LAKTAT TAYİNİ İÇİN LAKTAT OKSİDAZ TEMELLİ AMPEROMETRİK ENZİM ELEKTROT HAZIRLANMASI
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Dosyalar
Tarih
2014
Yazarlar
Dergi Başlığı
Dergi ISSN
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Yayıncı
Namık Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/embargoedAccess
Özet
Laktik asidin hızlı ve güvenilir analizi, klinik analizler, gıda endüstrisi ve spor hekimliği gibi alanlarda önemli bir rol oynar. Laktik asit tayini için HPLC, iyon kromatografisi veya enzimatik kitler kullanılmaktadır ancak bu yöntemler genellikle pahalı, zaman alıcı ve zahmetlidir. Laktik asidin doğrudan hızlı, ucuz, seçici ve güvenilir tayini için biyosensör kullanımı diğer tayin yöntemlerinin iyi bir alternatifidir.. Bu çalışmada laktik asit tayini için benzo[c]sinnolin, laktat oksidaz ve çok duvarlı karbon nanotüp (ÇDKNT) ile modifiye edilmiş karbon pasta enzim elektrot geliştirildi. Elektrodun çözelti/elektrot arayüzeyindeki elektron aktarım özellikleri dönüşümlü voltametri ve elektrokimyasal empedans spektroskopisi yöntemleri kullanılarak incelendi. Daha önce yapılan elektrokimyasal çalışmalar sonucunda medyatör gibi etki gösterebileceği düşünülen benzo[c]sinnolin, bu çalışmada laktik asit tayini için yeni bir medyatör olarak araştırıldı ve laktat oksidaz için iyi bir elektron alıcısı (medyatör) olduğu ve laktik asit tayininin -0,40 V (Ag/AgCl‘a karşı) gibi düşük bir potansiyelde gerçekleşmesini sağladığı görüldü. Hazırlanan enzim elektrodun optimum elektrot bileşimi ve en iyi çalışma koşulları araştırıldı. %16 benzo[c]sinnolin, %24 ÇDKNT ve %60 grafit tozu ile hazırlanan enzim elektrodun pH‘sı 7,0 olan 0,05 M fosfat tamponunda laktik aside en iyi cevabı verdiği bulundu. Bu şartlar altında enzim elektrodun doğrusal çalışma aralığı, tekrar üretilebilirlik, tekrarlanabilirlik, cevap zamanı, raf ömrü gibi performans faktörleri incelendi. Çalışmada ayrıca enzim elektrodun serumda laktik asit tayinine uygulanabilirliği araştırıldı. Geliştirilen enzim elektrot ile geniş bir derişim aralığında, girişim etkisi olmadan, tekrarlanabilirliği yüksek ve hızlı laktik asit tayini gerçekleştirildi.
Fast and reliable analysis of lactic acid plays an important role in fields such as sports medicine, clinical analysis and food industry. HPLC, ion chromatography or enzymatic kits are used for lactic acid analysis but these methods are often expensive, time consuming and laborious. The use of biosensors for direct, rapid, cheap, reliable and selective lactic acid determination is a good alternative to other detection methods. In this study for the determination of lactic acid, benzo [c] cinnoline, lactate oxidase and multi-walled carbon nanotube (MWCNT) modified carbon paste electrode was developed for the determination of lactic acid. Electron transfer properties of the electrode at solution/electrode interface were investigated using impedance spectroscopy and cyclic voltammetry methods. According to the previous electrochemical studies benzo [c] cinnoline, was investigated as a new mediator for lactic acid determination and it was used as an useful electron acceptor (mediator) for lactate oxidase and allowed lactic acid determination to be carried out at -0.40 V (Ag / AgCl against). The optimum electrode composition and the best working conditions of the constucted enzyme electrode were investigated. Enzyme electrode prepared by 16% benzo [c] cinnoline, 24% MWCNT and 60% graphite powder was found to give the best response to lactic acid in 0.05 M phosphate buffer at pH 7.0. Performance factors of the enzyme electrode such as linear working range reproducibility, repeatability, response time, shelf-life were determined under optimal working conditions. The applicability of the enzyme electrode was also evaluated in this study. Rapid and high reproducible lactic acid determination was performed with the constructed enzyme electrode in a wide concertration range with no interference effects.
Fast and reliable analysis of lactic acid plays an important role in fields such as sports medicine, clinical analysis and food industry. HPLC, ion chromatography or enzymatic kits are used for lactic acid analysis but these methods are often expensive, time consuming and laborious. The use of biosensors for direct, rapid, cheap, reliable and selective lactic acid determination is a good alternative to other detection methods. In this study for the determination of lactic acid, benzo [c] cinnoline, lactate oxidase and multi-walled carbon nanotube (MWCNT) modified carbon paste electrode was developed for the determination of lactic acid. Electron transfer properties of the electrode at solution/electrode interface were investigated using impedance spectroscopy and cyclic voltammetry methods. According to the previous electrochemical studies benzo [c] cinnoline, was investigated as a new mediator for lactic acid determination and it was used as an useful electron acceptor (mediator) for lactate oxidase and allowed lactic acid determination to be carried out at -0.40 V (Ag / AgCl against). The optimum electrode composition and the best working conditions of the constucted enzyme electrode were investigated. Enzyme electrode prepared by 16% benzo [c] cinnoline, 24% MWCNT and 60% graphite powder was found to give the best response to lactic acid in 0.05 M phosphate buffer at pH 7.0. Performance factors of the enzyme electrode such as linear working range reproducibility, repeatability, response time, shelf-life were determined under optimal working conditions. The applicability of the enzyme electrode was also evaluated in this study. Rapid and high reproducible lactic acid determination was performed with the constructed enzyme electrode in a wide concertration range with no interference effects.
Açıklama
Anahtar Kelimeler
Laktik asit, amperometri, benzo[c]sinnolin, çok duvarlı karbon nanotüp, karbon pasta, Lactic acid, amperometry, benzo [c] cinnolin, multi walled carbon nanotubes, carbon paste
