Evaluation of chrono-amperometric signal detection for the analysis of genotoxicity by a whole cell biosensor.
Electrochemical signal detection can be readily integrated in biosensors and is thus an attractive alternative to optical detection methods. In the field of environmental chemistry and ecotoxicology there is a growing demand for lab-independent devices based on whole cell biosensors for the detection of genotoxic compounds. Because of the broad occurrence of pre-genotoxic compounds that need to be bio-activated, the integration of a system for metabolic activation into such a biosensor is important. The present study evaluates a chrono-amperometric detection method in which para-aminophenyl beta-D-galactopyranoside is used as substrate for a reporter gene assay based on the bacterial SOS-response in comparison to a test system for the determination of genotoxicity in water that is standardized according to the International Organization for Standardization (ISO). The evaluation was done in order to analyze the potential of the electrochemical signal detection to be used as a complementary method for the standard test system and thus to evaluate the usability of electrochemical biosensors for the assessment of genotoxicity of environmental samples. In the present study it is shown that the chrono-amperometric detection of para-aminophenol is specific even in the presence of electro-active species generated by the enzymatic system used for the external bio-activation of contaminants. Under optimized conditions electrochemistry is sufficiently sensitive with a limit of detection that is comparable to the respective ISO-standard.
Buchinger
Sebastian
S
Division of Qualitative Hydrology, German Federal Institute of Hydrology (BfG), Am Mainzer Tor 1, Koblenz 56068, Germany. Buchinger@bafg.de
Grill
Pia
P
Morosow
Valeri
V
Ben-Yoav
Hadar
H
Shacham-Diamand
Yosi
Y
Biran
Alva
A
Pedahzur
Rami
R
Belkin
Shimshon
S
Reifferscheid
Georg
G
D015112:4-Nitroquinoline-1-oxide / Q000032:analysis / Q000633:toxicity; D015374:Biosensing Techniques / Q000379:methods*; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D004784:Environmental Monitoring; D005697:Galactosides / Q000737:chemistry*; D017930:Genes, Reporter; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity*; D011804:Quinolines / Q000032:analysis / Q000633:toxicity; D013014:SOS Response, Genetics / Q000187:drug effects; D012486:Salmonella typhimurium / Q000187:drug effects / Q000201:enzymology; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D005697:Galactosides; D009153:Mutagens; D011804:Quinolines; C029216:2-amino-3-methylimidazo(4,5-f)quinoline; C010677:4-aminophenyl-beta-galactoside; D015112:4-Nitroquinoline-1-oxide; D001616:beta-Galactosidase