Biopolym. Cell. 2017; 33(2):124-134.
http://dx.doi.org/10.7124/bc.00094A
Molecular and Cell Biotechnologies
Highly selective amperometric biosensor for uric acid determination in real samples
1Zinchenko O. A., 1Shkotova L. V., 2Kulynych T. U., 3Zinkina O. O., 1, 3Soldatkin A. P.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. Taras Shevchenko National University of Kyiv
    64, Volodymyrska Str., Kyiv, Ukraine, 01601
  3. Institute of High Technologies,
    Taras Shevchenko National University of Kyiv
    2, korp.5, Pr. Akademika Hlushkova, Kyiv, Ukraine, 03022

Abstract

Aim. To develop an amperometric biosensor based on immobilized uricase (1.7.3.3) from Arthrobacter Globiformis and a platinum disk electrode for the detection of uric acid in biological fluids. Methods. To obtain a highly selective detection of the uric acid concentration, an additive semi-permeable polymer film was formed on the surface of a platinum disk electrode by electro-polymerisation of m-phenylene diamine. The enzymatic selective layer was formed on the poly-m-phenylene diamine membrane using uricase immobilized in BSA matrix by a non-toxic crosslinking agent – poly(ethylene glycol) diglycidyl ether (PEGDE). Results. An influence of possible interfering substances – ascorbic acid, cysteine, urea, glucose, glutamic acid and lactic acid – was studied. Almost no effect of these electrochemical compounds on the biosensor response was found, indicating that the selectivity of the developed biosensor is very high. The biosensor characteristics were determined: detection limit 0.001 mM (s/n = 3), linear working range 0.008–0.218 mM, sensitivity 165 μA·mM–1 cm–2. The biosensor stability and reproducibility were studied and shown. Conclusions. The developed biosensor was validated by comparing the results of the urine samples analysis provided with the biosensor and the spectrophotometric method (correlation coefficient r = 0.99).This biosensor is found to be promising method for uric acid detection in the real samples.
Keywords: uricase, amperometric biosensor, uric acid, m-phenylene diamine, poly(ethylene glycol) diglycidyl ether.
Full text: (PDF, in English)

References

[1] Lesch M, Nyhan WL. A Familial disorder of uric acid metabolism and central nervous system function. Am J Med. 1964;36:561–70.
[2] Maksudova A, Salikhov I, Habirov R, Halfina T. The gout. Moscow: Medpress-inform, 2012. 96 p.
[3] Heinig M, Johnson RJ. Role of uric acid in hypertension, renal disease, and metabolic syndrome. Cleve Clin J Med. 2006;73(12):1059-64.
[4] Alderman M, Aiyer KJ. Uric acid: role in cardiovascular disease and effects of losartan. Curr Med Res Opin. 2004;20(3):369-79.
[5] Raj C, Ohsaka T. Voltammetric detection of uric acid in the presence of ascorbic acid at a gold electrode modi-fied with a self-assembled monolayer of heteroaromatic thiol. J Electroanal Chem. 2003; 540: 69–77.
[6] Isdale IC, Buchanan MJ, Rose BS. Serum uric acid estimation. A modified chemical method. Ann Rheum Dis. 1966;25(2):184–5.
[7] Lorentz K, Berndt W. Enzymic determination of uric acid by a colorimetric method. Anal Biochem. 1967; 18(1):58–63.
[8] Hong H-C, Huang H-J. Flow injection analysis of uric acid with a uricase-and horseradish peroxidase-coupled Sepharose column based luminol chemiluminescence system. Anal Chim Acta. 2003, 499(1-2):41–6.
[9] He D, Zhang Z, Huang Y, Hu Y, Zhou H, Chen D. Chemiluminescence microflow injection analysis system on a chip for the determination of uric acid without enzyme. Luminescence. 2005;20(4–5):271–5.
[10] Galbán J, Andreu Y, Almenara MJ, de Marcos S, Castillo JR. Direct determination of uric acid in serum by a fluorometric-enzymatic method based on uricase. Talanta. 2001;54(5):847–54.
[11] Abdullin IF, Bakanina YuN, Turova EN, Budnikov GK. Determination of uric acid by voltammetry and coulometric titration. J Anal Chem. 2001; 56(5): 45356.
[12] Rocha DL, Rocha FRP. A flow-based procedure with solenoid micro-pumps for the spectrophotometric determina-tion of uric acid in urine. Microchem J. 2010; 94(1):53–9.
[13] Perelló J, Sanchis P, Grases F. Determination of uric acid in urine, saliva and calcium oxalate renal calculi by high-performance liquid chromatography/mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2005;824(1–2):175–80.
[14] Dai X, Fang X, Zhang C, Xu R, Xu B. Determination of serum uric acid using high-performance liquid chroma-tography (HPLC)/isotope dilution mass spectrometry (ID-MS) as a candidate reference method. J Chromatogr B Analyt Technol Biomed Life Sci. 2007;857(2):287–95.
[15] Xu DK, Hua L, Li ZM, Chen HY. Identification and quantitative determination of uric acid in human urine and plasma by capillary electrophoresis with amperometric detection. J Chromatogr B Biomed Sci Appl. 1997;694(2):461–6.
[16] Soldatkin AP, Dzyadevych SV, Korpan YI, Sergeyeva TA, Arkhypova VN, Biloivan OA, Soldatkin OA, Shkotova LV, Zinchenko OA, Peshkova VM, Saiapina OY, Marchenko SV, El’Skaya AV. Biosensors. A quarter of a century of R&D experience. Biopolym Cell. 2013; 29(3):188–206.
[17] Vasylieva N, Marinesco S, Enzyme Immobilization on Microelectrode Biosensors, in: Microelectrode Biosensors. Eds. Marinesco S, Dale N, Neuromethods, 2013: 95-114.
[18] Vasylieva N, Maucler C, Meiller A, Viscogliosi H, Lieutaud T, Barbier D, Marinesco S. Immobilization method to preserve enzyme specificity in biosensors: consequences for brain glutamate detection. Anal Chem. 2013;85(4):2507–15.
[19] Shkotova LV, Slast'ia EA, Zhyliakova TA, Soldatkin OP, Schuhmann W, Dziadevych SV. [Amperometric biosensor for ethanol analysis in wines and grape must during wine fermentation]. Ukr Biokhim Zh (1999). 2005;77(1):96-103.
[20] Schuvailo OM, Soldatkin OO, Lefebre A, Cespuglio R, Soldatkin AP. Highly selective microbiosensors for in vivo measurement of glucose, lactate and glutamate. Anal Chim Acta. 2006; 573–574:110–6.
[21] Goriushkina TB, Shkotova LV, Gayda GZ, Klepach HM, Gonchar MV, Soldatkin AP, Dzyadevych SV. Amperometric biosensor based on glycerol oxidase for glycerol determination. Sens Actuators B. 2010, 144(2):361–7.
[22] Lurie Yu. Handbook of analytical chemistry. Moscow: Chemistry, 1967.
[23] Trevan M. Immobilized enzymes: An introduction and applications in biotechnology. John Wiley & Sons, Chichester. 1980.
[24] Zhao Y, Yang X, Lu W, Liao H, Liao F. Uricase based methods for determination of uric acid in serum. Micro-chim Acta. 2009; 164(1-2):1–6.