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Adaptation of Conductometric Monoenzyme Biosensor for Rapid Quantitative Analysis of L-arginine in Dietary Supplements
The present study reports on the development, adaptation, and optimization of a novel monoenzyme conductometric biosensor based on a recombinant arginine deiminase (ADI) for the determination of arginine in dietary supplements with a high accuracy of results. Aiming for the highly sensitive determin...
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Published in: | Sensors (Basel, Switzerland) Switzerland), 2024-07, Vol.24 (14), p.4672 |
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description | The present study reports on the development, adaptation, and optimization of a novel monoenzyme conductometric biosensor based on a recombinant arginine deiminase (ADI) for the determination of arginine in dietary supplements with a high accuracy of results. Aiming for the highly sensitive determination of arginine in real samples, we studied the effect of parameters of the working buffer solution (its pH, buffer capacity, ionic strength, temperature, and protein concentration) on the sensitivity of the biosensor to arginine. Thus, it was determined that the optimal buffer is a 5 mM phosphate buffer solution with pH 6.2, and the optimal temperature is 39.5 °C. The linear functioning range is 2.5-750 µM of L-arginine with a minimal limit of detection of 2 µM. The concentration of arginine in food additive samples was determined using the developed ADI-based biosensor. Based on the obtained results, the most effective method of biosensor analysis using the method of standard additions was chosen. It was also checked how the reproducibility of the biosensor changes during the analysis of pharmaceutical samples. The results of the determination of arginine in real samples using a conductometric biosensor based on ADI clearly correlated with the data obtained using the method of ion-exchange chromatography and enzymatic spectrophotometric analysis. We concluded that the developed biosensor would be effective for the accurate and selective determination of arginine in dietary supplements intended for the prevention and/or elimination of arginine deficiency. |
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Aiming for the highly sensitive determination of arginine in real samples, we studied the effect of parameters of the working buffer solution (its pH, buffer capacity, ionic strength, temperature, and protein concentration) on the sensitivity of the biosensor to arginine. Thus, it was determined that the optimal buffer is a 5 mM phosphate buffer solution with pH 6.2, and the optimal temperature is 39.5 °C. The linear functioning range is 2.5-750 µM of L-arginine with a minimal limit of detection of 2 µM. The concentration of arginine in food additive samples was determined using the developed ADI-based biosensor. Based on the obtained results, the most effective method of biosensor analysis using the method of standard additions was chosen. It was also checked how the reproducibility of the biosensor changes during the analysis of pharmaceutical samples. The results of the determination of arginine in real samples using a conductometric biosensor based on ADI clearly correlated with the data obtained using the method of ion-exchange chromatography and enzymatic spectrophotometric analysis. We concluded that the developed biosensor would be effective for the accurate and selective determination of arginine in dietary supplements intended for the prevention and/or elimination of arginine deficiency.</description><identifier>ISSN: 1424-8220</identifier><identifier>EISSN: 1424-8220</identifier><identifier>DOI: 10.3390/s24144672</identifier><identifier>PMID: 39066069</identifier><language>eng</language><publisher>Switzerland: MDPI AG</publisher><subject>arginine ; Arginine - analysis ; Arginine - chemistry ; arginine deiminase ; Biosensing Techniques - methods ; Biosensors ; Chromatography ; conductometric transducer ; Dietary supplements ; Dietary Supplements - analysis ; Electrodes ; Enzymes ; Hydrogen-Ion Concentration ; Hydrolases - chemistry ; Limit of Detection ; monoenzyme biosensor ; Osmolar Concentration ; real sample analysis ; Reproducibility of Results ; Temperature</subject><ispartof>Sensors (Basel, Switzerland), 2024-07, Vol.24 (14), p.4672</ispartof><rights>2024 by the authors. 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The results of the determination of arginine in real samples using a conductometric biosensor based on ADI clearly correlated with the data obtained using the method of ion-exchange chromatography and enzymatic spectrophotometric analysis. 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The results of the determination of arginine in real samples using a conductometric biosensor based on ADI clearly correlated with the data obtained using the method of ion-exchange chromatography and enzymatic spectrophotometric analysis. We concluded that the developed biosensor would be effective for the accurate and selective determination of arginine in dietary supplements intended for the prevention and/or elimination of arginine deficiency.</abstract><cop>Switzerland</cop><pub>MDPI AG</pub><pmid>39066069</pmid><doi>10.3390/s24144672</doi><orcidid>https://orcid.org/0000-0003-2915-716X</orcidid><orcidid>https://orcid.org/0000-0001-8644-0776</orcidid><oa>free_for_read</oa></addata></record> |
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subjects | arginine Arginine - analysis Arginine - chemistry arginine deiminase Biosensing Techniques - methods Biosensors Chromatography conductometric transducer Dietary supplements Dietary Supplements - analysis Electrodes Enzymes Hydrogen-Ion Concentration Hydrolases - chemistry Limit of Detection monoenzyme biosensor Osmolar Concentration real sample analysis Reproducibility of Results Temperature |
title | Adaptation of Conductometric Monoenzyme Biosensor for Rapid Quantitative Analysis of L-arginine in Dietary Supplements |
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