Determination of Rhein Anthrone in Caecal Content of Rats Using Reversed Phase HPLC

Abstract

A sensitive spectrofluorimetric method is described to determine small quantities of rhein anthrone in complex biological systems as caecum content and faeces. The method is based on the stability and fluorescent properties of anthrone molecules in borax solution and combines the sensitivity of spectrofluorimetric methods with the resolution power of liquid chromatography.

After extraction of the faeces with a solution of sodium tetraborate and ascorbic acid in water and centrifugation, an aliquot of the supernatant is injected on a Sephadex-G25 precolumn. After rejection of the first fractions (fluorophors present in caecal content), rhein anthrone is collected on an RP-C18 column and eluted with an acetonitrile-borax buffer gradient. The anthrone is quantified using a fluorescence HPLC monitor.