Novel analytical approach to a multi-sugar whole gut permeability assay

Many pathophysiological conditions are associated with increased gastrointestinal permeability, reflecting an elevated risk of endotoxaemia, inflammation, and sepsis. Permeability tests are increasingly used in clinical practice to obtain information on gastrointestinal functioning, but tests are often restricted to the small intestine, and require large oral sugar doses. Therefore, a novel multi-sugar assay was developed, allowing assessment of whole gut permeability changes in urinary and plasma samples collected at regular intervals from 10 healthy volunteers at baseline and after intake of monosaccharides (rhamnose and erythritol) and disaccharides (sucrose, lactulose, and sucralose). Samples were analyzed by isocratic cation-exchange LC-MS. Sample preparation and detection conditions were optimized. After centrifugation, chromatographic separation was achieved on an IOA-1000 column set at 30 degrees C. Column effluent was mixed with ammonia for sugar-ammonium adduct formation. The lower limit of detection was 0.05mumol/L for disaccharides and 0.1mumol/L for monosaccharides. Linearity for each probe was between 1 and 1000mumol/L (R(2): 0.9987-0.9999). Coefficients of variation were