Solid hydroxylammonium sulfate is reacted with an alcohol solution of an alkali metal hydroxide or alkoxide to produce an alcoholic hydroxylamine liquid phase and a sulfate-containing solid phase. The liquid phase may be used for further reactions such as oxidations, hydroxamic acid production or neutralization to other hydroxylammonium salts. Tollens' reagent is a chemical reagent used to determine the presence of aldehyde, aromatic aldehyde, and alpha-hydroxy ketone functional groups. The reagent consists of a solution of silver nitrate, ammonia and some sodium hydroxide (to maintain a basic pH of the reagent solution).

The efficiency of reaction depended on the hydroxylamine concentration, denaturant, pH, and the fused protein concentration. The optimal cleavage solution consisted of guanidine ⋅ HCl as the denaturant, pH 8.1, and 6.7 mg ml −1 of fused MSI-344.