samples among materials with varied Au film
thicknesses. In Fig. 5a, only a few samples out of the more than 30
registered cycles are included for clarity. The oxidation peak current
density registered in the reverse scan increases gradually with the
increasing glucose content in the electrolyte. Based on the oxidation
peak current, the current density versus concentration j vs. c relation
was graphically shown in Fig. 5b. At low glucose levels (0.05–6 mM), a
linear relationship between glucose concentration and current density is
discovered. This relationship has a sensitivity of 540 μA/cm2 mM. The
low detection limit of 10 μM was established using the relationship
3 SD/slope, where SD stands for standard deviation, and the slope is
derived from the linear regression model. This range corresponds not
only to the glucose concentration in blood (3.9–7.1 mM for healthy,
non-diabetics) but also to its levels in other human body fluids such as
saliva (0.008–0.21 mM), sweat (0.277–1.11 mM), and tears
(0.1–0.6 mM) [70], and interstitial fluid in which glucose concentration
is two orders of magnitude lower than in blood [180].
Moreover, the linear response can also be observed in the upper
concentration range (6–45 mM) with a sensitivity of 185 μA/cm2 mM.
This relationship overlaps with elevated physiological levels in the
blood (up to 11 mM) that can be related to prediabetes states and with
abnormal glucose levels (> 11 mM), which are registered for diabetics.
It should be underlined that the above-mentioned diagnostic criteria for
blood are set for a 2 h glucose toleration test [19]. When compared to
non-enzymatic sensors and other glucose sensing parameters for various
sensing electrodes in Table 1, the material proposed here shows
improved performance both in terms of sensitivity and limit of
detection.
Xu et al. [226] developed a graphene-based nano-sensor that was
applied to tooth enamel and printed on water-soluble silk to detect
germs (Fig. 6 A). The working electrode of the mouth guard sensing
platform is shown in (A) and (B) with an immobilized lactase enzyme.
Fig. 6B, taken from Zafar et al. [237], shows a non-invasive continuous
glucose monitoring tattoo on a dental platform. (A) Bio-transfer of the
sensor onto the tooth enamel; (B) Magnified image of the sensing unit
using a wireless readout method; (C) Magnified picture; and (D)

4
K.V. Jarnda et al. Sensors and Actuators: A. Physical 363 (2023) 114778

Fig. 4. In vitro evaluation of the sensor system. (A) Plots between the measured electrical current and known glucose concentrations. (B) Infiltration length of
collected tear fluid measured under in vitro simulated conditions. * ** * Statistically significantly different from all normal films (Student’s t-test; P < 0.0001) [78].
(C) Plots between the measured electrical current and known glucose concentrations in normal and DES films were obtained under in vitro environments [78].

Fig. 5. Electrochemical response of 50 nm Au-TiO2NTs to successive addition of glucose into 0.1 M NaOH (A) and corresponding relation between oxidation current
density vs. glucose concentration with calibration curve (B) [70].

Self-assembly of dangerous bacteria bound by peptides on the nano
transducer surface. This detecting tooth doesn’t require a power source
or any additional connections, thanks to the resonant coil inside. Anti­
microbial peptides are assembled on a single sheet of graphene in the
system, enabling bio-selective identification of bacteria at the cellular
level. Sun et al. [192] created a disposable electrochemical
saliva-glucose microfluidic system. The glucose-sensing chip in gluc­
ometers has working, counter, and reference electrodes [192]. Gold
nanoparticles enhanced the adhesion and signal sensitivity of the
complex device’s single-walled carbon nanotubes, which conveyed
glucose oxidase electrons, while chitosan sheets immobilized the
enzyme [192]. In healthy saliva, this equipment found glucose con­
centrations of 0.017–0.8 mM [192]. The field of non-enzymatic glucose
detection in saliva and tears holds great promise for improving diabetes
management. Future researchers should prioritize enhancing sensitivity,
reducing interference, and designing user-friendly, accurate sensors.
Collaboration with healthcare professionals and the integration of smart
technologies are key to the successful implementation of these methods

5
K.V. Jarnda et al. Sensors and Actuators: A. Physical 363 (2023) 114778

Table 1
Comparison of recent development of saliva/tears range in non-enzymatic glucose sensors and other glucose sensing parameters for various sensing electrodes.
Electrode Materials Glucose LOD (μM) Linear Applied Pros Cons References
Sensitivity Detection potential
(μA mM− 1 Range (mM) vs Ag/AgCl
cm− 2) (V)

Sandwich nanoporous 1950 1 0.1–6 0.58 Potential for high aspect Complex fabrication, [124]
framework decorated ratio, improved electrical limited scalability, limited
with vertical CuO conductivity, and to specific applications,
nanowire arrays enhanced surface area and and complex fabrication
mechanical stability
CuxO/Cu nanowire array 1210 10 0.01–7 0.6 Optoelectronic properties, Oxidation and degradation, [60]
large surface area, limited material
compatibility with compatibility, sensitivity to
microfabrication, and environmental conditions,
photocatalysis toxicity
Nanospindle-like Cu2O/ 2143 0.2 0.0005–2.5 0.4 Biocompatibility, Fabrication complexity, [143]
straight multiwalled improved mechanical chemical breakdown due to
carbon nanotube hybrid strength, high surface exposure to oxygen,
area, and photocatalytic restricted usability, and
activity compatibility of substances
Cu2O nanoparticles on 185 0.05 0.1–6 0.6 Compatibility with living Complexity in the [223]
reduced graphene oxide organisms, improved manufacturing process,
electrical conduction, responsiveness to changes
significant surface area, in environmental factors,
and enhanced mechanical restricted range of uses,
resilience and chemical reaction with
oxygen leading to
deterioration
Saliva Filter paper strip for - 1230 0.50–74.9 - Non-invasive collection, Variable sample saturation, [176]
salivary analysis easy and cost-effective, analyte variability,
suitable for remote potential contaminants,
sampling, and and limited applicability
quantitative analysis
Cu/Cu2O hollow 33.63 0.05 0.22–10.89 0.45 Unique structure, Challenges in the [195]
microspheres catalytic activity, manufacturing process,
photocatalysis, and material limitations, and
enhanced conductivity sensitivity to
environmental conditions
Graphene wrapped Cu2O - 3.3 0.3–3.3 0.6 Improved stability, Limited applications and [130]
nanotubes chemical responsiveness, material compatibility
increased electrical
conductivity, and
compatibility with
biological systems
Three-dimensional Cu 97.9 3.1 0.007–4.5 0.55 Customization, stability, Relatively expensive, [57]
foam-supported single- fast response, and sensitive to variations in
crystalline mesoporous specificity environmental conditions,
Cu2O nanothorn arrays and potential toxicity
Au NP/CeO2 513.72 1.83 0.02–0.6 0.9577 Constrained adaptability Intricate Production, [100]
and customizable constrained flexibility, cost
properties considerations
Rod-like PtAu-MnO2 58.54 20 0.1–30.0 0 High conductivity, precious metal cost, [191]
binary nanocomposites tailored properties, and possible harmful effects,
on graphene paper synergistic effects and vulnerability to
changes in the
surroundings
Single-walled carbon 26.6 5 0.1017–1.11 0.2 Rapid response, Cost, interference, and [132]
nanotubes and miniaturization, limited versatility and
multilayers of chitosan, customizability, and stability
gold nanoparticles, and elevated sensitivity
glucose oxidase on chip
saliva nano-biosensor
Porous NiO 2432 0.084 0.005–0.825 0.995 Chemical reactivity, Sample preparation, [26]
nanostructures energy storage, sensing potential toxicity, and
capabilities, and high limited specificity
surface area
Tears
Nafion/CuO-μ Ps/Au 850 2.99 0.003–0.7 0.40 Enhanced electrochemical Complex fabrication, [174]
properties, stability in limited versatility,
aqueous environments, potential toxicity, and
and sensitivity for tear sample collection and
fluid analysis handling
GR-CNT-ZnO-GOx 5.36 4.5 0.01–6.5 0.73 Rapid response, possible Irritations, expensive, and [103]
( ± 0.072) compatibility with sensitive to environmental
biological systems and factors such as
(continued on next page)

6
K.V. Jarnda et al. Sensors and Actuators: A. Physical 363 (2023) 114778

Table 1 (continued )
Electrode Materials Glucose LOD (μM) Linear Applied Pros Cons References
Sensitivity Detection potential
(μA mM− 1 Range (mM) vs Ag/AgCl
cm− 2) (V)

durability in watery temperature, humidity, or

surroundings pH
Graphene/CNT 53.89 3.99 × 10–7 m 0.004–5 1.9 Potential for continuous Potential interference, [239]
composite monitoring, costly, and vulnerability to
miniaturization, and changes in the
immediate response surroundings
GR-MWNTs/AuNPs 0.695 4.1 10 0.53 High electrical Material suitability, [235]
conductivity, enhanced susceptibility to
catalysis, a large surface environmental changes;
area, stability, constrained adaptability;
customizability, and and cost-related factors
potential suitability
5 2
Cu–Gox/CNTPE (2.544 2.0 × 10− 1.20 × 10− 0.32 Potential use in diabetes Collection and [87]
± 0.008) management, heightened management of samples;
sensitivity, compact Susceptibility to
design, and flexibility for environmental factors;
customization financial implications, and
the potential for external
influences.

Fig. 6. (A) Shows saliva non-invasive continuous lactate detection and (B) Illustrates glucose-sensing tattoo printed onto a tooth platform.
Modified from Xu et al. [225].

in clinical practice and daily life.
3. Electrochemical principles of glucose sensors
The biosensor industry must prioritize the creation of the most
effective glucose sensor. Biosensors for glucose have been manufactured
utilizing electrochemical, colorimetric, conductometric, optical, and
fluorescence spectroscopy [25]. Electrochemical glucose sensors have
dominated the market for the past 40 years due to their high sensitivity
and selectivity. Less costly, quicker, more stable, and with a lower
detection limit are electrochemical approaches. A single molecule can be
detected by a number of fluorescence-based in vivo glucose monitoring
systems; however, none of them are helpful for the management of
diabetes [211]. Electrochemical glucose sensors can be put into three
groups based on how they measure glucose: amperometric, impede­
metric, conductometric, and potentiometric.
3.1. Amperometry glucose sensor
Amperometry is sensitive due to constant voltmeter and linearly
dependent target concentration, while GOx oxidizes glucose on noble
metal electrodes [16]. An amperage-measuring biosensor consists of two
or three electrodes, with the working electrode and the reference elec­
trode being the primary components. The two-electrode approach has
limited utility in biosensors because large potential drops at high current
flows make potential control difficult [127]. In glucose sensors, the third
electrode, a high-surface-area auxiliary counter electrode, increases the
current between the counter and working electrodes as voltage is
applied. While voltage is still delivered between the working and
reference electrodes, the third electrode is typically used as an auxiliary
counter electrode with a large surface area to maximize the flow of
current between the counter and working electrodes. Depending on the
mechanisms of electron transfer, there are three distinct forms of glucose
oxidation known as first, second, and third generation glucose sensors
[119]. At a first-generation amperometric enzyme electrode, the
following three amperometric signalling modes can be used for glucose
assay: cathodic detection of O2 and H2O2 [74] and anodic detection of
H2O2. Compared to the other two methods, anodic detection of H2O2 to
replenish O2 is unquestionably the most effective method for enhancing
the enzymatic reaction cycle [73].
3.2. Impedimetric or conductometric glucose sensor
Impedimetric measurements are less commonly implemented than
amperometric and potentiometric measurements. Electrochemical
impedance spectroscopy (EIS) can reveal the electrical double-layer
structure of a modified electrode; however, it takes time to record an
impedance spectrum over a broad frequency range [141]. The dielectric
spectra at MHz frequencies are unaffected by glucose [109]. Due to the
fact that conductance is the inverse of resistance, impedimetric bio­
sensors are frequently referred to as conductometric biosensors [179].
Jeon et al. developed a glucose quantified technique using EIS results,
immobilizing GOx with MPA and using hydroquinone as an electron
mediator. EIS measurements indicate that as glucose content rises, H2Q

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K.V. Jarnda et al.
Fig. 7. (A) Diagram illustrating the structure of an electrochemical glucose sensor.(B) The evolution of invasive glucose sensors to non-invasive electrochemical
glucose monitoring systems.
(a) Reproduce from Bandodkar et al. (2018). (b) Adapted from Lee et al. [118].
oxidation increases diffusion current density, thereby decreasing charge
transfer resistance (Rct). The simple, non-destructive method measured
glucose dynamically with a linear sensor response (1/Rct) based on
glucose in solution [98].
3.3. Potentiometric glucose sensor
Potentiometry measures glucose concentrations above 10− 5 M at
physiological levels, determining the reference-indication electrode
potential difference at zero current flow. The ideal nonpolarizable
reference electrode has a stable potential, whereas the potential of the
indicator electrode fluctuates in response to the analyte concentration.
Target analyte concentrations have a logarithmic effect on the zero
current potentials between these electrodes [173]. Commercial appli­
cations for enzyme-based glucose biosensors exist, but non-enzymatic
glucose sensors can overcome GOx’s volatility and short shelf life,
which limits their repeatability. These glucose sensors detect glucose on
the surface without using enzymes such as glucose oxidase [124,197].
Enzymatic sensors used in healthcare, initially performed poorly on
low-selectivity devices due to their potential resemblance to enzymes in
the substrate, electrodes, and materials used [107,215].
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Sensors and Actuators: A. Physical 363 (2023) 114778
4. Non-enzymatic glucose detection method
Due to a larger amount of blood sample and the removal of red blood
cells, commercial glucose sensors quickly switched from photometric to
enzymatic electrochemical detectors [115]. Enzyme-based glucose
sensors currently dominate the industry, but non-enzymatic sensors are
being developed to address issues with traditional enzymatic sensors.
Most commercially available disposable glucose sensors are created
using well-calibrated combinations. Several ways to keep an enzyme
from moving, such as crosslinking, direct adsorption, and sol-gel
adsorption, have been written about and used in the cocktail method
[18]. Electro-polymerization can also collect enzymes in mixtures with
monomers, cross-linking agents, and enzymes [207]. This method is
intriguing due to the one-pot immobilization of the enzyme and elec­
trochemical control of the enzyme layer thickness. However, because
the sensitivity of glucose sensors is largely reliant on the activity of the
immobilized enzyme, reproducibility continues to be a key issue in
quality control. Glucose sensors that directly oxidize glucose on the
electrode surface to produce electrical current should be able to alleviate
the oxygen restriction problems. As oxygen’s thermodynamic reduction
potential is higher than glucose’s oxidation potential and the reduction
reaction of the oxygen molecule is electro-kinetically slow on most
electrode surfaces, applying the right potential should minimize this
K.V. Jarnda et al. Sensors and Actuators: A. Physical 363 (2023) 114778

Fig. 8. (A) An overview of the first, second, and third generations of glucose oxidation sensors. (B) Demonstrates the operation of an amperometric glucose sensor
that is oxygen dependent [96]. (C) The working electrode in the MOSFET-based potentiometric glucose detection is an extended-gate functionalized ZnO nanowire,
and the reference electrode is an Ag/AgCl. (D) (a) cross-sectional and (b) top views of the potentiometric arrayed glucose biosensor, respectively. (c) Optical image of
the glucose biosensor based on Nafion GOx/GO/AZO and a potentiometric array [36].
(a) Adapted from Dhara and Mahapatra [41]. (b) Adapted from Ridhuan et al. [171].

Table 2
Studies on Non- enzymatic Sensing approaches to improve the detection of glucose.
Sensing Mode of Detection Linear Detection Range (mM) Detection Limit (μM) Specimen References

CuO/PCL@PPy/ITO Electrochemistry 2 μM - 6 0.8 Saliva [33]

IrO2 @ NiO nanowires Electrochemistry 0.5 μM - 2.5 0.31 Saliva [46]
rGO modified Nb2O5 Electrochemistry 1–10 1 Tears, Urine, Saliva [139]
Pt/Ni@NGT paper based device Calorimetry 0.1–50 1 Tears, Saliva [216]
Au NP/CeO2 Spectrometry 0.02–0.6 1.83 Saliva [216]
Bi-enzyme based Electrochemistry 0.05–1.5 3 Saliva [88]
Os-complex thin film
SWCNT/Au nanoparticle/GOx Electrochemistry 0.017–0.81 5.6 Saliva [172]
Au-CuO NRs Electrochemistry 0.005–1.325 0.17 Saliva [27]
CuO PN Electrochemistry 0.005–0.225 0.41 Saliva [28]
PBS-Cu-Ni Electrochemical 5 nM - 20 mM 4.2 nM Saliva [68]

9
K.V. Jarnda et al.
interference. Because of the inherent characteristics of enzymes, the
most common and significant concern is the stability issue with enzyme
sensors, as shown in Table 2. Enzyme-based glucose sensors are sus­
ceptible to temperature or chemical deformations throughout the course
of their development, storage, and use [158].
4.1. Non-enzymatic glucose sensing mechanisms
4.1.1. Mutarotation of glucose
There are five different isomers of the aldohexose that makes up D-
glucose in nature (see Fig. 9(a)). The open-chain aldehyde molecule, g-
glucose, is hydrolyzed by an acid to generate a five-membered ring
known as furanose or a six-membered ring known as pyranose. Based on
the cis-trans configuration of the hydroxyl group attached to C-1, the
two cyclic glucoses can be divided into anomers. Even though the
numbers can be a little different depending on the research, it is known
that the ratio of α glucopyranose to β glucofuranose and to γ glucose in
an aqueous solution, at equilibrium is about (α and β): γ-glucose = 38:
62:< 1.0:< 0.1.
Through the mutarotation mechanism, pyranoses, which make up
most of the glucose solution, can be broken down through an open-chain
g-glucose intermediate and brought back into balance. At room tem­
perature, the mutarotation process can take up to 2 h to reach equilib­
rium [168]. The equilibrium and isomeric ratio of the mutarotation are
also significantly influenced by the physico-chemical environment,
including pH [128], temperature [162], and coexisting ions [52]. For
instance, after equilibrating for 5 h at 2 ◦ C, D-glucose solution has the
following composition: α: β = 56: 44 at pH 1.0,: β = 27: 73 at pH 7.0,
and α: β = 22: 78 at pH 13 [64]. The rate-determining step in the
non-enzymatic oxidation of glucose is the removal of the hydrogen atom
at C-1. It is much easier to remove the hydrogen atom from α -gluco­
pyranse than from the other sugar. This is like the way the enzymes GOx
and GDH, which oxidize glucose, move. The C-1 hydrogen atom is
rapidly activated as a result of the hydroxide group’s high acidity (pKa =
12.3, higher than that of aliphatic alcohols, which is pKa = 16). Also, the
Fig. 9. (A) The molecular forms of several D-glucose isomers. The composition ratio of an equilibrium aqueous glucose solution at standard temperature and pressure
is shown in parenthesis. For emphasis on stereochemistry, the functional groups at carbon atom 1 are highlighted. (B) The overall process of oxidizing glucose.
Redrawn from Reghunath et al. [170].
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Sensors and Actuators: A. Physical 363 (2023) 114778
axial hydrogen of the α -glucopyranse is much easier to reach than the
equatorial hydrogen of the form [117]. Regardless of the anomeric
forms, dehydrogenated glucose is quickly oxidized into
glucono-d-lactone, which is then rapidly hydrolyzed at a rate constant of
10 − 3 s− 1 to produce gluconic acid [67]. The overall non-enzymatic
process of glucose oxidation is depicted in Fig. 9(b).
4.1.2. Incipient hydrous oxide adatom mediator Model
Burke [21] also mentioned another concept, which is known as the
“incipient hydrous oxide adatom mediator (IHOAM) model”. The elec­
troactive substance’s low structural stability on the electrode surface led
to its reactivity [95]. The immature hydrous oxide (M-OH ads), which is
regarded as a crucial mediator for the electro-oxidation processes of the
glucose molecule in the electrolyte, is created when the hydroxide ion
(OH) from the electrolyte adsorbs on the electrode. The glucose mole­
cule is then oxidized by the M-OH ads mediator, which also returns the
M-OH ads to their metallic state. The oxidation process (the red region on
the left in Fig. 10 (A)) and the reduction reaction may therefore be
distinguished from the overall reactions (the blue area on the right). The
oxidation of glucose molecules may be seen in the red area (left side).
Electroactive materials go through multiple cycles of oxidation and
reduction processes in the middle. Lastly, the appropriate reduction
reactions are shown in the blue area (right side). There are various kinds
of non-enzymatic electrocatalysts; they include noble metals like plat­
inum and gold, as well as metal oxides, alloys, and compounds con­
taining carbon. Adsorption of the analyte to the electrode surface, which
commonly involves d-electrons and d-orbitals of the metal substrate and
results in the formation of a bond with the adsorbate, is a typical step in
the electrocatalytic process [177]. The adsorption process depends
heavily on the form of the electrode. This suggests that the organic
analyte’s chemisorption and the hemiacetalic hydrogen’s removal occur
at the same time [157]. The idea of active transition metal sites provides
a satisfactory explanation for the adsorption process that takes place on
electrode surfaces. The kind of electrode material used in non-enzymatic
sensors significantly affects how glucose is directly electro-oxidized.
K.V. Jarnda et al. Sensors and Actuators: A. Physical 363 (2023) 114778

Fig. 10. (A) The general reactions of the process are demonstrated by an example of how glucose is detected using the Incipient Hydrous Oxide Adatom Mediator
(IHOAM) on non-enzymatic electrodesModel (B) for the early mediator between hydrous oxide and adatom. The rapid electro-oxidation of glucose into glucono-d-
lactone is thought to be facilitated by the electrode’s reactive hydrous oxide (OH ads) layer. (C) The redox reactions of many organic compounds as a result of reactive
hydroxide species on the electrode surface (OH ads) formed during electrocatalysis.
(reproduced from Chiu et al. [35].

Electrocatalytic processes are therefore essential for the oxidation of
glucose.
4.1.3. Direct glucose oxidation via metallic redox centres
Electrocatalytic processes are mostly done by a process called
adsorption, in which reactant molecules stick to the active parts of the
electrode. The adsorption mechanism can be changed by many things,
such as favourable electronic states at redox centers, empty d-orbitals at
transition metal centres, or many flaws in non-metal-based catalysts.
After the reactant has been adsorbed, bonds break down and in­
termediates are produced. The reaction product desorbs from the elec­
trode surface as the contact between the product and the electrode
reduces and the oxidation state of the redox centre changes. This reac­
tant adsorption and desorption process on the electrode is referred to as
the chemisorption model [241]. During chemisorption-based glucose
oxidation, the amount of chemical interaction between C-1 and its
hydrogen atom and the electrode surface rises, which causes C-1 to
dehydrogenate and adsorb to the electrode surface (Fig. 11a). Following
electro-oxidation of the adsorbates, glucono-lactone is produced, which
is then transformed into gluconic acid through a number of reaction
pathways depending on the pH [99,153]. The electrocatalytic produc­
tion of surface-bound reactive hydroxide species (OH ads), in which
surface hydroxide radicals directly oxidise the reactants, has a big effect
on the redox reactions of small organic molecules.

Fig. 11. (A) Chemisorption model glucose oxidation mechanism from Naikoo et al. [153], (B) The activated chemisorption model is linked to the electro-oxidation of
glucose at metal-active nuclei.
Redrawn from Chou et al. [36].

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K.V. Jarnda et al. Sensors and Actuators: A. Physical 363 (2023) 114778

Fig. 12. The number of articles published in the last 5 years on non-enzymatic electrochemical and electrochemical glucose sensors (date acquired in September
2023: PubMed).

Fig. 13. (A) Optical micrographs of a bare Au-modified microneedle electrode array (MNEA) showing a length of 599.61 µm and a width of 100.16 µM (Naikoo et al.,
2021). (B) Schematic illustration of the mesoporous needle-like Co3O4 nanowires for immobilizing the redox enzyme GOx by adsorption and entrapment. (C)
Schematic mechanism for non-enzymatic glucose sensor fabricated based on CuO spheres composed of finely arranged nanobricks modified silver electrode. (D)
Schematic representation showing Fe3O4-rGO-gelatin nanocomposite preparation on GCE sensing electrode.
(B) adapted from Naikoo et al. [153]. (c) reprinted from Chen et al. [31]. (d) adapted from Iftikhar et al. [92].

4.1.4. Chemisorption model and oxidation of glucose in this electrolyte.

According to the chemisorption model [99], electrocatalysis often
occurs when the analyte is adsorbed on the electrode’s active sites,
where a strong bond is created with the adsorbate. Reactants that have
been adsorbed form intermediates by rupturing bonds. Using the
chemisorption model, products desorb from electrode surfaces as
electrode-product contacts deteriorate [200]. The glucose molecule in­
teracts with C-1 and its hydrogen atom, leading to electrode adsorption
5. Recent prospects in non-enzymatic glucose sensor
Non-enzymatic glucose sensing is a cheap and rapid technique that
relies on the direct electrochemistry of glucose [56]. Glucose oxidation
or reduction is the foundation of non-enzymatic glucose sensing [215].
There are three significant drawbacks to direct glucose oxidation on

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K.V. Jarnda et al. Sensors and Actuators: A. Physical 363 (2023) 114778

Fig. 14. (A) Cyclic voltammetry graph displaying three potential regions where glucose is electro-chemically oxidized at a Pt electrode [20]. (B) Schematic rep­
resentation of nickel/copper/carbon nanotubes nanocomposite electrode. (C) The mechanism involved in the activation of Ni-Pd@AC/GCE using NaOH solution. (D)
SEM images of Pt coatings obtained at different deposition times (bars: 200 nm) [54]. (E) The oxidation reduction of glucose at the peak of 0.8 V is illustrated in the
cyclic voltammetry curve of the ZnO nanorods electrochemical sensor where (a) is in the absence of glucose; (b) is at 1 mM glucose concentration; and (c) is at 2 mM
glucose concentration.
(B) Adapted with permission from Ammara et al. [11]. (C) Printed from Koskun et al. [112]. (c) Adapted from El Golli et al. [53].

noble metal electrodes: (1) reduced glucose sensitivity because con­
ventional electrodes have delayed glucose electro-oxidative kinetics; (2)
low selectivity; and (3) decreased electrode activity because of chloride
ion (CI) contamination. More glucose can directly reach the electrode by
expanding its surface area, which enhances sensitivity and selectivity.
To do this, nanomaterials are being researched [91,149]. Since they are
widely used to develop novel non-enzymatic glucose sensors, noble
metal alloys such as Au, Co, Cu, Fe, Mn, Ni, Pd, Pt, Rh, and Zn are
particularly significant [140,149]. Because the hydroxyl groups (OH) on
the electrode surface prevent chloride from sticking to the surface, under
alkaline circumstances, there can be a lift on the ion contamination re­
striction [62]. The non-enzymatic glucose oxidation catalytic process
includes the hemiacetalic hydrogen atom abstraction, which occurs
simultaneously with the adsorption of the organic species [183]. This
method is assumed to be the rate-determining step in the catalytic
process of glucose electro-oxidation.
5.1. Metal Alloy-Based Glucose Sensors, such as: COFs, MOFs, and other
materials
5.1.1. Covalent based framework for biomolecules sensing of glucose
A number of significant disorders can be identified by the presence of
certain tiny biomolecules, including glucose, uric acid, glutathione
(GSH), and others. New materials for simple detection of tiny bio­
molecules are being developed, enhancing medical diagnosis and
treatment, in line with the growing demand for portable and wearable
medical devices. Recently, certain COFs or COF-based hybrid materials
were created and produced for the detection of tiny biomolecules such as
GSH, glucose, and uric acid (Table 3). Blood glucose level is a crucial
indicator for diabetes diagnosis, and various methods can be used to
measure glucose levels [102]. J. N. Wang and colleagues created a COF
(Fe COF) in 2018 that mimics peroxidase and is based on Fe-porphyrin
[122]. Similar to the COF employed by Gua, Y, and colleagues [209], Fe
COF can be used to quantitatively measure (H2O2) by producing ox TMP
that is blue in colour. A glucose sensor was developed by combining
GOx, non-enzymatic glucose oxidation, and Fe COF-catalyzed ox TMP
synthesis, with a detection range of 5–350 µM and a 1 µM threshold.

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Table 3
COFs or COF-based hybrid materials for detecting glucose and other small biomolecules in recent years.
Analyte Hybrid Names Binding Site Detectable Signal Linear Detection Range (mM) Detection Limit (μM) References

GSH COF− 300 − AR - Chromism 1–15 µM 1.0 µM [102]

GSH Py− TT COF - Chromism 0.4 − 60 µM 0.225 µM [102]
Glucose Fe- COF GOx Chromism 5–350 µM 1.1 µM [122]
Glucose COFDHTA-TTA GOx Electrochemical signal 0.60 µM–6.0 mM 0.38 µM [228]
Glucose Fe− PorCOF GOx Chemiluminescence 0.01–10 µM 5.3 nM [228]
Glucose COFHD–GOX GOx Chromism 5–2000 µM 0.54 µM [246]
Uric Acid COF− DC− 8 Hydroxyl, triazine Electrochemical signals 5.0–25 µM, 25–250 µM 0.77 µM [236]
Ascorbic Acid COF− DC− 8 Hydroxyl, triazine Electrochemical signals 30–180 µM, 12.0 µM [236]
0.18–1.5 µM
Dopamine COF− DC− 8 Hydroxyl, triazine Electrochemical signals 1.0–6.0 µM, 0.25 µM [236]
8.0–50 µM

Many COF-based glucose sensors with enhanced features have been
created on the basis of this theory [228].
5.2. Metal organic frameworks based / metal oxide-based on non-
enzymatic glucose sensors
In addition to adding carbon nanomaterials to increase conductivity,
another way to improve metal organic framework (MOF) performance is
to load electrochemically active metal nanoparticles into their channels
to make composites of MOF, metal, and metal oxide [84]. Metal nano­
particles are evenly distributed in MOFs, as demonstrated by Duan et al.
[49] in the dehydrogenation of hydrazine monohydrate in alkaline so­
lution using NiIr nanoparticles. Two types of metal-ZSM− 5 electrodes
were utilized for the electrocatalytic oxidation of glucose and methanol,
namely transition-metal ion exchange form (M-ZSM− 5) and
nano-crystalline form (M-Nano-ZSM− 5) [69]. MOFs with porous metal
oxides and MO@C have been used as sacrifice templates to create
high-performance sensor electrodes, providing a carbon coating, and
preventing agglomeration due to their homogeneous dispersion of
metallic species over carbon materials. MOF-generated materials are
ideal for self-supported electrodes due to their unique structure and
potential for various electrochemical applications due to their
outstanding structural characteristics. Table 4 presents typical examples
of MOF-produced materials utilized in non-enzymatic glucose sensing
applications.
5.3. Other metal alloy-based glucose sensors materials
Many metals, in particular noble metals, have been studied as base
materials for the electrodes of non-enzymatic glucose biosensors [120].
Because of this, scientists now have a better understanding of how
glucose direct oxidation works, which shows that the mechanism is
completely dependent on the metal catalyst in the electrode [56,154].
Furthermore, as a result of advancements in material science, a number
of metal alloys and hybrid materials have been developed, allowing for
enhancements in properties over noble metals and metal oxides alone
[90,202]. This section discusses innovative non-enzymatic glucose
sensors made from metal alloys like Au, Co, Cu, Fe, Mn, Ni, Pd, Pt, Rh,
and Zn, widely used in sensor development.
5.3.1. Gold (Au) metal oxide based glucose sensors
Gold is a well-researched electrode material that, in contrast to other
noble metals, generates a high glucose oxidation current in neutral or
alkaline conditions [175]. Au has been extensively researched in glucose
fuel cells, serving as the standard electrode material for glucose oxida­
tion. However, the oxidation mechanism remains unknown, necessi­
tating further study. The anodic scan of the Au cyclic voltammogram
reveals only the double layer and Au oxide regions [245]. Previous
studies have suggested a connection between the oxidation process and
the existence of surface oxides such as Au (OH) [234]. Also, in the
double-layer region of the Au oxide region, glucose oxidation happens
more often than in the Pt oxide region, where the surface OH-ads layers
are made [91]. Low pH levels, according to the findings, only allow the
detection of oxidative current at potentials above the oxide region, but
high pH levels cause a spike in faradic current [150]. Many methods
have been used to create nano porous Au samples, such as electro­
chemical etching and dissolving [202,229], electrochemical deposition
[198], and thermal annealing [91,184], in order to lessen ion contam­
ination and interference with the sensor surface.
5.3.2. Cobalt (Co) metal oxide based glucose sensors
Scientists are testing co-based catalysts for glucose sensors that don’t
use enzymes. They are focusing on making compounds, analysing to­
pologies, and finding the best supports for active catalytic loading. Li’s
research group developed non-enzymatic glucose-sensing CoP nanorods
by oxidizing Co(OH)2, Co3O4, allowing glucose monitoring in alkaline
solutions without the need for enzymes [125], CoOOH, and CoO2 [232].
The valence of CoP in its orthorhombic crystal changed, and
sugar-activated alkaline CoP density functional theory (DFT) simula­
tions were used to explain how CoP works as an electrocatalyst for
glucose. The study validated the experimental finding that non-noble
transition metal compounds exclusively electrocatalyzed glucose at
high pH through the interaction of glucose, water, and OH- [129]. Guo
and colleagues developed high-performance glucose monitoring using
porous carbon-coated nano-like Co-S, which consist of small, ionically
bound molecules rather than transition metal oxides [72]. Using cobalt
phthalocyanine and CNTs, Pathak’s team developed an enzyme-free
amperometric glucose sensor [160]. Cobalt phthalocyanine is a poten­
tial electron mediator for enzyme-based glucose biosensors, but its
structural and electrical failures require conductive supports like CNTs,

Table 4
Non-enzymatic sensing approaches for glucose based on MOF-derived materials.
Catalyst Potential Interferences Real Sample Detection Method Sensitivity Linear Detection Range (mM) LOD (μM) References

Cu -MOF Sucrose, UA, AA, DA Urine DPV 89 0.06–5 10.5 [147]

Cu-MOF/ CPE AA, UA, GSH, NaCl - Amperometry - 0.005–2.8 1 [39]
[Cu3 (BTC)2] AA, UA, DA, KCl - Amperometry 549 0.125–2250 - [93]
Ni-MIL-77 AA, UA, DA, KCl Serum Amperometry 1.542 1–500 0.25 [169]
Ni-MOF NaCl, DA, UA, Urea Sweat Chronoamperometry 3.03 10–2000 1.16 [10]
Ni-BDC UA, AA Blood Amperometry 635.9 0.01–0.8 6.681 [112]
Ni-MOF/CNTs AA, Lactose Serum Amperometry 13.85 1–1.6 0.82 [244]
AgNPs/MOF-4(Ni) DA, Sucrose Blood Amperometry 1.29 0.3–2.312 4.7 [161]

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Table 5
Summarized electrochemical detection of Au, Co, Cu, Fe, and Mn metal alloy based on non-enzymatic glucose sensor.
Electrode Materials Linear Detection Range (mM) Sensitivity (μA mM− 1) Detection Limit (μM) References

Au alloy nanoparticles 0.08–9.26 22 4 [196]

Au Nano flowers 0.3–30 20 150 [247]
Au/TiO2 NTs 5–40000 170 2 [104]
AuNPs-MWCNT 0.02–10 19.3 2.3 [131]
AuNPs 0.001–5 47.2 1.0 [86]
Co4N nanosheets 0.6–10.0 1137.2 0.1 [133]
Co3O4 nanostructures 0.5–4.5 27.3 0.8 [126]
CoP nanorods/GCE 0–5.5 116.8 9 [108]
Co3O4 NFs 20–2040 36.25 0.97 [63]
CoNiCu Alloy 1.551–4.050 322 0.5 [65]
CuO/SWCNTs/ITO 0.05–1800 1610 50 [151]
CuO nanoleaves–MWCNTs 0–0.9 664.3 5.7 [82]
Cu-CuO NWs/GCE 0.1–12 122.73 0.05 [9]
CuCo carbon nanofibers 0.02–1.1 507 1 [203]
Cu2O 0–6 629 2.4 [101]
Fe2O3 0.05–18 105.75 12 [55]
Fe3O4 0.0001–0.125 673.3 0.1 [148]
Fe@ZnO < 18 - 12 [94]
Mn3O4 0.001–0.3295 0.026 0.5 [58]
MnO2 -28 33.19 - [230]

graphene, and high-conductivity metals [81].
5.3.3. Copper (Cu) metal oxide based glucose sensors
Copper-based nanoparticles are highly desirable for NEGS
manufacturing due to their high surface-to-volume ratio, sensitivity, and
selective performance. These parts are readily available, have improved
electrochemical properties, and make it simple to modify the copper
oxide structure of sensors [193]. Khoshroo et al. [106] reported a
number of significant parallels. Another study developed a NEGS based
on copper oxide nanoparticles that had good sensitivity, reproducibility,
a broad linear range up to 10 mM, and a broad linear range of 663.2 μA
mM− 1 cm− 2 [9]. A remarkable CuO hollow-sphere structure-based
sensor was created by Haghparas et al. [75]. This sensor had a good
linear range between 1 μM and 16 mM with a high sensitivity of 35.2
± 0.4 μA mM− 1 cm− 2 and a very low detection limit of 1 M [76].
5.3.4. Iron (Fe) metal oxide based glucose sensors
The most common type of transition metal is an electron mediator
(II), and Fe oxides (FeO, Fe2O3, and Fe3O4) are used in analytical studies
because metallic Fe is unstable in air. Fe3O4 nanostructures are often
used as magnetic materials for separation, medication delivery, and
magnetic resonance imaging. Mimetic peroxidase Fe3O4 NPs catalyze
hydrogen peroxide, and Fe redox directly electrocatalyzed glucose
oxidation [66,218]. According to Abrori et al. [4], Fe3O4 NPs supported
on CNTs were used for the non-enzymatic electrochemical detection of
glucose. In particular, the measurements were done by the authors in a
pH 7.0 phosphate buffer solution (PBS). A pair of redox peaks in the
potential range of − 0.5 to-0.2 V vs. Ag/AgCl were seen in the absence of
the analyte and were attributed to the Fe(III)/Fe(II) couple. In a different
work done by Abrori et al. [4], Fe3O4-based NEGS with high sensitivity
(4.67 μA Mm− 1 cm− 2) and a LOD of 15.70 μM were produced to elec­
trochemically detect glucose molecules. The sensor demonstrated high
sensitivity and selectivity for glucose molecules.
5.3.5. Manganese (Mn) metal oxide based glucose sensors
Due to its high specific capacitance and low cost, MnO2, a substantial
Mn oxide, is being studied a lot in the field of energy storage. Manganese
oxide (MnO2) is commonly used in electrochemistry and has been
explored in studies for the detection of glucose in supercapacitors and Li-
ion batteries [29]. Yin et al. [233] experiment revealed a sensor with an
improved sensitivity of 18.9 M− 1 cm− 2, enhanced by multiwalled car­
bon nanotubes. MnO2 and glucose molecules combine, forming 2MnO2
+ C6H12O6 and 2MnOOH + C6H10O6, respectively. Mn3O4 is the most
effective catalyst in NEGS [155].
5.3.6. Nickel (Ni) metal oxide based glucose sensors
Ni3+/Ni2+ and its derivatives have exceptional catalytic activity
under alkaline conditions, making them the most widely used non-
enzymatic electrochemical glucose sensors. To develop a surface rich
in Ni2+ for glucose sensing, Ni must be rearranged to form Ni(OH)2 at a
particular voltage [145]. Following the oxidation of glucose to + 3, Ni
(OH)2 lowers + 2 gluconolactone. Similar to cobalt-based sensors,
nickel-based sensors have high glucose electro-oxidation current den­
sities [134,186,187,38]. Nickel oxide (NiO) is a great material for a
glucose-detecting device because it is very stable and has a lot of cata­
lytic activity. Moreover, these sensors display enhanced characteristics
such as a quick response time, increased sensitivity, a reduced degree of
recognition, superior cyclability and stability, and a big linear recipro­
cation window for glucose concentrations [190]. CNTs, graphene [146],
and other carbon compounds, as well as new substrates [156], have been
loaded onto Ni-based catalysts for a long time. Ni (OH)2 nanosheets that
sense glucose were affixed by Wu et al. to free-standing 3D graphene
foam [219].
5.3.7. Palladium (Pd) metal oxide based glucose sensors
Palladium-based sensors have just recently been developed [214].
They made use of Pd, Mn, and rGO mixed metal alloy nanoparticles,
which could detect glucose molecules in an alkaline environment. The
hybrid sensor demonstrated enhanced electrochemical functioning
during sensing as well as higher sensitivity and selectivity toward
glucose molecules [214]. A related work discovered good palladium
properties, repeatability, and selectivity for glucose molecules [214].
Unique Pd nanosponge designs with a wide linear range of 1–18 mM and
a great sensitivity of 32 A mM− 1 cm− 2 were produced by Promsuwan
et al. [164]. The glucose sensor had a LOD of 2 M and was consistent
over time.
5.3.8. Platinum (Pt) metal oxide based glucose sensors
Because of its high catalytic activity and stability, Pt is a good elec­
trocatalytic electrode material, according to recent research [136].
Pt-based glucose biosensors are stable, sensitive, and repeatable, offer­
ing low LOD, quick reaction time, wide linear range, and high sensi­
tivity. These NEGS, with Pt-based electrodes, offer excellent stability,
enhancing sensor function and stability, according to recent studies [15,
32]. These sensors are 10,000 times more sensitive than regular sensors
at a normal pH [14].
5.3.9. Rhodium (Rh) metal oxide based glucose sensors
Several investigations on the detection of hyperglycemia utilizing

15
K.V. Jarnda et al. Sensors and Actuators: A. Physical 363 (2023) 114778

rhodium oxide-based sensors have been undertaken [201]. Rhodium Table 6

oxide nanocorals (Rh2O3 NCs) on a modified glass carbon electrode can Using a non-enzymatic glucose sensor, summarized electrochemical detection of
be employed for electrochemical glucose oxidation, as Dong et al. [45] Ni, Pd, Pt, Rh, and Zn metal alloys was performed.
have effectively shown. With a LOD of 3.1 M and 11.46 μA mM− 1 cm− 2, Electrode Linear Detection Sensitivity Detection References
this sensor displayed remarkable sensitivity. The scientists developed Materials Range (mM) Limit (μM)
this sensor based on a paper that outlined a two-step fabrication pro­ Ni/CNTs 5–7000 1433 2.0 [121]
cedure for Co3O4 nanofibers that were cast on the surface of a glass NiO NPs 1–110 5590 0.16 [144]
carbon electrode and connected to Nafion [45]. It was also shown that Pd NPs/ 10–5000 - 1.0 [83]
graphene
the Co3O4-NFs-Nafion/GCE-based NEGS worked well when
Pd NTs 5–10000 6.58 1.0 [208]
electro-oxidizing glucose in an alkaline environment. On the periodic Pt nanoclusters- 0.003–12 12.8 1.0 [210]
table, rhodium and cobalt are close to one another; therefore, Unmüssig MWCNTs
et al. [201] chose to predict their correlation. The linear range, sensi­ Pt NPs- 0.003–8.2 16.1 1.0 [8]
tivity, and detection limit for various electrode materials such as Ni, Pd, MWCNTs-
PANI
Pt, Rh, and Zn are shown in Table 6. Rh2O3 NCs - 11.46 3.1 [201]
ZnO NRs 10–23000 110.76 0.1 [1]
5.3.10. Zinc oxide (ZnO) metal oxide based glucose sensors ZnO NRs/prickly 1000–15000 0.097 40 [199]
Electrochemical biosensors can detect bioanalytes using ZnO nano­ ZnO– Cu
ZnO NPs/GCE 1–8.6 631.30 0.043 [12]
structures [204]. Straightforward low-temperature ZnO nanostructure
fabrication results in excellent optical, crystallinity, and electrical
characteristics [2]. ZnO-based hybrid nanostructures with good electron selectivity, sensitivity, miniaturization, biocompatibility, and lifespan,
transport and high surface-to-volume ratios enhanced catalytic activity but they also have certain disadvantages, such as high costs and cali­
in numerous studies [221]. For instance, Xie et al. [222] developed bration needs. The future of non-enzymatic-based sensors appears
glucose sensors based on carbon nanotube nanocomposites and zinc bright, with possible advances in integration, selectivity, calibration,
oxide quantum dots that showed a high sensitivity of 9.36 A M− 1 and a cost, and environmental sustainability leading to more effective and
LOD of 0.208 M. The sensor also delivered repeatable results in 3 s. The accessible glucose monitoring technologies.
sensor also demonstrated good ascorbic acid, sucrose, and dopamine
selectivity over glucose molecules [13,206]. In a different study, Author contributions
Haghparas et al. [75] developed CuO/ZnO microstructures with a high
sensitivity of 1536.80 μA mM− 1 cm− 2 and a wide dynamic range of Kermue Vasco Jarnda, Danqi Wang, & Qurrat-Ul-Ain conceived and
500 nM to 100 mM. drafted the manuscript. Kermue Vasco Jarnda, Danqi Wang, Richmond
Anaman, Varney Edwin Johnson, Qurrat-Ul-Ain, Garmai Prosperity
6. Conclusions and outlook for non-enzymatic glucose sensor Roberts, Pauline Sammumah Johnson, & Bob Wisdom Jallawide Jr
wrote the manuscript and has critically been reviewed by Professor
Significant progress has been made in the development of electro­ Tianhan Kai & Professor Ping Ding. All the authors have approved the
chemical non-enzymatic glucose sensors for the detection of glucose in final version to be submitted for publication.
tears and saliva in recent years. The prospect of continuous glucose
monitoring without an intrusive blood sample has piqued the interest of Funding
these sensors. Recent technological advances have resulted in the cre­
ation of materials with great selectivity for glucose detection, elimi­ This work was supported by the National Natural Science Foundation
nating interference from other compounds found in tears and saliva. of China (grant numbers: 8237363 and 82073607) and the Natural
This selectivity is essential for precise glucose monitoring. New mate­ Science Foundation of Hunan Province (2021JJ30637).
rials have enhanced sensitivity, allowing glucose detection at lower
concentrations. This is especially essential for diabetics who may have
Declaration of Competing Interest
fluctuating glucose levels, and developments in microfabrication pro­
cesses have allowed for the shrinking of these sensors, making them
The authors declare that they have no known competing financial
more comfortable for wearable devices. This increases their capacity for
interests or personal relationships that could have appeared to influence
continuous glucose monitoring. Some new materials have improved
the work reported in this paper.
biocompatibility, lowering the potential for unpleasant reactions when
the sensor comes into contact with tears or saliva for extended periods of
Data Availability
time. Non-enzymatic sensors frequently require calibration to maintain
accuracy, which can be bothersome for users, and fabrication proced­
No data was used for the research described in the article.
ures can be costly, limiting their accessibility to some patients. The
manufacture and disposal of sophisticated materials may have envi­
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Kermue Vasco Jarnda is a PhD candidate at XiangYa School of
Public Health, Central South University in Changsha, China. He
received his master’s degree in public health and preventive
medicine from the same university in 2021. His research fo­
cuses on developing nanomaterials and electrochemical/
colorimetric devices for health inspection and quarantine ap­
plications. Specifically, Mr. Jarnda works on analyte detection
techniques that can advance methods in public health and
preventive medicine. Through his doctoral work, he aims to
apply nanotechnology and biosensing to improve health
inspections.
Professor Ping Ding is affiliated with the Department of
Health Inspection and Quarantine at Xiangya School of Public
Health, Central South University, Changsha, China. As the
corresponding author, her research interests include biomed­
ical functional materials, drug analysis, health-related product
safety detection and evaluation, new technologies and methods
of sanitary inspection, and sensitive biomarkers of environ­
mental media health hazards. Professor Ding has published
influential experimental research on health inspections in
journals such as the Journal of Computational and Theoretical
Nanoscience and the Journal of Pharmacy and Pharmacology.
She serves on the editorial boards of several journals, including
the Chinese Journal of Preventive Medicine and the Interna­
tional Journal of Biological Macromolecules. Professor Ding is an active member of
numerous professional organizations, including the National Health Chemistry Education
Group, the Hunan Inspection and Testing Society, the Hunan Health Economics and In­
formation Society, and the Hunan Preventive Medicine Association. Through her research,
editorial work, and policy engagement, she aims to advance health inspection methods
and improve public health.