An amperometric hand-held Nitrate biosensor based on Nitrate reductase immobilized on a thin film electrode: a tool for in-filed water and food analysis
Document Type : Original research
Authors
1 Bioprocessing and Biodetection Lab (BBL), Department of Food Science, Technology, and Engineering, University of Tehran, Karaj, Iran.
2 Bioprocessing and Biodetection Lab (BBL), Department of Food Science, Technology, and Engineering, University of Tehran, Karaj, Iran
10.22059/jfabe.2024.373543.1167
Abstract
This study presents the design and evaluation of a portable amperometric biosensor for detecting nitrates, utilizing a gold three-electrode system with immobilized nitrate reductase. With a sensitivity of 12.01 μA mM−1 cm−2 over a 0.16–0.96 mM range and a regression coefficient of 0.98, the biosensor outperforms traditional UV spectrophotometric methods (0.01–0.19 mM) by offering a broader range (0.16–0.96 mM) and greater accuracy, as evidenced by a 6.69% relative standard deviation for reproducibility and a detection limit of 0.14 mM. Maintaining 63% of its initial response after four days and showing minimal interference from nitrite (1 mM), this biosensor is highly suitable for water testing, although pre-treatment is required for analyzing food samples due to phenolic compounds.
Keywords
Main Subjects
References
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Bollella, Paolo, and Lo Gorton. 2018. 'Enzyme based amperometric
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Branch, Shirmir D, Amanda M Lines, John Lynch, Job M Bello, William R
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thin-film electrode chip for spectroelectrochemical sensing',
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Dai, Weiwei, Ivan Selesnick, John-Ross Rizzo, Janet Rucker, and Todd
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filter and the quantitative analysis of saccades', Journal of vision,
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Ferreyra, Nancy F, and Velia M Solı́s. 2004. 'An amperometric nitrate
reductase–phenosafranin electrode: kinetic aspects and analytical
applications', Bioelectrochemistry, 64: 61-70.
Ferreyra, NF, SA Dassie, and VM Solis. 2000. 'Electroreduction of methyl
viologen in the presence of nitrite. Its influence on enzymatic
electrodes', Journal of Electroanalytical Chemistry, 486: 126-32.
Glazier, Scott A, Ellen R Campbell, and Wilbur H Campbell. 1998.
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amperometric electrode and nitrate assay of fertilizers and
drinking water', Analytical chemistry, 70: 1511-15.
Jadán, Felipe, María-Concepción Aristoy, and Fidel Toldrá. 2017. 'Biosensor
Based on Immobilized Nitrate Reductase for the Quantification of
Nitrate Ions in Dry-Cured Ham', Food Analytical Methods, 10:
3481-86.
Kalimuthu, Palraj, Katrin Fischer-Schrader, Günter Schwarz, and Paul V
Bernhardt. 2015. 'A sensitive and stable amperometric nitrate
biosensor employing Arabidopsis thaliana nitrate reductase', JBIC
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Kalimuthu, Palraj, Tobias Kruse, and Paul V Bernhardt. 2021. 'A highly
sensitive and stable electrochemical nitrate biosensor',
Electrochimica Acta, 386: 138480.
Lee, GeonHui, JuKyung Lee, JeongHoon Kim, Hak Soo Choi, Jonghan Kim,
SangHoon Lee, and HeaYeon Lee. 2017. 'Single microfluidic
electrochemical sensor system for simultaneous multi-pulmonary
hypertension biomarker analyses', Scientific reports, 7: 7545.
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simultaneous determination of nitrite and nitrate', Biosensors and
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monolayer platform', Analyst, 137: 747-53.
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Vieira. 2008. 'Bean sprout peroxidase biosensor based on lcysteine
self-assembled monolayer for the determination of
dopamine', Sensors and actuators B: Chemical, 133: 364-69.
Monošík, Rastislav, Miroslav Streďanský, Gabriel Greif, and Ernest Šturdík.
2012. 'A rapid method for determination of L-lactic acid in real
samples by amperometric biosensor utilizing nanocomposite',
Food Control, 23: 238-44.
Muñoz-Martínez, Adrián, Omar González Peña, Jordi Colomer-Farrarons,
José Rodríguez-Delgado, Alfonso Ávila-Ortega, and Graciano
Dieck-Assad. 2018. 'Electrochemical Instrumentation of an
Embedded Potentiostat System (EPS) for a Programmable-
System-On-a-Chip', Sensors, 18: 4490.
Pilas, Johanna, Yasemen Yazici, Thorsten Selmer, Michael Keusgen, and
Michael Schöning. 2018. 'Application of a portable multi-analyte
biosensor for organic acid determination in silage', Sensors, 18:
1470.
Quan, De, Jun Ho Shim, Jong Dae Kim, Hyung Soo Park, Geun Sig Cha, and
Hakhyun Nam. 2005. 'Electrochemical determination of nitrate
with nitrate reductase-immobilized electrodes under ambient air',
Analytical chemistry, 77: 4467-73.
Reich, Peggy, Regina Stoltenburg, Beate Strehlitz, Dieter Frense, and Dieter
Beckmann. 2017. 'Development of an Impedimetric Aptasensor
for the Detection of Staphylococcus aureus', International journal
of molecular sciences, 18: 2484.
Sohail, Manzar, and Samuel B Adeloju. 2016. 'Nitrate biosensors and
biological methods for nitrate determination', Talanta, 153: 83-
98.
Strehlitz, B, B Gründig, K-D Vorlop, P Bartholmes, H Kotte, and U
Stottmeister. 1994. 'Artificial electron donors for nitrate and
nitrite reductases usable as mediators in amperometric
biosensors', Fresenius' journal of analytical chemistry, 349: 676-
78.
Uludag, Yildiz, Zehra Olcer, and Mahmut Samil Sagiroglu. 2014. 'Design
and characterisation of a thin-film electrode array with shared
reference/counter electrodes for electrochemical detection',
Biosensors and Bioelectronics, 57: 85-90.
Umar, Mohammad Faisal, and Abu Nasar. 2018. 'Reduced graphene
oxide/polypyrrole/nitrate reductase deposited glassy carbon
electrode (GCE/RGO/PPy/NR): biosensor for the detection of
nitrate in wastewater', Applied Water Science, 8: 1-10.
Wang, Qiu-Hua, Li-Ju Yu, Yang Liu, Lan Lin, Ri-gang Lu, Jian-ping Zhu,
Lan He, and Zhong-Lin Lu. 2017. 'Methods for the detection and
determination of nitrite and nitrate: A review', Talanta, 165: 709-
20.
Wang, Yusheng, Xiaoqi Wang, Wei Lu, Qingpan Yuan, Yourong Zheng, and
Bo Yao. 2019. 'A thin film polyethylene terephthalate (PET)
electrochemical sensor for detection of glucose in sweat', Talanta,
198: 86-92.
Zhang, Wen, Yiwei Xu, Haroon E Tahir, and Xiaobo Zou. 2017.
'Determinations of trace lead in various natural samples by a novel
active microband-electrode probe', Chemical Engineering
Journal, 309: 305-12.
Alahi, Md Eshrat E, and Subhas Chandra Mukhopadhyay. 2018. 'Detection
methods of nitrate in water: A review', Sensors and Actuators A:
Physical, 280: 210-21.
Amor-Gutiérrez, Olaya, EC Rama, María Teresa Fernández-Abedul, and
Agustín Costa-García. 2017. 'Bioelectroanalysis in a Drop:
Construction of a Glucose Biosensor', Journal of Chemical
Education, 94: 806-12.
Association, American Public Health, American Water Works Association,
Water Pollution Control Federation, and Water Environment
Federation. 1995. Standard methods for the examination of water
and wastewater (American Public Health Association.).
Bard, Allen J, and Larry R Faulkner. 2001. 'Fundamentals and applications',
Electrochemical Methods, 2: 482.
Ben-Yoav, Hadar, Sheryl E Chocron, Thomas E Winkler, Eunkyoung Kim,
Deanna L Kelly, Gregory F Payne, and Reza Ghodssi. 2015. 'An
electrochemical micro-system for clozapine antipsychotic
treatment monitoring', Electrochimica Acta, 163: 260-70.
Bollella, Paolo, and Lo Gorton. 2018. 'Enzyme based amperometric
biosensors', Current Opinion in Electrochemistry, 10: 157-73.
Branch, Shirmir D, Amanda M Lines, John Lynch, Job M Bello, William R
Heineman, and Samuel A Bryan. 2017. 'Optically transparent
thin-film electrode chip for spectroelectrochemical sensing',
Analytical chemistry, 89: 7324-32.
Can, Faruk, Seyda Korkut Ozoner, Pinar Ergenekon, and Elif Erhan. 2012.
'Amperometric nitrate biosensor based on Carbon
nanotube/Polypyrrole/Nitrate reductase biofilm electrode',
Materials Science and Engineering: C, 32: 18-23.
Chooto, Pipat. 2019. 'Cyclic Voltammetry and Its Applications.' in,
Voltammetry (IntechOpen).
Da Silva, S, D Shan, and S Cosnier. 2004. 'Improvement of biosensor
performances for nitrate determination using a new hydrophilic
poly (pyrrole-viologen) film', Sensors and Actuators B: Chemical,
103: 397-402.
Dai, Weiwei, Ivan Selesnick, John-Ross Rizzo, Janet Rucker, and Todd
Hudson. 2017. 'A nonlinear generalization of the Savitzky-Golay
filter and the quantitative analysis of saccades', Journal of vision,
17: 10-10.
Ferreyra, Nancy F, and Velia M Solı́s. 2004. 'An amperometric nitrate
reductase–phenosafranin electrode: kinetic aspects and analytical
applications', Bioelectrochemistry, 64: 61-70.
Ferreyra, NF, SA Dassie, and VM Solis. 2000. 'Electroreduction of methyl
viologen in the presence of nitrite. Its influence on enzymatic
electrodes', Journal of Electroanalytical Chemistry, 486: 126-32.
Glazier, Scott A, Ellen R Campbell, and Wilbur H Campbell. 1998.
'Construction and characterization of nitrate reductase-based
amperometric electrode and nitrate assay of fertilizers and
drinking water', Analytical chemistry, 70: 1511-15.
Jadán, Felipe, María-Concepción Aristoy, and Fidel Toldrá. 2017. 'Biosensor
Based on Immobilized Nitrate Reductase for the Quantification of
Nitrate Ions in Dry-Cured Ham', Food Analytical Methods, 10:
3481-86.
Kalimuthu, Palraj, Katrin Fischer-Schrader, Günter Schwarz, and Paul V
Bernhardt. 2015. 'A sensitive and stable amperometric nitrate
biosensor employing Arabidopsis thaliana nitrate reductase', JBIC
Journal of Biological Inorganic Chemistry, 20: 385-93.
Kalimuthu, Palraj, Tobias Kruse, and Paul V Bernhardt. 2021. 'A highly
sensitive and stable electrochemical nitrate biosensor',
Electrochimica Acta, 386: 138480.
Lee, GeonHui, JuKyung Lee, JeongHoon Kim, Hak Soo Choi, Jonghan Kim,
SangHoon Lee, and HeaYeon Lee. 2017. 'Single microfluidic
electrochemical sensor system for simultaneous multi-pulmonary
hypertension biomarker analyses', Scientific reports, 7: 7545.
Lopin, Prattana, and Kyle V Lopin. 2018. 'PSoC-Stat: A single chip open
source potentiostat based on a Programmable System on a Chip',
PloS one, 13: e0201353.
Madasamy, Thangamuthu, Manickam Pandiaraj, Murugesan Balamurugan,
Kalpana Bhargava, Niroj Kumar Sethy, and Chandran
Karunakaran. 2014. 'Copper, zinc superoxide dismutase and
nitrate reductase coimmobilized bienzymatic biosensor for the
simultaneous determination of nitrite and nitrate', Biosensors and
Bioelectronics, 52: 209-15.
Matharu, Zimple, Pratima R Solanki, Vinay Gupta, and BD Malhotra. 2012.
'Mediator free cholesterol biosensor based on self-assembled
monolayer platform', Analyst, 137: 747-53.
Moccelini, Sally Katiuce, Suellen Cadorin Fernandes, and Iolanda Cruz
Vieira. 2008. 'Bean sprout peroxidase biosensor based on lcysteine
self-assembled monolayer for the determination of
dopamine', Sensors and actuators B: Chemical, 133: 364-69.
Monošík, Rastislav, Miroslav Streďanský, Gabriel Greif, and Ernest Šturdík.
2012. 'A rapid method for determination of L-lactic acid in real
samples by amperometric biosensor utilizing nanocomposite',
Food Control, 23: 238-44.
Muñoz-Martínez, Adrián, Omar González Peña, Jordi Colomer-Farrarons,
José Rodríguez-Delgado, Alfonso Ávila-Ortega, and Graciano
Dieck-Assad. 2018. 'Electrochemical Instrumentation of an
Embedded Potentiostat System (EPS) for a Programmable-
System-On-a-Chip', Sensors, 18: 4490.
Pilas, Johanna, Yasemen Yazici, Thorsten Selmer, Michael Keusgen, and
Michael Schöning. 2018. 'Application of a portable multi-analyte
biosensor for organic acid determination in silage', Sensors, 18:
1470.
Quan, De, Jun Ho Shim, Jong Dae Kim, Hyung Soo Park, Geun Sig Cha, and
Hakhyun Nam. 2005. 'Electrochemical determination of nitrate
with nitrate reductase-immobilized electrodes under ambient air',
Analytical chemistry, 77: 4467-73.
Reich, Peggy, Regina Stoltenburg, Beate Strehlitz, Dieter Frense, and Dieter
Beckmann. 2017. 'Development of an Impedimetric Aptasensor
for the Detection of Staphylococcus aureus', International journal
of molecular sciences, 18: 2484.
Sohail, Manzar, and Samuel B Adeloju. 2016. 'Nitrate biosensors and
biological methods for nitrate determination', Talanta, 153: 83-
98.
Strehlitz, B, B Gründig, K-D Vorlop, P Bartholmes, H Kotte, and U
Stottmeister. 1994. 'Artificial electron donors for nitrate and
nitrite reductases usable as mediators in amperometric
biosensors', Fresenius' journal of analytical chemistry, 349: 676-
78.
Uludag, Yildiz, Zehra Olcer, and Mahmut Samil Sagiroglu. 2014. 'Design
and characterisation of a thin-film electrode array with shared
reference/counter electrodes for electrochemical detection',
Biosensors and Bioelectronics, 57: 85-90.
Umar, Mohammad Faisal, and Abu Nasar. 2018. 'Reduced graphene
oxide/polypyrrole/nitrate reductase deposited glassy carbon
electrode (GCE/RGO/PPy/NR): biosensor for the detection of
nitrate in wastewater', Applied Water Science, 8: 1-10.
Wang, Qiu-Hua, Li-Ju Yu, Yang Liu, Lan Lin, Ri-gang Lu, Jian-ping Zhu,
Lan He, and Zhong-Lin Lu. 2017. 'Methods for the detection and
determination of nitrite and nitrate: A review', Talanta, 165: 709-
20.
Wang, Yusheng, Xiaoqi Wang, Wei Lu, Qingpan Yuan, Yourong Zheng, and
Bo Yao. 2019. 'A thin film polyethylene terephthalate (PET)
electrochemical sensor for detection of glucose in sweat', Talanta,
198: 86-92.
Zhang, Wen, Yiwei Xu, Haroon E Tahir, and Xiaobo Zou. 2017.
'Determinations of trace lead in various natural samples by a novel
active microband-electrode probe', Chemical Engineering
Journal, 309: 305-12.
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