HPLC detection of soluble carbohydrates involved in mannitol and trehalosemetabolism in the edible mushroom Agaricus bisporus

A convenient and sensitive method was developed to separate and detect vari ous types of carbohydrates (polyols, mono- and disaccharides, and phosphory lated sugars) simultaneously using high-performance liquid chromatography ( HPLC). The method consists of a chromatographic separation on a CarboPac PA 1 anion-exchange analytical column followed by pulsed amperometric detectio n. In a single run (43 min) 13 carbohydrates were readily resolved. Calibra tion plots were linear over the ranges of 5-25 mu M to 1.0-1.5 mM. The reli able and fast analysis technique, avoiding derivatization steps and long ru n times, was used to determine the levels of carbohydrates involved in mann itol and trehalose metabolism in the edible mushroom Agaricus bisporus. Mor eover, the method was used to study the trehalose phosphorylase reaction.