New fluorometric method for estimation of citrovorum factor

Abstract

A rapid method for the identification and estimation of CF is described. After separation on paper chromatograms, CF gives a grayish blue fluorescence under ultraviolet light on reaction with acidified orcinol. The compound formed is eluted with sodium bicarbonate solution and the fluorescence is quantitatively measured in a Klett fluorometer. This method has been used to estimate enzymically formed CF in an in vitro system of chicken liver homogenate. The possibility of developing the fluorescence in solution has also been demonstrated. Even 100 mµg of CF is detectable visually on paper chromatograms. In solution, 0.5–5.0 µg of CF can be assayed with Klett fluorometer.