Abstract
A single-stage post-column fluorescence HPLC method for the analysis of N-methylcarbamates and N-methylcarbamoyloximes is presented as an alternative to the traditional two-stage method. The two-stage technique involves post-column hydrolysis with sodium hydroxide followed by derivatization with o-phthalaldehyde and β-mercaptoethanol in a borate buffer. In the proposed method only one reagent, consisting of sodium hydroxide, o-phthalaldehyde, and N,N-dimethyl-2-mercaptoethylamine hydrochloride (Thiofluor), is used. The stability of the reagent is compared to single-stage alternatives involving β-mercaptoethanol or 3-mercaptopropionic acid, which have been previously reported. It is reported that Thiofluor provides the best stability and is satisfactory for overnight chromatographic runs. The sensitivity of the method is reported to be equal to that of the two-stage method. This innovation simplifies reagent preparation and equipment maintenance, while shortening start-up time without any loss in sensitivity.
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