Modern approach for determination of lactulose, mannitol and sucrose in human urine using HPLC–MS/MS for the studies of intestinal and upper digestive tract permeability

► Simultaneous analysis of mannitol, lactulose and sucrose. ► Rapid sample preparation. ► Estimation of upper digestive tract and intestinal permeability at single analysis. ► Comparison of three analytical columns. A new analytical procedure was described for the simultaneous determination of lactulose, mannitol and sucrose in urine, in which HILIC chromatography and tandem mass spectrometry detection are used. Sugars are orally administered for the estimation of intestinal permeability in children digestive tract. Samples were purified by dispersive solid phase extraction (d-SPE) using Amberlite MB150 resin. Raffinose was selected as an internal standard. The chosen chromatographic separation was carried out on ZIC®-HILIC column in 10min at a flow rate of 0.3mL/min, using mixture of acetonitrile (ACN) and ammonium acetate (NH4Ac) in water (H2O) as the mobile phase. Within-run precision (CV) measured at three concentrations was 1.08%, 0.32% and 0.49% for lactulose; 1.88%, 0.47% and 0.75% for mannitol, 2.95%, 1.31% and 0.6% for sucrose. Between-run CVs were 0.75%, 1.1% and 1.2% for lactulose; 1.1%, 1.02% and 1.01% for mannitol; 1.17%, 1.4% and 1.05% for sucrose. Analytical recovery of all three sugar probes was 95.06–99.92%. The detection limits were: 15.94ng/mL for lactulose, 17.10ng/mL for sucrose and 11.48ng/mL for mannitol. The proposed method is rapid, simple, sensitive and suitable for the determination of intestinal permeability of the sugar derivatives in children.