Specific Radioisotopic Assay for Cholinesterase
Technical rept. Dec 1987-Mar 1988
ARMY MEDICAL RESEARCH INST OF CHEMICAL DEFENSE ABERDEEN PROVING GROUND MD
Pagination or Media Count:
The radiometric method I for measuring ChE activity Siakotos et al., Biochem. Med. 3, 1, 1969 was modified to preclude the use of p-dioxane, a hazardous material. The modified procedure II uses 0.4 N perchloric acid PCA , instead of p-dioxane, to denature the ChE and stop hydrolysis of 14C- acetylcholine ACh. The unreacted substrate ACh is removed by cationic exchange resin suspended in water. The supernatant acidic water solution containing the product of hydrolysis, 14C-acetic acid, is mixed with nonhazardous scintillation cocktail and counted. The incubation mixture 37 degrees C for II is similar to I and is composed of 0.1 ml of buffer-salt solution pH 7.8, 0.1 ml of guinea pig whole blood WB-water suspension and 0. 1 ml of 3mM ACh solution. Keywords Acetylcholinesterase Acetylcholine Radiometric ChE assay Physostigmine Irreversible ChE inhibitors Titrametric ChE assay.
- Test Facilities, Equipment and Methods
- Recording and Playback Devices