Reagentless biosensor for phenolic compounds based on tyrosinase entrapped within gelatine film.
A simple and new reagentless phenolic compound biosensor was constructed with tyrosinase immobilized in the gelatine matrix cross-linked with formaldehyde. The morphologies of gelatine and gelatine/tryosinase were characterized by SEM. The tyrosinase retains its bioactivity when being immobilized by the gelatine film. Phenolic compounds were determined by the direct reduction of biocatalytically liberated quinone at -0.1 V vs SCE. The process parameters for the fabrication of the enzyme electrode were studied. Optimization of the experimental parameters has been performed with regard to pH, operating potential, temperature and storage stability. This biosensor exhibits a fast amperometric response to phenolic compounds. The linear range for catechol, phenol, and p-Cresol determination was from 5 x 10(-8) to 1.4 x 10(-4) M, 5 x 10(-8) to 7.1 x 10(-5)M, and 1 x 10(-7) to 3.6 x 10(-5)M, with a detection limit of 2.1 x 10(-8) M, 1.5 x 10(-8) M, and 7.1 x 10(-8 )M, respectively. The enzyme electrode retained ca.77% of its activity after 7 days of storage at 4 degrees C in a dry state. The proposed sensor presented good repeatability, evaluated in terms of relative standard deviation (R.S.D.=8.6%) for eight different biosensors and was applied for determination in water sample. The recovery for the sample was from 99.0% to 99.8%.
Li
Nan
N
Department of Chemistry, Lab of Life Analytical Chemistry, Nanjing University, Nanjing, 210093, PR China. jjzhu@nju.edu.cn
Xue
Min-Hua
MH
Yao
Hui
H
Zhu
Jun-Jie
JJ
D016227:Benzoquinones / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods*; D002396:Catechols / Q000737:chemistry; D004566:Electrodes; D004800:Enzymes, Immobilized; D005780:Gelatin / Q000737:chemistry* / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D018625:Microscopy, Atomic Force / Q000379:methods; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D010636:Phenols / Q000032:analysis*; D011199:Potentiometry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors
D016227:Benzoquinones; D002396:Catechols; D004800:Enzymes, Immobilized; D010636:Phenols; C015233:gelatin film; C004532:quinone; D005780:Gelatin; D014442:Monophenol Monooxygenase; C034221:catechol