Measurements of glutathione and other thiols in cells and tissues: a simplified procedure based on the HPLC separation of monobromobimane derivatives of thiols.
Although there is intense interest in the role of thiols in controlling the efficiency of radiosensitizers, and in developing thiols (or pro-drugs liberating thiols) as radioprotectors, there is little information regarding the concentration of specific thiols in cells, tumors and normal tissues. Details are presented of a modified procedure using the thiol binding agent monobromobimane with separation using paired-ion reverse phase high performance liquid chromatography (HPLC). This method has been extended to include measurements of the radiosensitizer misonidazole and its desmethylated metabolite Ro 05-9963 in tissues.
D000818:Animals; D001643:Bridged Bicyclo Compounds; D001952:Bridged-Ring Compounds; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D005978:Glutathione / Q000032:analysis*; D007668:Kidney / Q000032:analysis; D008099:Liver / Q000032:analysis; D008624:Mercaptoethylamines / Q000032:analysis; D051379:Mice; D008920:Misonidazole / Q000031:analogs & derivatives / Q000032:analysis; D009132:Muscles / Q000032:analysis; D009374:Neoplasms, Experimental / Q000032:analysis; D011838:Radiation-Sensitizing Agents / Q000032:analysis; D013438:Sulfhydryl Compounds / Q000032:analysis*
D001643:Bridged Bicyclo Compounds; D001952:Bridged-Ring Compounds; D008624:Mercaptoethylamines; D011838:Radiation-Sensitizing Agents; D013438:Sulfhydryl Compounds; C020174:N-(2-mercaptoethyl)-1,3-diaminopropane; D008920:Misonidazole; D005978:Glutathione; C009997:desmethylmisonidazole; C028381:monobromobimane