A new Biosensor Development Pencil on Based Graphite Electrode-Ascorbate Oxidase for Dtermination of L-ascorbic acid (C Vitamin)

Öz

In this study, in respect to provide an novel and inventive viewpoint, a new assay for analysis of L-ascorbic acid was enhanced using PGE. Ascorbat oxidase was crosswise bounded using gluteraldehyde and gelatine, stable to the surface of pencil graphite electrode and devised biosensor was used for the detection of ascorbic acid. L-ascorbic acid measuring were done amperometrically at -0.7 V using the improved biosensor. For the PGE/gelatine- glutaraldehyde /ascorbate oxidase biosensor ascorbate oxidase concentration, holding time in glutaradehyde, gelatine amount and glutaraldehyde stratifying repat cycle were determined to be 1.5 U/mL, 3minutes, 20 mg and 3 times stratification in turn after optimization works. Potassium phosphate buffer (pH:7, 50 mM) and 30ºC warmth determined to procure optimum labouring conditions. After the characterization labours, 25 µM - 500 µM detection space was provide to be linear for PGE/gelatine- glutaraldehyde/ascorbate oxidase biosensor. Regarding the results, the % coefficient of variation (V.K) = 0.44 and the standard deviation (S.S) = ±1.46 µM. As a result of the experiments on storage stability, it was determined that 75% activity was preserved at the end of the 4-week period.

Anahtar Kelimeler

• Pencil graphite electrode
• ascorbate oxidase
• L-ascorbic acid
• biosensör
• amperometric

L-Askorbik asit (C vitamini) Tayinine Yönelik Kalem Grafit Elektrot-Askorbat Oksidaz Temelli Yeni Bir Biyosensör Geliştirilmesi

Öz

Bu çalışmada, biyosensör teknolojisi için özgün ve yeni bir bakış açısı katmak maksadıyla PGE kullanılarak L-askorbik asit analizi için yeni bir sensör geliştirilmiştir. Askorbat oksidaz enzimi glutaraldehid ve jelatin kullanılarak çapraz bağlanmış, kalem grafit elektrot yüzeyinde tutturulmuştur ve geliştirilen biyosensör L-askorbik asit tayini için kullanılmıştır. Ölçümler amperometrik yöntem kullanılarak tüketilen oksijen miktarı ile orantısal akım değerlerindeki azalmanın belirlenmesi ile yapılmıştır. Tasarlanan biyosensör ile L-askorbik asit ölçümleri -0.7 V’ta amperometrik yöntem ile gerçekleştirilmiştir. Optimizasyon çalışmalarından PGE/jelatin- glutaraldehit/askorbat oksidaz modifiye biyosensör için askorbat oksidaz konsantrasyonu, glutaraldehitte bekletme süresi, jelatin miktarı, ve glutaraldehit tabakalandırma sayısı sırasıyla 1,5 U/mL, 3 dakika, 20 mg ve 3 kez olarak analiz edilmiştir. Kullanılan Potasyum fosfat tamponu (pH:7, 50 mM) ve 30°C’de optimum çalışma koşullarını sağladığı belirlenmiştir. PGE/jelatin- glutaraldehit/askorbat oksidaz biyosensörü için karakterizasyon çalışmalarında doğrusal tayin aralığı 25µM - 500µM bulunmuştur. Sonuçlarına ilişkin olarak % varyasyon katsayısı (V.K) = 0,44 ve standart sapma (S.S) = ±1,46 µM olarak belirlenmiştir. Depolama kararlılığına ilişkin yapılan denemeler sonucunda 4 haftalık sürecin sonunda %75’lik aktivitenin korunduğu tespit edilmiştir.

Anahtar Kelimeler

• Kalem grafit elektrot
• askorbat oksidaz
• L-askorbik asit
• biyosensör
• amperometrik

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