Amperometric biosensor based on horseradish peroxidase for biogenic amine determinations in biological samples.
An amperometric biosensor for total biogenic amine determinations, using a carbon paste electrode modified with horseradish peroxidase (HRP) enzyme is described. The HRP immobilization on graphite was made using bovine serum albumin, carbodiimide and glutaraldehyde. The biosensor response was optimized using serotonin and it presented the best performance in 0.1 mol l(-1) phosphate buffer (pH=7.0) containing 10 micromol l(-1) of hydrogen peroxide. Under optimized operational conditions at -50 mV versus SCE, a linear response range from 40 to 470 ng ml(-1) was obtained. The detection limit was 17 ng ml(-1) and the response time was 0.5s. The proposed sensor presented a stable response during 4h under continuous monitoring. The difference of the response between six sensor preparations was <2%. The sensor was applied in the determination of total biogenic amines (neurotransmitters) in rat blood samples with success, obtaining a recovery average of 102%.
Castilho
Tânia Jacometo
TJ
Instituto de Química, UNICAMP, P.O. Box 6154, 13083-970 Campinas, São Paulo, Brazil.
Sotomayor
Maria del Pilar Taboada
Mdel P
Kubota
Lauro Tatsuo
LT
D000818:Animals; D015306:Biogenic Monoamines / Q000032:analysis* / Q000097:blood; D015374:Biosensing Techniques; D002021:Buffers; D004563:Electrochemistry; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005976:Glutaral; D006735:Horseradish Peroxidase / Q000737:chemistry*; D006861:Hydrogen Peroxide; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D051381:Rats; D012701:Serotonin / Q000097:blood; D012710:Serum Albumin, Bovine; D012996:Solutions
D015306:Biogenic Monoamines; D002021:Buffers; D004800:Enzymes, Immobilized; D007202:Indicators and Reagents; D012996:Solutions; D012710:Serum Albumin, Bovine; D012701:Serotonin; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D005976:Glutaral