Voltammetric Determination of Ethanol by Using Mushroom Agaricus bisporus Tissue Homogenate-Based Biosensor ​

Abstract

Anew biosensor based on mushroom Agaricus bisporus tissue homogenate was developed for voltammetric determination of ethanol. Mushroom Agaricus bisporus was homogenated and immobilized onto the glassy carbon electrode via gelatine and cross-linked by glutaraldehyde to construct an electrochemical tissue homogenate based biosensor system. The homogenate contains alcohol oxidase E.C 1.1.3.13 enzyme and it catalysis the degredation of ethanol to acetaldehyde and hydrogen peroxide. Assay principle of the biosensor based on the detection of reduction peaks of oxygen by using cyclic voltammetry at potentials between 0.0 and +0.7 V. To obtain a standard graphe for ethanol differential pulse DP method was used at potentials between +0.1 and +0.4 V. The electrode response depends linearly on ethahol concentration between 1.0 and 10.0 mM. Phosphate buffer pH 7.0, 50 mM and 30°C were obtained as the optimum conditions. In characterization studies of the biosensor some parameters such as; repeatability, substrate specificity, interference effects of some substances on the biosensor response were investigated. Finally, the proposed system was applied to ethanol detection in different wine and beer samples.

Keywords

• Voltammetry,biosensor,tissue-homogenate,ethanol

Mantar Agaricus bisporus Doku Homojenati Temelli Biyosensör Kullanarak Etanolün Voltametrik Tayini

Abstract

Bu çalışmada etanolün tayini için mantar Agaricus bisporus doku homojenatı temelli yeni bir biosensor geliştirildi. Mantar Agaricus bisporus homojenize edildi ve elektrokimyasal doku homojenatı temelli bir biyosensör sistemi geliştirmek için camsı karbon elektrot üzerinde jelatin ile immobilize edilip glutaraldehit ile çapraz bağlandı. Homojenat alkol oksidaz enzimi EC 1.1.3.13 içerir ve bu enzim etanolün hidrojen peroksit ve asetaldehite yıkımını katalizler. Biosensörün ölçüm prensibi 0.0 ve +0.7 V potansiyel aralığında döngüsel voltammetri kullanılarak oksijenin indirgenme pikinin belirlenmesine dayanmaktadır. Etanol için standart grafik eldesinde +0.1 ve +0.4 V potansiyel aralığında diferansiyel puls DP metodu kullanıldı. Elektrodun cevabı 1.0 ile 10.0 mM etanol konsantrasyonu aralığında doğrusallık gösterdi. Optimum koşullar olarak pH 7.0 50 mM fosfat tamponu ve 30oC elde edildi. Biyosensörün karakterizasyonu çalışmalarında tekrarlanabilirlik, substrat spesifikliği ve bazı maddelerin biyosensör cevabı üzerine girişim etkileri gibi birtakım parametreler incelendi. Son olarak, geliştirilen sistem, farklı bira ve şarap örneklerinde etanol tayinine uygulandı

Keywords

• Voltametri,biyosensör,doku homojenatı,etanol

References

1. M. Teke, M.K. Sezgintürk, E. Dinçkaya, A. Telefoncu, Two Biosensors for Phenolic Compounds Based on Mushroom (Agaricus bisporus) Homogenate: Comparison in Terms of Some Important Parameters of the Biosensors, Prep. Biochem. Biotechnol., 38 (2008) 51.
2. J. Razumiene, V. Gureviciene, V. Laurinavicius, J.V. Grazulevicius, Amperometric detection of glucose and ethanol in beverages using flow cell and immobilised on screen-printed carbon electrode PQQ-dependent glucose or alcohol dehydrogenases, Sens. Act., B 78 (2001) 243.
3. E. Akyilmaz, E. Dinckaya, A mushroom (Agaricus bisporus) tissue homogenate based alcohol oxidase electrode for alcohol determination in serum, Talanta, 53 (2000) 505.
4. A.W. Shi, F.L. Qu, M.H. Yang, G.L. Shen, R.Q. Yu, Amperometric H2O2 biosensor based on poly- thionine nanowire/HRP/nano-Au-modified glassy carbon electrode, Sens. Act.B, 129 (2008) 779.
5. G. Wen, Y. Zhang, S. Shuang, C. Dong, M.M.F. Choi, Application of a biosensor for monitoring of ethanol, Biosens. Bioelectron., 23 (2007) 121.
6. E. Dinçkaya, E. Akyilmaz, A. Telefoncu, S. Akgöl, Eggplant tissue homogenate-based bioselective membrane electrode for determination of catechol, Ind. J. Biochem. Biophys., 36 (1999) 36.
7. D.C. Wijesuriya, and G.A. Rechnitz, Biosensors based on plant and animal tissues, Biosens. Bioelectron., 8 (1993) 155.
8. J.S. Sidwell, G.A. Rechnitz, Progress and challenges for biosensors using plant tissue materials, Biosensors, 2 (1986) 221.

9. F. Schubert, R. Renneberg, F.Scheller, L. Kirstein, Plant tissue hybrid electrode for determination of phosphate and fluoride, Anal. Chem., 56 (1984) 1677.
10. N. Smit, G.A. Rechnitz, Leaf based biocatalytic membrane electrodes, Biotechnol. Lett., 6 (1984) 209.
11. F. Schubert, U. Wollenberger, F. Scheller, Plant tissue- based amperometric tyrosine electrode, Biotechnol. Lett., 5 (1983) 239.
12. N.G. Patel, S. Meier, K. Cammann, G.C. Chemnitius, Screen-printed biosensors using different alcohol oxidases, Sens. Act. B., 75 (2001) 101.
13. D. Çökeliler, M. Mutlu, Performance of amperometric alcohol electrodes prepared by plasma polymerization technique, Anal. Chim. Acta, 469 (2002) 217.
14. M. Revenga-Parra , T. Garcia , E. Lorenzo , F. Pariente, Electrocatalytic oxidation of methanol and other short chain aliphatic alcohols on glassy carbon electrodes modified with conductive films derived from NiII-(N,N’-bis(2,5-dihydroxybenzylidene)-1,2-di- aminobenzene, Sens. Act. B.,130 (2008) 730.
15. E. Akyilmaz , E. Dinckaya , Development of a catalase based biosensor for alcohol determination in beer samples, Talanta, 61 (2003) 113.
16. M. Yonamine, N. Tawil, R. Lucia de Moraes Moreau, O. Alves Silva, Solid-phase micro-extraction-gas chromatography-mass spectrometry and headspace- gas chromatography of tetrahydrocannabinol, amphetamine, methamphetamine, cocaine and ethanol in saliva samples, J. Chromatogr. B., 789 (2003) 73.
17. T. Yarita, R. Nakajima, S. Otsuka, T. Ihara, A. Takatsu, M. Shibukawa, Determination of ethanol in alcoholic beverages by high-performance liquid chromatography-flame ionization detection using pure water as mobile phase, J. Chromatogr. A, 976 (2002) 387.
18. M.J.L. Castanon, A.J. Miranda Ordieres, P. Tunon Blanco, Amperometric detection of ethanol with poly- (o-phenylenediamine) modified enzyme electrodes, Biosens. Bioelectron., 12 (1997) 511.
19. M. Boujtita , J.P. Hart, R. Pittson, Development of a disposable ethanol biosensor based on a chemically modified screen-printed electrode coated with alcohol oxidase for the analysis of beer, Biosens. Bioelectron., 15 (2000) 257.
20. A.M. Azevedo, D.M.F. Prazeres, J.M.S. Cabral, L.P. Fonseca, Ethanol sensors based on alcohol oxidase, Biosens. Bioelectron., 21 (2005) 235.