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 Page 1 of 26 Materials Chemistry Frontiers

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DOI: 10.1039/D1QM00926E

Poly(3,4-ethylenedioxythiophene) Bearing

Materials Chemistry Frontiers Accepted Manuscript

Fluoro-containing Phenylboronic Acid for
Specific Recognition of Glucose

Wenji Bao1, # Wenfeng Hai1, #, *, Layue Bao1, Fan Yang1, Yushuang Liu1, * Tatsuro Goda2

Jinghai Liu1*
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1. Inner Mongolia Key Laboratory of Carbon Nanomaterials, Nano Innovation

Institute (NII), College of Chemistry and Chemical Engineering, Inner Mongolia

University for Nationalities, Tongliao 028000, China

2. Department of Biomedical Engineering, Faculty of Science and Engineering, Toyo

University, 2100 Kujirai, Kawagoe, 350-8585 Saitama, Japan

*Corresponding author:

hai_wenfeng@163.com (W.F.H.)

yushuangliu1989@163.com (Y.S.L)

jhliu2008@sinano.ac.cn (J.H.L.)

#these authors contribute equally to this paper

Keyword: Electrochemical Biosensor, EDOT-FPBA Monomer, Electropolymerized

Poly(EDOT-FPBA) Film, Glucose Specific Recognition, Physiological Conditions

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DOI: 10.1039/D1QM00926E
Abstract

In this study, we synthesized a new monomer, namely, 3,4-ethylenedioxythiophene

Materials Chemistry Frontiers Accepted Manuscript

bearing a fluoro-containing phenylboronic acid (EDOT-FPBA) for enzyme-free glucose

sensing at physiological conditions. The EDOT-FPBA monomer is electropolymerized

on a glassy carbon electrode (poly(EDOT-FPBA)/GCE) at the constant voltage. The

electrochemical polymerization was optimized to obtain a low impedance film

interface and then characterized by scanning electron microscopy, atomic force

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microscopy, water contact angle measurements, and electrochemical impedance

spectroscopy. The specific interaction and pH dependence between poly(EDOT-

FPBA)/GCE and glucose are detected by electrochemical impedance spectroscopy in a

detection range from 0.05 to 25 mM with a detection limit of 0.05 mM at pH 7.0. The

poly(EDOT-FPBA) exhibits 6 times higher sensitivity under physiological pH conditions

in comparison with phenylboronic acid and 3-pyridylboronic acid conjugated poly(3,4-

ethylenedioxythiophene), due to the introduction of an electron-withdrawing F

substituent into the benzene ring of phenylboronic acid (PBA) reducing the pKa value

of PBA. The sensitivity and the dynamic range cover all levels of blood glucose in the

serum patients with diabetes. Furthermore, with advantages of high sensitivity and

wide detection range, reusability, stability, and mass productivity, the poly(EDOT-FPBA)

as a glucose sensor has a great potential for enzyme-free glucose continuous

monitoring applications.

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DOI: 10.1039/D1QM00926E
1. Introduction

Diabetes is a common chronic non-communicable disease, which has become a

Materials Chemistry Frontiers Accepted Manuscript

serious worldwide public health concern1. Because continuous monitoring of blood

glucose is the key to better control of blood glucose concentration in the treatment of

diabetes, we urgently need new electrode materials and sensor design to meet the

requirements of continuous monitoring of blood glucose. In recent years, the

application of conducting polymers in this field is increasing, among which poly (3.4-
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ethylenedioxythiophene) (PEDOT) and its derivatives have attracted more and more

attention2-3. PEDOT is a typical thiophene conducting polymer, which has good

conductivity, environmental stability in doping state, low redox potential, good film-

forming property, flexibility, and other characteristics4-5. Besides, PEDOT is easy to be

functionalized to achieve specific excitation, detection, and sensing and has great

potential applications in bioelectronics and biosensing 6-7. Because the 3 and 4

positions of 3,4-ethylenedioxythiophene (EDOT) are both replaced by alkoxyl groups,

the alkoxyl substituents reduce the oxidation potential of thiophene monomer and

polymer, making it easier to be electropolymerized and more stable in the redox

(doping and de-doping) cycles8. A series of progress in the perception of the current

of the cerebral cortex and the skin has been made, owing to the nature of good

conductivity, non-toxic, soft, large surface area, and low impedance at

tissue/electrode interfaces9-10, however, PEDOT does not have specific recognition

functions. It is necessary to impart specific recognition elements to PEDOT in advance

for biosensing applications. Two approaches have been explored to introduce specific

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recognition elements to PEDOT. Physical adsorption through mechanical mixing and

doping imparts specific recognition elements into PEDOT. But, the doped small

Materials Chemistry Frontiers Accepted Manuscript

molecules are easy to be de-doped during operation. EDOT functionalization through

chemical synthesis has the advantages of precise modulation of a versatile functional

unit at the molecular scale and adjustable modification density11-12. EDOT derivatives

carrying target-capturing elements have been developed to detect influenza virus13,

DNA14, protein15, sialic acid16, and glucose17.

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Glucose oxidase (GOD) catalyzes the oxidation of beta-D-glucose to gluconic acid

with simultaneous production of hydrogen peroxide. GOD is the most commonly used

glucose recognition element and is widely used in various glucose sensors2, 18. Three

generations of GOD-based biosensors have been proposed, where the first generation

is based on the measurement of hydrogen peroxide formation and oxygen

consumption, the second one uses additional media to transfer electrons from the

enzyme active site to the electrode, and the third one is the direct electron transfer

strategy between GOD and electrode without medium19. Recently, GOD has been

combined with nanomaterials to increase the electron transfer rate, such as noble

metal and transition metal nanoparticles20, nanostructured metal oxides or metal

sulfides21-22, conductive polymers23, carbon nanotubes24 and graphene25. Although

enzymatic glucose sensors commonly provide high selectivity and quantification

platform, these approaches suffering a difficulty of long-term use and storage caused

by denaturing of protein GOD19. This fact has become a driving force for the research

on non-enzymatic synthetic glucose sensors as an alternative strategy for enzymatic

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glucose sensors.

The non-enzymatic glucose sensing approaches can be classified into the artificial

Materials Chemistry Frontiers Accepted Manuscript

(GOD-free) electrochemical oxidation of glucose, and the synthetic ligands with

glucose, e.g., α-cyclodextrin (α-CD) and phenylboronic acid (PBA) derivatives26. The

PBA molecule can reversibly form boronic esters with hydroxyl groups of glucose27-28.

And, the effective coordination binding of PBA with polyol compounds (such as glucose)

requires the sp3 hybridization of boron (B) atom (tetrahedral anion, -B(OH)3-), which
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can only be achieved at a high pH value close to the original pKa (about 8-9) of PBA29-

31. As a result, a limited proportion of sp3 hybridized PBA can bind effectively with

glucose at pH 7.0. Therefore, PBA has poor sensitivity to glucose under physiological

pH conditions, which limits its use for the sensor under physiological pH conditions.

Here, we design a new monomer, namely, 3,4-ethylenedioxythiophene bearing a

fluoro-containing phenylboronic acid (EDOT-FPBA). The electron-withdrawing F

substituent into the benzene ring of phenylboronic acid (PBA) reduces the pKa value

of PBA, endowing the poly(EDOT-FPBA) more sensitivity for glucose under

physiological pH conditions. The optimum electrochemical polymerization conditions,

including solvent, time, and temperature, were explored to obtain a low impedance

film interface for glassy carbon electrode (poly(EDOT-FPBA)/GCE). Compared to the

poly(EDOT-PBA) and poly(EDOT-PyBA), this poly (EDOT-FPBA) has a higher sensitivity

to glucose at pH 7.0, even in the presence of galactose, mannose, dopamine, urea, Na+,

and K+, and a large detection range from 0.05 to 25 mM (R2 = 0.9826) with LOD of 0.05

mM. Practically, the poly(EDOT-FPBA) with high sensitivity and wide detection range

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has successfully utilized in glucose continuous detection in human serum DOI: 10.1039/D1QM00926E
samples,

demonstrating the future potential for enzyme-free glucose monitoring applications.

Materials Chemistry Frontiers Accepted Manuscript

2. Results and Discussion

2.1. Monomer synthesis and optimization of electropolymerization conditions

In this study, we developed a new monomer, namely, EDOT possessing fluorine (F)

containing PBA moiety (EDOT-FPBA), and the electrochemical polymerization was

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carried out on the glassy carbon electrode(GCE) in presence of tetrabutylammonium

perchloride into poly (EDOT-FPBA) for glucose detection under the physiological pH

environment. F was introduced into the benzene ring of PBA as an electron-

withdrawing group, intending to lower the pKa of PBA (Figure 1). The EDOT-FPBA was

successfully synthesized and examined by 1H-NMR and high-resolution ESI-MS (Figure

S1 and S2). The oxidation current of EDOT-FPBA gradually reached an equilibrium and

the capacitance of poly (EDOT-FPBA)/GCE increased when the constant voltage of 1.5

V (versus Ag/AgCl reference electrode) was applied to the GCE working electrode for

10 seconds in the dichloromethane solution containing 10 mM EDOT-FPBA and 100

mM ammonium perchlorate (Figure S3a). Meanwhile, the impedance of poly(EDOT-

FPBA)/GCE increased in contrast to the bare GCE and PEDOT/GCE (Figure S3b),

indicating the poly(EDOT-FPBA) is successfully electropolymerized on GCE. The optimal

electropolymerization conditions were determined by the EIS Nyquist curves with the

lowest charge transfer resistance (Rct) at various monomer solvents (Figure 2a),

electropolymerization time (Figure 2b), and temperature (Figure 2c) for poly(EDOT-

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DOI: 10.1039/D1QM00926E
FPBA)/GCE. The Rct in corresponding Randall equivalent circuit and modeling indicates

the EDOT-FPBA should be electropolymerization with monomer solvent of

Materials Chemistry Frontiers Accepted Manuscript

dichloromethane (CH2Cl2) at 25 °C for 10 s under a constant voltage of 1.5 V (Figure

2d).

2.2. Characterization of poly(EDOT-FPBA) film

The surface morphology of poly(EDOT-FPBA) films on the planar Au electrode was

investigated by scanning electronic microscopy (SEM) and atomic force microscopy

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(AFM). The monomer solvents present significant effects on the morphology, where a

smooth superimposed sheet-like surface with cavity forms in CH2Cl2 containing 100

mM butylammonium perchlorate (Figure 3a), relative to a rough cavity surface in an

aqueous solution containing 100 mM sodium perchlorate (Figure 3b) and a uniform

dense surface in acetonitrile solution100 mM butylammonium perchlorate (Figure 3c).

AFM analysis further reveals the surface profile of poly(EDOT-FPBA) films and

corresponding height variations. Various concave-convex cavity about 100 nm, 80 nm,

80 nm depth was formed with a film thickness of 0.46 μm, 0.25 μm, 0.062 μm prepared

in CH2Cl2 (Figure 3d and 3g), H2O (Figure 3e and 3h), CH3CN (Figure 3f and 3i)

respectively. The XPS analysis demonstrates the existence of F, B, and N elements in

poly(EDOT-FPBA) film, in comparison to the only C, O, and S at PEDOT film (Figure S4),

with the 2.95 a.t.% F, 6.00 a.t.% B, and 5.52% N (Table S1).

2.3. Selectivity and pH dependence of poly(EDOT-FPBA) for glucose recognition

We then examined the selectivity and pH dependence of poly(EDOT-FPBA)/GCE for

glucose recognition by electrochemical impendence spectroscopy (EIS) method at pH

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7.0 in PBS buffer containing a redox couple of 5 mM potassium ferricyanide/potassium

ferrocyanide (K3Fe(CN)6/K4Fe(CN)6). The charge transfer resistance change (ΔRct) of

Materials Chemistry Frontiers Accepted Manuscript

poly(EDOT-FPBA)/GCE denotes the specific interaction towards glucose, galactose,

mannose, dopamine, uric acid, Na+ and K+(5 mM). As shown in Figure 4a, the response

of ΔRct was significantly increased to 53.8 Ω cm2 after the addition of glucose in PBS

buffer, 12 times larger than the one for galactose, 14 times for mannose, 8 times for

dopamine, 8 times for urea, 22 times for Na+ and 10 times for K+, indicating the specific
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recognization towards the glucose. In contrast to the bare GCE and PEDOT/GCE, the

specific glucose recognition originates from the functional FPBA unit conjugated to

PEDOT side chain (Figure 4a insert). Figure 5 shows the possible structure bis-boronate

ester of glucose with poly(EDOT-FPBA). The work of Shinkai et al. shows that glucose

can form a bis-boronate complex with boronic acid derivatives32. In D-glucose the 2-

and 4-hydroxyl groups adopt the same configuration, they both bind to boronate,

while, in D-mannose and D-galactose, the hydroxyl groups adopt the opposing

configurations, this would lead to spatial distortions and prevent the formation of D-

mannose and D-galactose bis-boronate complexes32-33. Thus, the impact of those non-

glucose sugars on glucose detection was minimal.

The pH-dependent glucose-binding ability of poly(EDOT-FPBA) was studied by the

normalized EIS modeling signal (ΔRct/Rct0) for the poly(EDOT-FPBA)/GCE, poly(EDOT-

PBA)/GCE, and poly(EDOT-PyBA). As for the addition of 5 mM glucose at pH 5.0-8.0,

the ΔRct/Rct0 of poly(EDOT-FPBA) at pH 7.0 is 6 times larger than that of poly(EDOT-

PBA), together with almost no signal response for poly(EDOT-PyBA) (Figure 4b). These

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results indicate the electron-withdrawing substituent F at the benzeneDOI:

ring10.1039/D1QM00926E
of

poly(EDOT-FPBA) reducing the electron density on the boron atom and then the pKa

Materials Chemistry Frontiers Accepted Manuscript

of PBA 26, making poly(EDOT-FPBA) can be used for glucose sensing under physiological

pH conditions. In comparison, the introduction of electron-denoting N increases the

pKa of PBA leads to the poly(EDOT-PyBA) showing no more interaction with glucose at

pH 5.0-8.0.

2.4. Electrochemical Glucose sensing using poly(EDOT-FPBA)/GCE

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We further evaluated the sensitivity of poly(EDOT-FPBA)/GCE for glucose sensing

under the EIS method in PBS buffer containing 5 mM a redox couple of 5 mM

K3Fe(CN)6/K4Fe(CN)6 at pH 7.0. The Nyquist curves in Figure 5a show that the

impedance of poly(EDOT-FPBA)/GCE gradually becomes 15 times larger as the

increasing addition of glucose from 0.05 mM to 25 mM. With the Randle equivalent

circuit for modeling and quantitative analysis in Figure 5b, the ΔRct of poly(EDOT-

FPBA)/GCE presents a linear relation with the log10[glucose concentration] in the linear

range of 0.05 to 25 mM (R2=0.9826). And, the detection of limit (LOD) is 0.05 mM. We

also compared the sensing performances in the chemical environment (pH), selectivity,

and linear range with that of literature based on the PBA derivatives as glucose

recognition elements (Table S2). The dynamic range of our system covers not only the

normal human serum glucose level of 3.3−6.1 mM 34, but also can be used to measure

blood glucose in diabetic patients. As we know, the diagnostic criteria for type 2

diabetes are that the random blood glucose level is higher than 11.1 mM35, and the

detection ranges reported in most literature are out of the range of this blood glucose

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level. In addition, our electrode material does not contain heavy metals, andDOI: 10.1039/D1QM00926E
there is

no risk of leaking toxic compounds.

Materials Chemistry Frontiers Accepted Manuscript

Then, we examined the reproducibility of poly(EDOT-FPBA)/GCE for continuous

glucose monitoring over three repeated cycles at 0 and 5 mM. As shown in Figure 6a,

these EIS signals from glucose recognition match very well and are reproducible in the

subsequent three cycles for 5 mM glucose concentration, indicating the capability of

the poly(EDOT-FPBA)/GCE platform for reusable and continuous monitoring. The

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stability for continuous glucose monitoring was also investigated, where the

poly(EDOT-FPBA)/GCE presents a stable ΔRct signal in almost 36 hours in the

subsequent 15 times cycles, with only a little decline in ΔRct signal at the first 5 hours.

2.5. Blood glucose detection by poly(EDOT-FPBA)/GCE

To evaluate the practical application of poly(EDOT-FPBA) electrochemical glucose

sensors in blood glucose detection, we analyzed the glucose concentrations in human

serum. The Nyquist curves in Figure S5 show the increase of Rct as the glucose

concentration rising from 2.5 mM to 10 mM. Through the linear relationship

established for the poly(EDOT-FPBA)/GCE glucose sensor, the recovery of the spiked

glucose (2.5 mM, 5.0 mM, 10 mM) ranges from 96 to 103.2%, calculated based on the

results of three repeated tests in 100 fold diluted human serum sample (Table 1).

These results strongly support the future application of poly(EDOT-FPBA) as an

enzyme-free electrochemical sensor for human blood glucose monitoring. In the

future, we will try to introduce an antifouling-PEDOT to sense whole blood glucose

levels without dilution36.

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3. Conclusion

Glucose sensing functional unit, fluoro-containing phenylboronic acid (FPBA), grafted

Materials Chemistry Frontiers Accepted Manuscript

poly(3,4-ethylenedioxythiophene) was developed for enzyme-free glucose sensing at

physiological pH conditions. The optimum electropolymerized poly(EDOT-FPBA)/GCE

exhibits high sensitivity, wide linear range, and reproducible stability under the EIS

method for glucose detection. Furthermore, the successful application demonstration

for human blood glucose detection directs its future healthcare application as daily
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continuous blood glucose monitoring. This work provides a new approach for the

design of a biocompatible PEDOT-based enzyme-free electrochemical sensor for blood

glucose monitoring at physiological pH conditions.

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4. Materials and methods

4.1 Materials

Materials Chemistry Frontiers Accepted Manuscript

3,4-Dimethoxythiophene, 4-carboxy-3-flurobenzeneboronic acid, 4-(4,6-

Dimethoxy-1,3,5-triazine-2-yl)-4-methlmorpholinium chloride (DMT-MM), 3-chloro-

1,2-propanediol, tetrabutylammonium perchlorate, and dopamine hydrochloride

were purchased from Energy chemical (Shanghai, China). The glucose, mannose,

galactose, and uric acid were purchased from Adamas Reagent, (Shanghai, China).
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Dulbecco’s phosphate-buffered saline (DPBS) was purchased from Solarbio life

sciences (Beijing Solarbio Science & Technology Co. Beijing, China). All reagents were

analytical grade and used without further purifications.

4.2 Instrumentations

1H NMR spectra were collected on Bruker NMR spectrometer (500 MHz Bruker

Corporation, Karlsruhe, Germany). 1H spectra were recorded in D2O. High-resolution

mass spectrometry was recorded on Waters xevo G2-S spectrometer (Waters

Corporation, Milford Massachusetts, USA). Electro-polymerization of EDOT derivatives

and electrochemical measurements were performed using a multi-channel

potentiostat (VMP3, Bio-Logic Science Instruments, France) via EC-Lab software

version 11.10 (bio-logic, France). The Surface morphology of electrodeposited films

was examined by scan electronic microscopy (SEM) (S-4800, Hitachi, Japan) and atomic

force microscopy (AFM) (Asylum Research MFP-3D Origin AFM, Oxford Instruments,

UK) respectively. The AFM probe was using a silicon cantilever with a tip radius of 7

nm (Asylum Research Probes, 6310 Hollister Ave. Santa Barbara, CA 93117). X-ray

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Photoelectron Spectroscopy (XPS) (ESCALAB Xi+, Thermo Fisher Scientific, USA) was

used to measure the elemental compositions.

Materials Chemistry Frontiers Accepted Manuscript

4.3 Synthesis of EDOT-FPBA

The synthesis procedures of amine-functionalized 3,4-ethylenedioxythiophene

(EDOT-NH2) was described in the previous work13. EDOT-NH2 (0.70 g, 4.45 mmol) and

4-carboxy-3-flurobenzeneboronic acid (0.83 g, 4.45 mmol), DMT-MM (1.35 g, 4.90

mmol) were dissolved in methanol (50 mL) and placed in 100 ml eggplant flask and the
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mixture was continuous stirring 1.5 h at 30 °C. After removing methanol and the

residue was purified by column chromatography using silica gel (200−400 mesh)

eluting with a dichloromethane/methanol (15/1, v/v), to give white solid the product

of (4-(((2,3-dihydrothieno[3,4-b][1,4]dioxin-2-yl)methyl)carbamoyl)-3-

fluorophenyl)boronic acid (EDOT-FPBA) (yield 68.6%). 1H NMR (500 MHz, D2O) δ 7.35

(t, J = 7.5 Hz, 1H), 7.35 (t, J = 7.5 Hz, 1H), 7.22 (d, J = 7.6 Hz, 1H), 7.22 (d, J = 7.6 Hz, 1H),

7.11 (d, J = 12.7 Hz, 1H), 7.11 (d, J = 12.7 Hz, 1H), 6.24 (s, 2H), 6.24 (s, 2H), 4.25 – 4.16

(m, 1H), 4.08 (dd, J = 11.9, 2.1 Hz, 1H), 3.85 (dd, J = 12.0, 6.7 Hz, 1H), 3.49 (d, J = 5.7

Hz, 2H), 3.16 (s, 2H). Theoretical m/z for C14H14BFNO5S+ (M+H+) at 338.07,

experimentally found at 338.26.

4.4 Preparation of poly(EDOT-FPBA) films

A glassy carbon electrode(GCE) of 3 mm diameter (Sanshe Industrial Co. Shanghai,

China) was used as a working electrode. Before electropolymerization of film, bare

GCEs were polished using 0.05 m alumina slurries then washed with ultrapure water.

For the optimized electropolymerization conditions, we have prepared poly(EDOT-

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FPBA) films at different conditions. The first is to choose different solvents toDOI: 10.1039/D1QM00926E
dissolve

EDOT-FPBA monomer, (i) 10 mM EDOT-FPBA monomer and 100mM

Materials Chemistry Frontiers Accepted Manuscript

tetrabutylammonium perchlorate in dichloromethane, and in (ii) acetonitrile, (iii) 10

mM EDOT-FPBA monomer and 100 mM sodium perchlorate in water. Then, take 1 ml

of the mixed solution in a glass cell, and then introduce the three electrodes (a glassy

carbon working electrode, a platinum plate counter electrode, and an Ag/AgCl

electrode) which connected to the potentiostat in the cell. The electropolymerization

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was performed at a constant potential of 1.5 V (vs Ag/AgCl) for 10 seconds at room

temperature. The second is to choose different electropolymerization times. 10 mM

EDOT-FPBA monomer and 100 mM tetrabutylammonium perchlorate were

electropolymerized in dichloromethane at room temperature for 10, 20, 30 seconds at

1.5 V (vs Ag/AgCl) constant potential. The third is to choose different

electropolymerization temperatures. 10 mM EDOT-FPBA monomer and 100 mM

tetrabutylammonium perchlorate were electropolymerized in dichloromethane at 0 °C

and 25 °C for 10 seconds, at 1.5 V (vs Ag/AgCl) constant potential.

4.5 General Characterization

EIS measurements were performed in a solution of 5 mM K3Fe(CN)6/K4Fe(CN)6 +

1×DPBS, under an AC amplitude of 50 mV superimposed on a DC bias of +0.2 V and a

frequency range of 0.1 Hz to 10 kHz. Three electrodes consist of a polythiophene

deposited working electrode, an Ag/AgCl (in 3.3 M KCl) reference electrode, and a

platinum plate counter electrode. The surface observations of the electrodeposited

films were performed by SEM at the accelerating potential of 5.0 kV and a working

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distance of 8 mm. The polythiophene derivative films were prepared on a flat gold-

sputtered silicon electrode by electrodeposition process and treated with a Pt coater

Materials Chemistry Frontiers Accepted Manuscript

for 30 s before SEM observations. The micro-and nano-structures of polythiophene

films were studied by the tapping mode imaging technique AFM with the following

parameters: setpoint: 668 mV, drive amplitude:33.5 mV, integral gain:5.47. The

samples were prepared by electrodeposition of polymers onto a gold electrode. XPS

spectrometer is equipped with the Al Kα radiation source (1486.6 eV). The takeoff
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angle of the photoelectron analyzer was 90°.

4.6 Glucose sensing in PBS

The glucose in the range from 0.05 to 25 mM in 1×DPBS(pH 7.0) was measured

by EIS in the presence of 5 mM K3Fe(CN)6/K4Fe(CN)6 at room temperature. A multi-

channel potentiostat with a three electrodes cell was used. The glassy carbon

electrode deposited by polythiophene was used as the working electrode, platinum

plate as the counter electrode, and an Ag / AgCl (in 3.3 M KCl) as the reference

electrode. Rct is determined by fitting Randle’s equivalent circuit to the Nyquist

diagram. The mean and standard deviation were obtained from three independent

tests.

4.7 Glucose detection in serum sample

Blood samples were provided by the Affiliated Hospital of Inner Mongolia University

after the approval of the school ethics committee. Blood samples were pretreated

before use. The samples were centrifuged at 5000 rpm for 20 minutes, then filtered

with 0.22 membrane, and the supernatant was used for the subsequent tests. In the

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EIS recovery test, the collected serum were diluted 100 times using PBS buffer, and

then glucose was added to obtain the final concentration. The recoveries were

Materials Chemistry Frontiers Accepted Manuscript

calculated from three concentrations of glucose spiked human serum samples.
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5. Figures and tables

Materials Chemistry Frontiers Accepted Manuscript

Figure 1. Scheme for the synthetic route of an EDOT derivative possessing a 3-
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fluorophenylboronic acid group (EDOT-FPBA) and electrochemical polymerization of

EDOT-FPBA into (poly(EDOT-FPBA)) on glassy carbon electrode(GCE) for glucose

detection.

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DOI: 10.1039/D1QM00926E

(a) (b)
1500 Electropolymerization solvents 1500 Electropolymerizaition time

Materials Chemistry Frontiers Accepted Manuscript

CH2Cl2 10 s
H2O 20 s
1000 1000
CH3CN

Z''(Ω)
Z''(Ω)

30 s

500 500

0 0
0 500 1000 1500 2000 2500 3000 0 500 1000 1500 2000

(c) Z'(Ω) (d) Z'(Ω)

1500 Electropolymerization temperature Q

CH3CN
200
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0℃
Rct (Wcm2)
25℃ Rsol
1000 150
Z''(Ω)

Rct

H2 O
100

CH2Cl2
500
50

0 0
0 500 1000 1500 2000 10 2030 0 25
Z'(Ω) Solvents Time(s) Temperature (°C )

Figure 2. Optimization of electropolymerization conditions of poly(EDOT-FPBA)/GCE

examined by EIS Nyquist curves in PBS buffer containing 5 mM potassium

ferricyanide/potassium ferrocyanide (K3Fe(CN)6/K4Fe(CN)6) redox pairs.

Electropolymerization at a constant voltage of 1.5 Vunder (a) solvents,(b) times. and

(c) temperatures. (d) Charge transfer resistance (Rct) for the poly(EDOT-FPBA)/GCE at

different conditions. Inset: corresponding equivalent circuit and modeling. . The data

are shown as the mean ± SD (n = 3).

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DOI: 10.1039/D1QM00926E

(a) (b) (c)

Materials Chemistry Frontiers Accepted Manuscript

CH2Cl2 H2O CH3CN

1 μm 1 μm 1 μm

10 μm 10 μm 10 μm

(d) (e) (f)

CH2Cl2 H2O CH3CN
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(g) (h) (i)

-0.9 CH Cl -0.5 0.3
2 2
H2O CH3CN
-0.6
-1.0 -0.7 0.2
Height(m)
Height(m)

Height(m)
-1.1 -0.8
-1.2 -0.9 0.1
0.46 m -1.0
-1.3 -1.1 0.0
-1.4 -1.2 0.25 m 0.062 m
-1.3 -0.1
-1.5
0 20 40 60 80 100 120 0 20 40 60 80 100 120 0 20 40 60 80 100 120
Scan length(m) Scan length(m) Scan length(m)

Figure 3. Morphology of poly(EDOT-FPBA) electropolymerized on planar Au electrode.

SEM images for the monomer solvents of (a) CH2Cl2, (b) H2O, (c) CH3CN.. (d-f) AFM

scanning profile and (g-i) Height profile.

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DOI: 10.1039/D1QM00926E

(a) (b)
60 poly(EDOT-FPBA) 60
2.0 poly(EDOT-FPBA)
poly(EDOT-FPBA)

Materials Chemistry Frontiers Accepted Manuscript

DRct(Wcm2)
40
poly(EDOT-PBA)
poly(EDOT-PyBA)
1.5
DRct(Wcm2)

20
40

DRct/R0ct
PEDOT GCE
0
1.0
20
0.5

0 0.0
ce tose os
e ine Na
+ K+ ea 5 6 7 8
co c nn am Ur
Glu Gla Ma Dop
pH

Figure 4. Selectivity and pH dependence of poly(EDOT-FPBA) for glucose recognition

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in PBS buffer (7.0) containing a redox couple of 5 mM K3Fe(CN)6/K4Fe(CN)6. (a) The

charge transfer resistance changes of poly(EDOT-FPBA)/GCE after the addition of 5 mM

glucose, galactose, mannose, dopamine, urea, Na+, and K+. Inset: Glucose (5mM) for

the GCE with or without PEDOT deposited as controls. (b) pH dependence of the

normalized EIS signal (ΔRct/Rct0) for the poly(EDOT-FPBA)/GCE (red curve), poly(EDOT-

PBA)/GCE (blue curve), and poly(EDOT-PyBA)/GCE (green curve) at 5 mM glucose in

PBS buffer. The data are shown as the mean ± SD (n = 3).

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DOI: 10.1039/D1QM00926E

Materials Chemistry Frontiers Accepted Manuscript

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Figure 5. The possible structure of a bis-boronate complex of glucose with

poly(EDOT-FPBA)

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(a) (b) DOI: 10.1039/D1QM00926E
0(mM) 200
2000 poly(EDOT-FPBA 0.05
0.10
160
0.25

DRct (Wcm2)
1500 0.50 y=66.41lg(x)+85.19

Materials Chemistry Frontiers Accepted Manuscript

Z(W)

1.00 120 2
2.50 R =0.9826
1000 5.00
80
10.00
25.00
500 40

0
0
0 1000 2000 3000 4000 0.01 0.1 1 10 100

Z(W) lg[Glucose concentration] (mM)

(c) (d)
1500 0 mM-1 100
0 mM-2
1200 0 mM-3
80
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DRct (Wcm2)
5 mM-1
900 5 mM-2
Z(W)

5 mM-3 60

600
40
300
20
0
0
0 500 1000 1500 2000 2500 3000 0 5 10 15 20 25 30 35 40
Z(W) Time (hour)
Figure 6. Poly(EDOT-FPBA)/GCE for the detection of glucose by EIS method in PBS (pH
7.0) containing a redox couple of 5 mM K3Fe(CN)6/K4Fe(CN)6. (a) Nyquist curves for
glucose concentrations from 0.05 to 25 mM. (b) Linear relations of ΔRct and
lg[Glucose concentration], R2=0.9826. The data are shown as the mean ± SD (n = 3).
Inset: Randle equivalent circuit and modeling. (c) EIS Nyquist curves showing
continuous monitoring of glucose in three repeated cycles at 0 and 5.0 mM glucose
concentration. (d) Corresponding ΔRct signal after storing for 2, 5, 12, 36 hours.

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DOI: 10.1039/D1QM00926E

Table 1. Detection of glucose in human serum using the poly(EDOT-FPBA)/GCE

Materials Chemistry Frontiers Accepted Manuscript

Test No. Add (mM) Found (mM) Recovery (%)

1 2.5 2.58 103.2

2 5.0 4.82 96

3 10.0 9.73 97.3

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DOI: 10.1039/D1QM00926E
References
(1) Williams, R.; Karuranga, S.; Malanda, B.; Saeedi, P.; Basit, A.; Besancon, S.; Bommer, C.; Esteghamati,
A.; Ogurtsova, K.; Zhang, P.; Colagiuri, S. Global and regional estimates and projections of diabetes-

Materials Chemistry Frontiers Accepted Manuscript

related health expenditure: Results from the International Diabetes Federation Diabetes Atlas, 9th
edition. Diabetes Res. Clin. Pract. 2020, 162, 108072.
(2) Sehit, E.; Altintas, Z. Significance of nanomaterials in electrochemical glucose sensors: An updated
review (2016-2020). Biosens. Bioelectron. 2020, 159, 112165.
(3) Wei, M.; Qiao, Y.; Zhao, H.; Liang, J.; Li, T.; Luo, Y.; Lu, S.; Shi, X.; Lu, W.; Sun, X. Electrochemical non-
enzymatic glucose sensors: recent progress and perspectives. Chem. Commun. (Camb) 2020, 56, 14553-
14569.
(4) Donahue, M. J.; Sanchez-Sanchez, A.; Inal, S.; Qu, J.; Owens, R. M.; Mecerreyes, D.; Malliaras, G. G.;
Martin, D. C. Tailoring PEDOT properties for applications in bioelectronics. Mat. Sci. Eng. R. 2020, 140,
100546.
Published on 23 August 2021. Downloaded on 9/1/2021 12:04:47 AM.

(5) Mantione, D.; Del Agua, I.; Sanchez-Sanchez, A.; Mecerreyes, D. Poly(3,4-ethylenedioxythiophene)
(PEDOT) Derivatives: Innovative Conductive Polymers for Bioelectronics. Polymers (Basel) 2017, 9, 354.
(6) Hui, Y.; Bian, C.; Xia, S. H.; Tong, J. H.; Wang, J. F. Synthesis and electrochemical sensing application
of poly(3,4-ethylenedioxythiophene)-based materials: A review. Anal. Chim. Acta 2018, 1022, 1-19.
(7) Gueye, M. N.; Carella, A.; Faure-Vincent, J.; Demadrille, R.; Simonato, J. P. Progress in understanding
structure and transport properties of PEDOT-based materials: A critical review. Prog. Mater. Sci. 2020,
108, 100616.
(8) Ko, H. C.; Park, S.-a.; Lee, H. Characteristics of dual-type electrochromic device based on poly(3-
tetradecylthiophene) and poly(3,4-ethylenedioxythiophene). Synth. Met. 2004, 143, 31-35.
(9) Parashar, K.; Prajapati, D.; McIntyre, R.; Kandasubramanian, B. Advancements in Biological Neural
Interfaces Using Conducting Polymers: A Review. J. Ind. Eng. Chem. 2020, 59, 9707-9718.
(10) Araki, T.; Bongartz, L. M.; Kaiju, T.; Takemoto, A.; Tsuruta, S.; Uemura, T.; Sekitani, T. Flexible neural
interfaces for brain implants-the pursuit of thinness and high density. Flexible and Printed Electronics
2020, 5, 043002.
(11) Balint, R.; Cassidy, N. J.; Cartmell, S. H. Conductive polymers: Towards a smart biomaterial for tissue
engineering. Acta Biomater. 2014, 10, 2341-2353.
(12) Jarosz, T.; Stolarczyk, A.; Glosz, C. Recent Advances in the Electrochemical Synthesis of Copolymers
Bearing pi-Conjugated Systems and Methods for the Identification of their Structure. Curr. Org. Chem.
2020, 24, 339-353.
(13) Hai, W. F.; Goda, T.; Takeuchi, H.; Yamaoka, S.; Horiguchi, Y.; Matsumoto, A.; Miyahara, Y. Specific
recognition of human influenza virus with PEDOT bearing salic acid-terminated trisaccharides. ACS Appl.
Mater. Interfaces 2017, 9, 14162-14170.
(14) Galan, T.; Prieto-Simon, B.; Alvira, M.; Eritja, R.; Gotz, G.; Bauerle, P.; Samitier, J. Label-free
electrochemical DNA sensor using "click"-functionalized PEDOT electrodes. Biosens. Bioelectron. 2015,
74, 751-756.
(15) Goda, T.; Toya, M.; Matsumoto, A.; Miyahara, Y. Poly(3,4-ethylenedioxythiophene) Bearing
Phosphorylcholine Groups for Metal-Free, Antibody-Free, and Low-Impedance Biosensors Specific for
C-Reactive Protein. ACS Appl. Mater. Interfaces 2015, 7, 27440-27448.
(16) Hai, W. F.; Pu, S. H.; Wang, X.; Bao, L. M.; Han, N.; Duan, L. M.; Liu, J. H.; Goda, T.; Wu, W. M. Poly(3,4-
ethylenedioxythiophene) Bearing Pyridylboronic Acid Group for Specific Recognition of Sialic Acid.

24
 Page 25 of 26 Materials Chemistry Frontiers

View Article Online

Langmuir 2020, 36, 546-553. DOI: 10.1039/D1QM00926E
(17) Huang, P.-C.; Shen, M.-Y.; Yu, H.-h.; Wei, S.-C.; Luo, S.-C. Surface Engineering of Phenylboronic Acid-
Functionalized Poly(3,4-ethylenedioxythiophene) for Fast Responsive and Sensitive Glucose Monitoring.
ACS Appl. Bio Mater. 2018, 1, 160-167.

Materials Chemistry Frontiers Accepted Manuscript

(18) Hwang, D. W.; Lee, S.; Seo, M.; Chung, T. D. Recent advances in electrochemical non-enzymatic
glucose sensors - A review. Anal. Chim. Acta 2018, 1033, 1-34.
(19) Witkowska Nery, E.; Kundys, M.; Jelen, P. S.; Jonsson-Niedziolka, M. Electrochemical Glucose
Sensing: Is There Still Room for Improvement? Anal. Chem. 2016, 88, 11271-11282.
(20) Fang, L.; Liu, B.; Liu, L.; Li, Y.; Huang, K.; Zhang, Q. Direct electrochemistry of glucose oxidase
immobilized on Au nanoparticles-functionalized 3D hierarchically ZnO nanostructures and its
application to bioelectrochemical glucose sensor. Sens. Actuators B Chem. 2016, 222, 1096-1102.
(21) Song, Y.; Xu, M.; Gong, C.; Shen, Y.; Wang, L.; Xie, Y.; Wang, L. Ratiometric electrochemical glucose
biosensor based on GOD/AuNPs/Cu-BTC MOFs/macroporous carbon integrated electrode. Sens.
Actuators B Chem. 2018, 257, 792-799.
Published on 23 August 2021. Downloaded on 9/1/2021 12:04:47 AM.

(22) Jeong, J.-M.; Yang, M.; Kim, D. S.; Lee, T. J.; Choi, B. G.; Kim, D. H. High performance electrochemical
glucose sensor based on three-dimensional MoS2/graphene aerogel. J. Colloid Interface Sci. 2017, 506,
379-385.
(23) Al-Sagur, H.; Komathi, S.; Khan, M. A.; Gurek, A. G.; Hassan, A. A novel glucose sensor using lutetium
phthalocyanine as redox mediator in reduced graphene oxide conducting polymer multifunctional
hydrogel. Biosens. Bioelectron. 2017, 92, 638-645.
(24) Comba, F. N.; Romero, M. R.; Garay, F. S.; Baruzzi, A. M. Mucin and carbon nanotube-based
biosensor for detection of glucose in human plasma. Anal. Biochem. 2018, 550, 34-40.
(25) Pu, Z.; Wang, R.; Wu, J.; Yu, H.; Xu, K.; Li, D. A flexible electrochemical glucose sensor with composite
nanostructured surface of the working electrode. Sens. Actuators B Chem. 2016, 230, 801-809.
(26) Matsumoto, A.; Matsumoto, H.; Maeda, Y.; Miyahara, Y. Simple and robust strategy for
potentiometric detection of glucose using fluorinated phenylboronic acid self-assembled monolayer.
Biochim. Biophys. Acta Gen. Subj. 2013, 1830, 4359-4364.
(27) Matsumoto, A.; Miyahara, Y. 'Borono-lectin' based engineering as a versatile platform for
biomedical applications. Sci. Technol. Adv. Mater. 2018, 19, 18-30.
(28) Zhai, W.; Sun, X.; James, T. D.; Fossey, J. S. Boronic Acid-Based Carbohydrate Sensing. Chem. Asian.
J. 2015, 10, 1836-1848.
(29) Sun, X.; James, T. D. Glucose Sensing in Supramolecular Chemistry. Chem. Rev. 2015, 115, 8001-
8037.
(30) Nishiyabu, R.; Kubo, Y.; James, T. D.; Fossey, J. S. Boronic acid building blocks: tools for sensing and
separation. Chem. Commun. (Camb) 2011, 47, 1106-1123.
(31) Zhang, Y. J.; Guan, Y.; Zhou, S. Q. Synthesis and volume phase transitions of glucose-sensitive
microgels. Biomacromolecules 2006, 7, 3196-3201.
(32) Shiomi, Y.; Saisho, M.; Tsukagoshi, K.; Shinkai, S. Specific complexation of glucose with a
diphenylmethane-3,3′-diboronic acid derivative: correlation between the absolute configuration of
mono- and di-saccharides and the circular dichroic activity of the complex. J. Chem. Soc. Perkin 1 1993,
2111-2117.
(33) Takeuchi, M.; Mizuno, T.; Shinkai, S.; Shirakami, S.; Itoh, T. Chirality sensing of saccharides using a
boronic acid-appended chiral ferrocene derivative. Tetrahedron Asymmetry 2000, 11, 3311-3322.
(34) Martins, P. J.; Haas, M.; Obici, S. Central nervous system delivery of the antipsychotic olanzapine

25
 Materials Chemistry Frontiers Page 26 of 26

View Article Online

induces hepatic insulin resistance. Diabetes 2010, 59, 2418-2425. DOI: 10.1039/D1QM00926E
(35) de Vegt, F.; Dekker, J. M.; Ruhé, H. G.; Stehouwer, C. D. A.; Nijpels, G.; Bouter, L. M.; Heine, R. J.
Hyperglycaemia is associated with all-cause and cardiovascular mortality in the Hoorn population: the
Hoorn Study. Diabetologia 1999, 42, 926-931.

Materials Chemistry Frontiers Accepted Manuscript

(36) Goda, T.; Miyahara, Y. Electrodeposition of Zwitterionic PEDOT Films for Conducting and Antifouling
Surfaces. Langmuir 2019, 35, 1126-1133.
Published on 23 August 2021. Downloaded on 9/1/2021 12:04:47 AM.

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