Chromatographic analysis of sugars in physiological fluids by postcolumn reaction with cuprammonium: a new and highly sensitive method.

A new highly sensitive method of high-performance liquid chromatographic analysis has been used to separate mono-, di-, and oligosaccharides present in jejunal aspirates from experimentally perfused animals. The sugars in the column eluant are detected at 280 to 310 nm after reaction with cuprammonium. No reaction coil is required between the column and uv detector because the reaction is virtually instantaneous. This technique is more sensitive than differential refractometry, so that it is possible to detect 2.5 nmol (450 ng) of glucose routinely, compared to a minimum detection limit of 100 nmol (18 micrograms) using a commercial refractive index detector. Using a suitable postcolumn pump, the technique is not sensitive to changes in solvent composition, as with gradient elution, and is applicable to analysis of other cis-cis glycols.