Establishment of the method of detection of intestinal permeability in mouse model of total body irradiation using high-performance liquid chromatography evaporative light-scattering detector

Objective:To establish the method of employing high-performance liquid chromatog-raphy evaporative light-scattering detector(HPLC-ELSD) using two-sugar absorption test for evalu-ating the intestinal permeability in mouse model total body irradiation(TBI).Methods:Eight urine smaples of the total body irradiation(TBI) and the healthy control were collected respectively to evaluate the intestinal permeability in two groups mice for simultaneous determination of urinary mannitol/lactulose(L/M) excretion ratios.Results:The calibration curve of mannitol and lactulose were linear in the concentration range of 0.5～6 mg/mL and 0.25～3 mg/mL.The LOD of mannitol and lactulose was 500 ng and 250 ng,respectively.The recovery rate ranged from 86.5%～109.2% for mannitol and 86.3%～114.4% for lactulose.The average within-run and between-run reproducibilities tests were 0.86% and 1.13%(n =6)for mannitol,1.11% and 1.39%(n =6)for lactulose,respectively.A significant rise in 6 hour urinary L/M excretion ratios was found in the TBI mice(0.5108±0.03266) compared to that of control(0.2908±0.05332)(P 0.05).Conclusion:The determination of the L/ M ratio by HPLC is simple,specific,sensitive and reproducible.It may be a reliable method to evaluate and monitor intestinal permeability in the TBI mice.