全身照射小鼠模型肠道通透性HPLC-ELSD检测方法的建立

目的：建立评价全身照射（total body irradiation,TBI）实验小鼠模型肠黏膜通透性双糖吸收试验的高效液相色谱（high-performance liquid chromatography,HPLC）检测方法。方法：TBI实验小鼠模型和正常小鼠各8只,采用高效液相色谱-蒸发光散射检测器分析（HPLC-ELSD）检测尿液标本中乳果糖（lactulose,L）和甘露醇（mannitol,M）的排出率比值,评价两组小鼠的肠道黏膜通透性。结果：尿液中甘露醇和乳果糖浓度分别在0.5～6 mg/mL和0.25～3 mg/mL范围内线性关系良好,本方法测定甘露醇和乳果糖的最低检测极限分别为500 ng和为250 ng。甘露醇和乳果糖的加样回收率分别为86.5%～109.2%和86.3%～114.4%。甘露醇和乳果糖排出量的平均日内变异系数分别为0.86%和1.13%（n=6）;平均日间变异系数分别为1.11%和1.39%（n=6）。TBI组小鼠肠道通透性（乳果糖/甘露醇比值：0.5108±0.03266）显著高于正常组小鼠（0.2908±0.05332）。结论：HPLC-ELSD检测肠道通透性具有简便、专一灵敏、重复性好、准确可靠等特点,可用于TBI小鼠肠道黏膜通透性的测定和监测。

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.