Fluorometric Detection and etermination of Carbohydrates in Thin-layer Chromatography and High-Performance Liquid Chromatography Using Taurine-Borate Reagent

Abstract

Highly sensitive fluorometric detection and determination of carbohydrates in thin-layer chromatography (TLC) and iiigh-performance liquid chromatography (HPLC) have been developed using, t aurine-borate reagent- which gives intense fluorescence on reaction with carb ohydrates by heating in neutral or slightly alkaline media.,
The flnorescence development on thin-layer plate was performed by spraying an 8% taurine solution in 0.05 m ol. d m^-3 Na₂B₄O₇-0.1m oldm^-3 KH₂PO₄ buffer, pH 7.5, on a thin -layer plate which was then heated at 130°C for 10 m in and by measuring the fluorescence intensity with a scaning, fluorophotometer. Detection limits for reducing sugars were in th e range between 0.1 nmol and 0.2 nmol. Standard curves for glucose and galactose were linear i n the range from 0.05 μg/spott o 1.0μg/spot and for lactose from 0.1 μg/spotto 1.0 μg/spot.
HPLC was carried out by eluting a sample solution through an anion Hexchange Aminex A-27 column or a reversed phase Lichrosorb-NH₂ column, heating the eluate after mixing with an 8% taurine solution or an 8% taurine solution containing 0.05 m ol. dm^-3 borate adjusted to pH 8.7 with NaOH pellets and measuring the fluorescence intensity with a fluor omonitor. The present method facilitated excellent separation of mono- and oligosaccha ride mixtures within a remarkably short period. Since this reaction proceeds without the use of str ong acid or alkali, the chromatographic methods do not require acid or alkali resistant equipm ents.