Studies on intestinal permeability of cirrhotic patients by analysis lactulose and mannitol in urine with HPLC/RID/MS

Abstract

The method for separation and determination of lactulose (L) and mannitol (M) in urine was developed by HPLC with a refractive index detector (RID). The linearity ranged from 5 to 1000μg/mL for L and M, respectively. Recoveries ranged from 93.1% to 97.1%. The intra- and inter-day relative standard deviations of peak area were between 0.8–1.4% (n=3) and 1.4–3.6% (n=3). The limits of detection were obtained with 1.40μg/mL for L and 1.65μg/mL for M. The ratios of L/M in the urine samples for the spontaneous ascitic fluid infection (SAI), sterile ascitic fluid (SA) patients, and healthy volunteers (HV) were determined. The results showed well the correlations among the L/M ratio, intestinal permeability (IP) and the illness status of patients, and also indicated lactulose could improve the IP of SAI patients. The peaks of L and M in chromatograms were identified by electrospray ionization/mass spectrometry (ESI/MS), which ensured the accurate measurement of the ratio L/M. This method presented a rapid, accurate, and practical technique for determining IP in clinical practice and investigating the pathology of hepatocirrhosis.