On-column catalysis of hydratization equilibria during liquid chromatographic analysis of acetaldehyde and formaldehyde

Summary The aldehydes were separated by reversed phase chromatography on a polymer-based support. Acid catalysis on the column was needed to obtain sufficiently fast equilibrium between acetaldehyde and its hydrate. Deuterium labeled acetaldehyde (D₃) yielded complete separation from the nonlabeled compound, probably due to a difference in hydratization. Fluorescent products were obtained by reaction between the aldehydes and a mixture of ammonia and dimedone (5.5-dimethyl-1.3-cyclohexanedione). The fluorescence was detected at 460 nm after excitation at 390 nm. A reaction time of 42 seconds at 90°C in a knitted teflon capillary produced detection limits of 0.5 and 1 ng for acetaldehyde and formeldehyde, respectively. A method for preparation of a stable reagent with low background fluorescence is described.