Materials Letters 309 (2022) 131324

Contents lists available at ScienceDirect

Materials Letters
journal homepage: www.elsevier.com/locate/matlet

Molecularly imprinted polymer-silica nanocomposite based potentiometric

sensor for early prostate cancer detection
S. Fernández-Puig a, A.R. Lazo-Fraga b, Brian A. Korgel c, Goldie Oza a, Ateet Dutt d,
V. Vallejo-Becerra e, A.C. Valdés-González b, A.U. Chávez-Ramírez a, *
a
Centro de Investigación y Desarrollo Tecnológico en Electroquímica S. C. Querétaro, Mexico
b
Instituto de Ciencia y Tecnología de Materiales, Habana, Cuba
c
McKetta Department of Chemical Engineering and Texas Materials Institute, The University of Texas at Austin, Austin, Texas 78712-1062, United States
d
Instituto de Investigaciones en Materiales, UNAM, Ciudad de México, Mexico
e
Universidad Autónoma de Querétaro, Querétaro, Mexico

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

Keywords: A novel nanocomposite based on molecularly imprinted polymer (MIP) polymerized over silica nanoparticles (Si)
Sarcosine is used for the development of an all-solid-state (ASS) potentiometric sensor for determination of sarcosine, a
potentiometric ASS sensor biomarker for Prostate Cancer (PCa). This MIP-Si sensor has shown high selectivity in phosphate-buffered so­
Molecularly imprinted polymer
lution (PBS) and simulated body fluid (SBF). We have obtained a linear response (10− 5–10− 8 mol/L), with a low
detection limit (7.8 × 10− 8 mol/L) and a quick response time close to 30 s, being stable for at least 150 days. It is
corroborated that the sensor is a stable, reproducible, and sensitive biosensing device for PCa detection.

1. Introduction This work proposes development of the first potentiometric minia­

Prostate cancer (PCa) is a type of carcinoma, [1] ranked 2nd in the
world as per World Cancer Research Fund (WCRF). Presently, PCa
detection is done by determining prostate-specific antigen (PSA) in
blood. Increased PSA concentration in blood is due to benign prostatic
hypertrophy and prostatitis. It also shows non-specificity in the 2–10 ng/
L range since there is a 40 % chance of exhibiting false-positive results at
those concentrations [2]. This motivated researchers to identify one
metabolic biomarker known as sarcosine (N-methylglycine) found in
muscles and other body fluids. The concentration of sarcosine in
different body fluids is in the range of (0.9–22.2 μmol/L) [3]. It is known
that the sarcosine concentration increases during the progression of PCa.
Potentiometric sensors are currently used to detect analytes based on
PVC membranes with molecularly imprinted polymers (MIP) as recog­
nition element [4]. MIPs are highly specific and selective polymers that
act as a sorbent to extract the target molecules. They offer mechanical
stability in different solvents and are stable at different pHs, tempera­
tures and elevated pressures. Hence, MIP-based biosensors are efficient
with reduced response time [4]. In this work, silica nanoparticles are
used for the MIP preparation since they render greater stability, higher
permeability, thermostability, and biocompatibility [5].
turized ASS sensor for the recognition of sarcosine. Low limit of detec­
tion (LOD), response time, and linearity were observed and discussed.
2. Experimental
Si nanoparticle synthesis: 90 mL ethanol, 9 mL H2O and 6 mL
triethoxysilane (TEOS) were continuously stirred for 15 min, then 4 mL
of ammonia (NH3) was added and allowed to stir for 1 h. Then, Si
nanoparticles were centrifuged and washed with ethanol [5].
MIP-Si synthesis: 290 mg of Si nanoparticles were added to a
mixture of 0.5 mmol of sarcosine, functional monomer methacrylic acid
(MAA), and 4 mL of acetonitrile. The crosslinker (EGDMA) and the
radical initiator (AIBN) were then added. The mixture was degasified
and ultrasonicated for 15 min, placed in a thermal bath, and incubated
at 60 ◦ C. Non-Imprinted Polymers (NIP) were also prepared without
template molecules [6].
Potentiometric ASS sensor: The sensor was assembled using the
conductive material mixture, packed into micropipette tips and selective
membranes containing the MIP was deposited on top of the tip for
potentiometric-based sarcosine analysis (Fig. 1) [7].

* Corresponding author at: Parque Tecnológico Querétaro s/n, Sanfandila Pedro Escobedo, Querétaro 76703, Mexico.
E-mail addresses: camiloaristides@yahoo.es (A.C. Valdés-González), achavez@cideteq.mx (A.U. Chávez-Ramírez).

https://doi.org/10.1016/j.matlet.2021.131324
Received 11 September 2021; Received in revised form 11 November 2021; Accepted 16 November 2021
Available online 19 November 2021
0167-577X/© 2021 Elsevier B.V. All rights reserved.
S. Fernández-Puig et al. Materials Letters 309 (2022) 131324

Fig. 1. Scheme a) Potentiometric ASS sensor: b) epoxy composite conductor c) liquid membrane d) reference electrode Ag+/AgCl e) micrograph of the liquid
membrane with 24 h activation.

Fig. 2. A) SEM of MIP over Si nanoparticles B) TEM of MIP over Si C) RAMAN spectrum of sarcosine, MIP and NIP and D) FTIR spectra of MIP and NIP.

3. Results and discussion while silica nanoparticles act as structural support for the MIPs.
The Scanning electron microscopic (SEM) image in Fig. 2A exhibits
The important aspect of MIP-Si nanocomposites is the stable nanoaggregates of MIP-Si with high surface area. S1 shows spherical Si
complexation of templates (sarcosine), functional monomers (MAA), nanoparticles in the size range of 100–270 nm. The transmission

2
S. Fernández-Puig et al.
Fig. 3. A) The effect of pH on the ASS potentiometric sensor (conc: 10− 4 mol/L, room temperature (RT)); B) Effect of time of activation 24, 48, 72 h (sarcosine, conc
10− 4 mol/L, pH 7, RT); C) Comparison measurements of ASS sensors; D) Response time of the sensor.
electron microscopic (TEM) image (Fig. 2B) showed the internal texture
changes in the formation of MIP-Si material. The EDS analysis confirmed
the presence of Si in the structure of the polymer (S3). S4 describes the
elements present in MIP-Si after polymerization. It is observed that the
pores possess better texture and the nanoparticles are irregular in shape.
The BET surface area showed higher surface area and porosity of ma­
terials. The BET parameters obtained from the MIP were as follows:
surface area 298.75 m2/g, pore volume 0.72 m3/g and pore radius 10.7
Å and for NIP: surface area 263.83 m2/g, pore volume 0.7 m3/g and pore
radius 10.3 Å. The BET pore volume of around 0.7 cm3/g is optimum for
the increased surface area [8].
RAMAN spectrum for the sarcosine (solid) and polymers (MIP-Si and
NIP-Si) in the region of 1000 to 3200 cm− 1, are shown in Fig. 2C. For
sarcosine, the band 3025 cm− 1 corresponds to the stretching of C–H in
the methyl group, and 2954 cm− 1 corresponds to the stretching of N–H
between the NH2 as well as the carbonyl groups. The signals that appear
at 1384 and 1472 cm− 1 are assigned to the combination of C–C
stretching, with the stretching C–– O, balancing of NH2, and movement
of NH2+ symmetric deformation of the CH3 group. On the other hand,
these signals don’t appear in the MIP-Si and NIP-Si, thus confirming that
no template remains in the MIP after washing [9].
FTIR spectra of MIP-Si and NIP-Si (Fig. 2D) shows 1714 cm− 1 is a
characteristic of the carbonyl group stretching. The interaction between
the template and the monomer shifted peaks in the MIP-Si spectrum,
which showed –CH stretch at 2964–2601 cm− 1. At 1054 cm− 1 median
signal corresponds to the stretching and rocking vibrations of C–N,
C–H–H bands respectively.
Sarcosine has two pKA values and hence possesses three different
charged species depending on the pH. As shown in Fig. 3A the potential
(E) in the sensor increases as pH increases till pH 7 and then starts
decreasing as pH increases to 8. At pH < 4, the group (–NH− ) was
protonated, and the sarcosine did not react with functional groups in the
3
Materials Letters 309 (2022) 131324
membranes. This may be because there is little interaction of sarcosine
with the groups in the membrane confined polymer at these pHs. With
increase in pH (4–7), the sarcosine group reacted with functional groups
in MIP (–COO− ), leading to electrostatic interaction; thus the physio­
logical pH 7 is optimum for all future studies.
As shown in Fig. 3B, increasing time does not improve the capacity of
the sensors since the slope decreases, hence such an extended activation
time of the sites is not necessary. At 24 h the sensor showed better
characteristic if we compare the slope between the sensors- 34.7, 10.5,
and 11.4 mV/Dec at 24, 48 and 72 h respectively. We can conclude that
24 h is the time necessary for membrane activation. The imprinting ef­
fect could be observed by comparing sensor ASS ISE-MIP with ISE-NIP
(S5). The sensor relying on the imprinted material showed much
higher slopes and lower LOD than those with non-imprinted ones
(Fig. 3C).
4. Calibration range
The selectivity of potentiometric sensor ASS is mainly expressed in
potentiometric selectivity coefficients KpotA,B.
aA
KABPot =
aBZA/ZB
When aA is the activity of primary ion and aB is the activity of sec­
ondary ion. Overall the Kpot values ranged from 1.45 × 10− 3 to 3.57 ×
10− 3, showing no interference of the species for sarcosine. The relative
order for interferences was creatinine > uric acid > L-histidine > biotin
> glycine (see S6). The response time was examined by recording the
potential readings at time intervals of 5 s over 1 min. Fig. 3D confirms
the response time of 30 s for the sensor and remained constant. This
response is due to selective interaction between sarcosine and MIP in the
membrane. For lifetime evaluation of the miniaturized sensors (ASS),
the same electrodes were prepared and tested for 6 months, five times
S. Fernández-Puig et al. Materials Letters 309 (2022) 131324

Table 1 González: Conceptualization, Writing – review & editing. A.U. Chávez-

Comparison of existing sarcosine sensors as reported in the literature. Ramírez: Supervising, Writing – review & editing.
Method Support LOD (mol/ pH Time Reference
L) (days)

Enzymatic Sarcosine 1⋅10− 6

7.5 – [12] Declaration of Competing Interest
oxidase (SOX) –
Amplex red The authors declare that they have no known competing financial
8
Potentiometric SOX/EDC/NHS/ 1.6⋅10− 7.2 60 [7] interests or personal relationships that could have appeared to influence
Au/ZnONPs/
SPEs
the work reported in this paper.
7
Amperometric PVA-Ag/AuNPS- 5⋅10− 8 NR [13]
pphEOS-SOD/GE Acknowledgements
− 5
Potentiometric p-AMTa/GCE 5⋅10 7.2 NR [14]
Amperometric Pt@ZIF8/GCE 1.06⋅10− 6 7 3 [15]
Potentiometric Sensor ASS 7.8⋅10− 8 7 150 Present The authors express their gratitude to the UT System-CONACYT
work Grant (2020-76B) for the financial support. Goldie Oza also thanks
Conacyt for the financial support under the Catedras Project 746.

per day. It was found that the electrodes worked well over 150 days
without showing significant difference in slope and response time, but
later the slopes start decreasing due to aging or membrane poisoning
(Fig. S8).
5. Analytical application
The determination of sarcosine was carried out in synthetic urine
samples (Table S7) [10]. The sensor was calibrated under optimum
conditions (pH 7, room temperature, 24 h of activation). Blank synthetic
urine was spiked and analyzed for sarcosine concentration in the range
of 10− 8–10− 5 mol/L and was linear. A good response was found and
showed recoveries ranging 92–94 % (see Table S5). The detection limit
of the sensor (LOD) was 7.8 × 10− 8 mol/L [11]. The sensor presents a
promising application in analyzing physiological samples (9.0 ×
10− 7–2.2 × 10− 5 mol/L). Table 1 represents a comparison of sarcosine
sensors with the sensor developed in this work (Table 1).
6. Conclusions
The characterization of the new MIP synthesized on silica nano­
particles (Si) allowed the development of a all solid-state potentiometric
sensor (MIP-Si-ASS) selective towards the sarcosine molecule and has a
fast response time, a wide linear range and a low detection limit,
properties that demonstrate the analytical applicability of the proposed
sensor for PCa diagnostic applications.
CRediT authorship contribution statement
S. Fernández-Puig: Writing – original draft. A.R. Lazo-Fraga:
Methodology. Brian A. Korgel: Methodology, Evaluation, Supervising.
Goldie Oza: Writing – review & editing. Ateet Dutt: Writing – review &
editing. V. Vallejo-Becerra: Validation, Visualization. A.C. Valdés-
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.matlet.2021.131324.
References
[1] M. Dean, H. Lou, Asian J. Androl. 15 (2013) 309, https://doi.org/10.1038/
aja.2013.29.
[2] A. Heidenreich, J. Bellmunt, M. Bolla, S. Joniau, M. Mason, V. Matveev, N. Mottet,
H. Schmid, T. Van der Kwast, T. Wiegel, Actas Urologicas Espanolas 35 (2011) 501,
https://doi.org/10.1016/j.eururo.2010.10.039.
[3] Y. Jiang, X. Cheng, C. Wang, Y. Ma, Anal. Chem. 82 (2010) 9022, https://doi.org/
10.1021/ac1019914.
[4] J.J. BelBruno, Chem. Rev. 119 (2018), https://doi.org/10.1021/acs.
chemrev.8b00171.
[5] B. Fresco-Cala, A.D. Batista, S. Cárdenas, Molecules 25 (2020), https://doi.org/
10.3390/molecules25204740.
[6] C. Huan, S. Shu-Qing, Chin. Phys. B 23 (2014), https://doi.org/10.1088/1674-
1056/23/8/088102.
[7] T.S. Rebelo, C.M. Pereira, M.G.F. Sales, J.P. Noronha, J. Costa-Rodrigues, F. Silva,
M. Fernandes, Anal. Chim. Acta 850 (2014) 26, https://doi.org/10.1016/j.
aca.2014.08.005.
[8] S. Lowell, J.E. Shields, Powder surface area and porosity. Book Powder surface area
and porosity, Springer Science & Business Media, 2013.
[9] P. Sharma, D.K. Singh, V. Gupta, B. Asthana, P. Mishra, R.K. Singh, Spectrochim.
Acta Part A 116 (2013), https://doi.org/10.1016/j.saa.2013.06.041.
[10] V.J. Kim, C.K. Okano, C.R. Osborne, D.M. Frank, C.T. Meana, M.S. Castaneto, Drug
Test, Anal. 11 (2019), https://doi.org/10.1002/dta.2497.
[11] R.P. Buck, E. Lindner, Pure Appl, Chem. 66 (1994), https://doi.org/10.1351/
pac199466122527.
[12] V. Yamkamon, B. Phakdee, S. Yainoy, T. Suksrichawalit, T. Tatanandana,
P. Sangkum, W. Eiamphungporn, EXCLI J. 17 (2018) 467, https://doi.org/
10.17179/excli2018-1245.
[13] U. Lad, G.M. Kale, R. Bryaskova, J. Electrochem. Soc. 161 (2014), https://doi.org/
10.1149/2.018405jes.
[14] S.B. Revin, Int. J. Chem Pharm. Sci. 3 (2015). https://www.pharmaresearchlibrar
y.com/wp-content/uploads/2016/01/IJCPS2856.pdf.
[15] H. Yang, J. Wang, C. Yang, X. Zhao, S. Xie, Z. Ge, J. Electrochem. Soc. 165 (2018),
https://doi.org/10.1149/2.1231805jes.