A novel HPLC method for the simultaneous quantification of monosaccharides and disaccharides used in tests of intestinal function and permeability

Abstract

The quantification of monosaccharides and disaccharides used as probes in intestinal function and permeability tests can be technically demanding, detracting from the value of this approach to the indirect assessment of intestinal damage. In this study, a procedure is described for the simultaneous quantification of rhamnose, lactulose, 3-O- methyl- d-glucose and xylose in urine by HPLC using an anion exchange column with pulsed amperometric detection. This method is relatively fast and simple to perform, requiring no pre-treatment of urine samples or post-column derivatization. Accuracy and precision of determinations are illustrated by analytical recoveries (mean percentage ± S.D., C.V., n = 30) for multiple batch analyses of a diluted urine sample containing 20 mg/l of rhamnose (100 ± 6.8, 6.2%), lactulose (100 ±6.1, 5.5%), 3-O- methyl- d-glucose (98 ± 5.9, 5.5%) and xylose (104 ± 7.1, 6.5%). Linearity of standard curves indicated that the lower limit for accurate quantification was 0.1mg/l for all four sugars. Urinary recoveries following oral administration of these sugars to control dogs were determined as a baseline for the investigation of intestinal damage in this species and comparison of chromatograms illustrated enhanced permeability in dogs with gluten-sensitive enteropathy.