title: A stable in vitro method for assessing the toxicity of potassium cyanide and its antidote.
abstract: BACKGROUND
  Hydrogen cyanide possesses a high acid-dissociation constant of 9.14, favoring its vaporization and depletion from the culture media at physiological pH, which may cause the cyanide toxicity unstable in vitro.
  
  
  OBJECTIVE
  We investigated whether adjustment of culture medium pH stabilizes cyanide concentration and decreases the effective concentration of potassium cyanide (KCN).
  
  
  METHODS
  Murine fibroblast cells were exposed to different concentrations of KCN in media maintained at pH 7.4 or 9.2, in the presence or absence of hydroxocobalamin. After incubation for 1 h, we evaluated medium pH, cyanide concentration, cytochrome activity, and cell viability.
  
  
  RESULTS
  Cyanide concentration decreased to 18.8% in pH 7.4 medium compared to 83.2% in pH 9.2 medium. A significant decrease in cytochrome activity was observed at 40 mM and 1.25 mM KCN in pH 7.4 and pH 9.2 media, respectively. In pH 9.2 medium, dose-dependent cytotoxicity of KCN and antidotal effects of hydroxocobalamin were observed.
  
  
  CONCLUSIONS
  Adjustment of culture medium pH to 9.2 could stabilize cyanide concentration and decrease the effective concentration of KCN, allowing stable evaluation of KCN toxicity and antidotal efficacy.
authors: Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan. yamagiwa@is.icc.u-tokai.ac.jp.
authors: Takeshi
authors: T
authors: Yamagiwa
authors: Sadaki
authors: S
authors: Inokuchi
authors: Takeshi
authors: T
authors: Saito
authors: Shigeaki
authors: S
authors: Inoue
authors: Seiji
authors: S
authors: Morita
authors: Akira T
authors: AT
authors: Kawaguchi
mesh_terms: D000818:Animals; D000931:Antidotes / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects*; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry*; D003580:Cytochromes / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D006879:Hydroxocobalamin / Q000494:pharmacology*; D051379:Mice; D011190:Potassium Cyanide / Q000032:analysis* / Q000633:toxicity*; D018675:Toxicity Tests / Q000379:methods*; D014835:Volatilization
chemical_list: D000931:Antidotes; D003470:Culture Media; D003580:Cytochromes; D011190:Potassium Cyanide; D006879:Hydroxocobalamin