1. A method, based on the use of the isolated frog rectus abdominis preparation, is described for studying quantitatively the hydrolysis of suxamethonium at low concentrations.2. Rates of hydrolysis of butyrylcholine, 10 mM, benzoylcholine, 0.05 mM, and suxamethonium, 0.025 mM, by plasma from different species and human plasma with genetic variants of cholinesterase were measured.3. The rates of hydrolysis of suxamethonium by different types of plasma vary widely. Human plasma with usual cholinesterase and monkey plasma hydrolyse suxamethonium more speedily than do the plasma of cats, dogs and rats, and human plasma with either atypical or fluoride resistant cholinesterase. This is only to a small extent attributable to differences in enzyme concentration and not explained by the presence of an inhibitor.4. The K(m) values for butyrylcholine are very similar for different types of plasma but the K(I) values for suxamethonium in the system plasma-butyrylcholine-suxamethonium vary greatly.5. These results and observations on the inhibition by decamethonium of the hydrolysis of butyrylcholine are consistent with the interpretation that the rates of hydrolysis of suxamethonium, 0.025 mM, obtained with different types of plasma vary because they are a function of two variables, the affinity of the ester for cholinesterase and the stability of the monosuccinyl derivative of the cholinesterase. It seems that human plasma with atypical cholinesterase hydrolyses suxamethonium much slower than does human plasma with usual cholinesterase mainly or solely because of differences in affinity of the ester for the two enzymes. On the other hand, cat plasma appears to hydrolyse suxamethonium much slower than does human plasma with usual cholinesterase mainly because the monosuccinyl derivative of cholinesterase in cat plasma is much more stable than that in human plasma. The reverse might apply for monkey plasma.6. Inhibition by dibucaine or sodium fluoride of the hydrolysis of benzoylcholine is not a general guide to rates of hydrolysis of suxamethonium by different types of plasma.