A biological method for the quantitative measurement of tetrodotoxin (TTX): tissue culture bioassay in combination with a water-soluble tetrazolium salt.
A tissue culture bioassay, using the mouse neuroblastoma cell line (Neuro2A), was improved to provide a simple and sensitive bioassay for TTX or sodium channel-blocking toxins (SCB). The water-soluble tetrazolium salt, 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetraz olium, monosodium salt (WST-1), was applied to replace the time-consuming and subjective cell-counting procedure of the cells with automatic measurement, using a microplate reader. It was also confirmed that this method is directly applicable to bacterial culture supernatants, with the precaution of possible interference.
Hamasaki
K
K
Department of Bioengineering, Faculty of Engineering, Soka University.
Kogure
K
K
Ohwada
K
K
D000818:Animals; D001681:Biological Assay; D046508:Culture Techniques; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D051379:Mice; D009447:Neuroblastoma / Q000473:pathology; D015203:Reproducibility of Results; D012995:Solubility; D013778:Tetrazolium Salts / Q000737:chemistry*; D013779:Tetrodotoxin / Q000032:analysis* / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014867:Water
D013778:Tetrazolium Salts; D014867:Water; D013779:Tetrodotoxin