title: Metabolic toxicity screening using electrochemiluminescence arrays coupled with enzyme-DNA biocolloid reactors and liquid chromatography-mass spectrometry.
abstract: New chemicals or drugs must be guaranteed safe before they can be marketed. Despite widespread use of bioassay panels for toxicity prediction, products that are toxic to a subset of the population often are not identified until clinical trials. This article reviews new array methodologies based on enzyme/DNA films that form and identify DNA-reactive metabolites that are indicators of potentially genotoxic species. This molecularly based methodology is designed in a rapid screening array that utilizes electrochemiluminescence (ECL) to detect metabolite-DNA reactions, as well as biocolloid reactors that provide the DNA adducts and metabolites for liquid chromatography-mass spectrometry (LC-MS) analysis. ECL arrays provide rapid toxicity screening, and the biocolloid reactor LC-MS approach provides a valuable follow-up on structure, identification, and formation rates of DNA adducts for toxicity hits from the ECL array screening. Specific examples using this strategy are discussed. Integration of high-throughput versions of these toxicity-screening methods with existing drug toxicity bioassays should allow for better human toxicity prediction as well as more informed decision making regarding new chemical and drug candidates.
authors.affiliation: Department of Chemistry, East Carolina University, Greenville, North Carolina 27858, USA. hvastkovse@ecu.edu
authors.forename: Eli G
authors.initials: EG
authors.lastname: Hvastkovs
authors.forename: John B
authors.initials: JB
authors.lastname: Schenkman
authors.forename: James F
authors.initials: JF
authors.lastname: Rusling
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D004247:DNA / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D018736:DNA Adducts / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000378:metabolism; D004867:Equipment Design; D006801:Humans; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods; D008958:Models, Molecular; D009152:Mutagenicity Tests / Q000295:instrumentation* / Q000379:methods*; D009153:Mutagens / Q000378:metabolism / Q000633:toxicity*
chemical_list: D018736:DNA Adducts; D004800:Enzymes, Immobilized; D009153:Mutagens; D004247:DNA