Facile immobilization of cholesterol oxidase on Pt,Ru–C nanocomposite and ionic liquid–modified carbon paste electrode for an efficient amperometric free cholesterol biosensing
Abstract
In present work, the enzyme cholesterol oxidase (ChOx) was immobilized by Nafion® (Naf) on Pt,Ru–C nanocomposite and an ionic liquid (IL)–modified carbon paste electrode (CPE) in order to create cholesterol biosensor (Naf/ChOx/Pt,Ru–C/IL-CPE). The prepared working electrodes were characterized using scanning electron microscopy–energy-dispersive spectrometry, while their electrochemical performance was evaluated using electrochemical impedance spectroscopic, cyclic voltammetric, and amperometric techniques. Excellent synergism between IL 1-allyl-3-methylimidazolium dicyanamide ([AMIM][DCA]), Pt,Ru–C, and ChOx, as modifiers of CPE, offers the most pronounced analytical performance for improved cholesterol amperometric determination in phosphate buffer solution pH 7.50 at a working potential of 0.60 V. Under optimized experimental conditions, a linear relationship between oxidation current and cholesterol concentration was found for the range from 0.31 to 2.46 µM, with an estimated detec...tion limit of 0.13 µM and relative standard deviation (RSD) below 5.5%. The optimized amperometric method in combination with the developed Naf/ChOx/Pt,Ru–C/IL-CPE biosensor showed good repeatability and high selectivity towards cholesterol biosensing. The proposed biosensor was successfully applied to determine free cholesterol in a human blood serum sample via its enzymatic reaction product hydrogen peroxide despite the presence of possible interferences. The percentage recovery ranged from 99.08 to 102.81%, while RSD was below 2.0% for the unspiked as well as the spiked human blood serum sample. The obtained results indicated excellent accuracy and precision of the method, concluding that the developed biosensor can be a promising alternative to existing commercial cholesterol tests used in medical practice. Graphical abstract: [Figure not available: see fulltext.]. © 2023, The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.
Keywords:
Cholesterol / Cholesterol oxidase / Electrochemical biosensor / Human blood serum sample / Ionic liquids / Pt,Ru–C nanocompositeSource:
Analytical and Bioanalytical Chemistry, 2023Funding / projects:
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200125 (University of Novi Sad, Faculty of Science) (RS-MESTD-inst-2020-200125)
- CEEPUSIII [CZ0212–16-2223]
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Institution/Community
VinčaTY - JOUR AU - Mutić, Sanja AU - Stanković, Dalibor AU - Konya, Zoltan AU - Anojčić, Jasmina PY - 2023 UR - https://vinar.vin.bg.ac.rs/handle/123456789/11370 AB - In present work, the enzyme cholesterol oxidase (ChOx) was immobilized by Nafion® (Naf) on Pt,Ru–C nanocomposite and an ionic liquid (IL)–modified carbon paste electrode (CPE) in order to create cholesterol biosensor (Naf/ChOx/Pt,Ru–C/IL-CPE). The prepared working electrodes were characterized using scanning electron microscopy–energy-dispersive spectrometry, while their electrochemical performance was evaluated using electrochemical impedance spectroscopic, cyclic voltammetric, and amperometric techniques. Excellent synergism between IL 1-allyl-3-methylimidazolium dicyanamide ([AMIM][DCA]), Pt,Ru–C, and ChOx, as modifiers of CPE, offers the most pronounced analytical performance for improved cholesterol amperometric determination in phosphate buffer solution pH 7.50 at a working potential of 0.60 V. Under optimized experimental conditions, a linear relationship between oxidation current and cholesterol concentration was found for the range from 0.31 to 2.46 µM, with an estimated detection limit of 0.13 µM and relative standard deviation (RSD) below 5.5%. The optimized amperometric method in combination with the developed Naf/ChOx/Pt,Ru–C/IL-CPE biosensor showed good repeatability and high selectivity towards cholesterol biosensing. The proposed biosensor was successfully applied to determine free cholesterol in a human blood serum sample via its enzymatic reaction product hydrogen peroxide despite the presence of possible interferences. The percentage recovery ranged from 99.08 to 102.81%, while RSD was below 2.0% for the unspiked as well as the spiked human blood serum sample. The obtained results indicated excellent accuracy and precision of the method, concluding that the developed biosensor can be a promising alternative to existing commercial cholesterol tests used in medical practice. Graphical abstract: [Figure not available: see fulltext.]. © 2023, The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature. T2 - Analytical and Bioanalytical Chemistry T1 - Facile immobilization of cholesterol oxidase on Pt,Ru–C nanocomposite and ionic liquid–modified carbon paste electrode for an efficient amperometric free cholesterol biosensing DO - 10.1007/s00216-023-04847-9 ER -
@article{ author = "Mutić, Sanja and Stanković, Dalibor and Konya, Zoltan and Anojčić, Jasmina", year = "2023", abstract = "In present work, the enzyme cholesterol oxidase (ChOx) was immobilized by Nafion® (Naf) on Pt,Ru–C nanocomposite and an ionic liquid (IL)–modified carbon paste electrode (CPE) in order to create cholesterol biosensor (Naf/ChOx/Pt,Ru–C/IL-CPE). The prepared working electrodes were characterized using scanning electron microscopy–energy-dispersive spectrometry, while their electrochemical performance was evaluated using electrochemical impedance spectroscopic, cyclic voltammetric, and amperometric techniques. Excellent synergism between IL 1-allyl-3-methylimidazolium dicyanamide ([AMIM][DCA]), Pt,Ru–C, and ChOx, as modifiers of CPE, offers the most pronounced analytical performance for improved cholesterol amperometric determination in phosphate buffer solution pH 7.50 at a working potential of 0.60 V. Under optimized experimental conditions, a linear relationship between oxidation current and cholesterol concentration was found for the range from 0.31 to 2.46 µM, with an estimated detection limit of 0.13 µM and relative standard deviation (RSD) below 5.5%. The optimized amperometric method in combination with the developed Naf/ChOx/Pt,Ru–C/IL-CPE biosensor showed good repeatability and high selectivity towards cholesterol biosensing. The proposed biosensor was successfully applied to determine free cholesterol in a human blood serum sample via its enzymatic reaction product hydrogen peroxide despite the presence of possible interferences. The percentage recovery ranged from 99.08 to 102.81%, while RSD was below 2.0% for the unspiked as well as the spiked human blood serum sample. The obtained results indicated excellent accuracy and precision of the method, concluding that the developed biosensor can be a promising alternative to existing commercial cholesterol tests used in medical practice. Graphical abstract: [Figure not available: see fulltext.]. © 2023, The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.", journal = "Analytical and Bioanalytical Chemistry", title = "Facile immobilization of cholesterol oxidase on Pt,Ru–C nanocomposite and ionic liquid–modified carbon paste electrode for an efficient amperometric free cholesterol biosensing", doi = "10.1007/s00216-023-04847-9" }
Mutić, S., Stanković, D., Konya, Z.,& Anojčić, J.. (2023). Facile immobilization of cholesterol oxidase on Pt,Ru–C nanocomposite and ionic liquid–modified carbon paste electrode for an efficient amperometric free cholesterol biosensing. in Analytical and Bioanalytical Chemistry. https://doi.org/10.1007/s00216-023-04847-9
Mutić S, Stanković D, Konya Z, Anojčić J. Facile immobilization of cholesterol oxidase on Pt,Ru–C nanocomposite and ionic liquid–modified carbon paste electrode for an efficient amperometric free cholesterol biosensing. in Analytical and Bioanalytical Chemistry. 2023;. doi:10.1007/s00216-023-04847-9 .
Mutić, Sanja, Stanković, Dalibor, Konya, Zoltan, Anojčić, Jasmina, "Facile immobilization of cholesterol oxidase on Pt,Ru–C nanocomposite and ionic liquid–modified carbon paste electrode for an efficient amperometric free cholesterol biosensing" in Analytical and Bioanalytical Chemistry (2023), https://doi.org/10.1007/s00216-023-04847-9 . .