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Keywords = monoenzyme biosensor

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19 pages, 3696 KiB  
Article
Adaptation of Conductometric Monoenzyme Biosensor for Rapid Quantitative Analysis of L-arginine in Dietary Supplements
by Olga Y. Saiapina, Kseniia Berketa, Andrii S. Sverstiuk, Lyubov Fayura, Andriy A. Sibirny, Sergei Dzyadevych and Oleksandr O. Soldatkin
Sensors 2024, 24(14), 4672; https://doi.org/10.3390/s24144672 - 18 Jul 2024
Cited by 3 | Viewed by 834
Abstract
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 [...] Read more.
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. Full article
(This article belongs to the Section Biosensors)
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Figure 1
<p>Portable conductometric analyzer used for the biosensor measurements and typical conductometric transducer.</p> Full article ">Figure 2
<p>Dependence of biosensor responses on pH (<b>a</b>) and buffer capacity (<b>b</b>) of the working buffer solution. A: The concentration of L-arginine is 1.5 mM. The study of the effect of pH was carried out in 2.5 mM Polymix buffer solution. B: 1—1 mM buffer, 2—2.5 mM buffer, 3—5 mM buffer, 4—10 mM buffer, 5—20 mM buffer. The analysis of the effect of buffer capacity was carried out in phosphate buffer with pH 6.2.</p> Full article ">Figure 3
<p>Dependence of the response of the biosensor on the temperature (<b>a</b>), protein concentration (<b>b</b>), and ionic strength (<b>c</b>) of the solution in the measuring cell. The concentration of L-arginine is 1.5 mM. Measurements were carried out in 5 mM phosphate buffer with pH 6.2.</p> Full article ">Figure 4
<p>An example of a real biosensor response to the addition of a small concentration of analyte and the principle of the LOD calculation.</p> Full article ">Figure 5
<p>Calibration curve with an indication of its linear section. Measurements were performed in 5 mM phosphate buffer, pH 6.2.</p> Full article ">Figure 6
<p>Operational stability of the biosensor ADI-based biosensor during 14 days of use. Biosensor responses to 1.5 mM L-arginine in 5 mM phosphate buffer solution, pH 6.2.</p> Full article ">Figure 7
<p>Calculation of L-arginine concentration in the measuring cell using the method of standard additions.</p> Full article ">Figure 8
<p>Repeatability of results of measurements in real samples via biosensor-based ADI. Measurement in 5 mM phosphate buffer, pH 6.2 (method of standard additions (500-fold dilution)).</p> Full article ">Figure 9
<p>Comparison of L-arginine determination data in dietary supplement samples obtained using the biosensor method (n = 5) and control methods. Samples of dietary supplements: 1—“L-arginine 500 mg” (“Solgar Vitamin and Herb”); 2—“L-arginine 500 mg” (“Now Foods”); 3—“L-arginine” (LLC “Elit-Pharm”).</p> Full article ">Figure 10
<p>Correlation between data of the biosensor determination of L-arginine content in samples of dietary supplements and data of ion-exchange chromatography (<b>a</b>) and data of the spectrophotometric analysis (<b>b</b>). Biosensor determination was carried out in 5 mM phosphate buffer, pH 6.2.</p> Full article ">
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