Abstract
In this paper, a novel sensor platform based on screen printed carbon electrode coated by graphene modified polyacrylamide gel (GR/PAAGC) was developed and implemented for sampling, detection and enumeration of coliform bacteria (coliforms) on food contact surfaces. The optimized formula of polyacrylamide (PAA) and agar-agar increased the adhesive properties of the gel, being crucial for the coliforms recovery, attached to food contact surfaces. The 6-Chloro-3-indoxyl-β-D-galactopyranoside (6-CIGP) was used as a new electrochemical reporter for β-D-galactosidase activity. The released 6,6′-Dichloro-Indigo (6-DI) was directly detected by GR/PAAGC sensor. The presence of Isopropyl-β-D-thiogalactopyranoside (IPTG) and n-Octyl-β-D-thiogalactopyranoside (OBDG) in the gel contributed to reduction of the detection time. The addition of graphene enhanced the voltammetric signal and increased the conductivity of PAA gel. The anodic and cathodic peaks of the released product were directly proportional to the concentration of coliforms. Bacterial cell concentrations ranging from 1.6log10CFU/mL to 6.6log10CFU/mL were detected. Well-shaped, sharp voltammetric curves were generated within 3 h. Redox peaks exhibited good sensitivity with detection limits (LOD) < 0.6log10CFU/mL. After series of optimization experiments, coliforms ranging from 0.6log10CFU/cm2 to 6.610CFU/cm2 on stainless steel surfaces have been detected within 30 min with a LOD of 0.1log10CFU/cm2. The developed rapid, sensitive, reproducible and specific sensor successfully applied for single detection as well as for real-time monitoring of growth of coliform bacteria on stainless steel surfaces during food processing.
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