摘要
[目的]建立通过双糖吸收试验评价儿童肠粘膜屏障功能的高效液相色谱法(high performance liquid chromatography,HPLC).[方法]采用HPLC法检测研究对象尿液标本中甘露醇(mannitol,M)和乳果糖(lactulose,L)排出率比值.以Sugar-Pak 1(300×6.5 mm,Waters)为色谱柱;柱温:85℃;流动相:超纯水;流速:0.5 ml/min;示差折射检测器.[结果]在上述色谱条件下,尿液中甘露醇和乳果糖能得到良好分离.该法检测甘露醇和乳果糖排出量的平均日内变异系数分别为2.51%和3.03%(n=4);日间变异系数分别为3.53%和5.52%;甘露醇的加样回收率在93%～98%之间,乳果糖的加样回收率在91%～97%之间.本方法测定甘露醇的最大检出限为25mg/L,乳果糖为12.5 mg/L.食物过敏儿童肠道通透性(L/M:0.18±0.06)较正常儿童(L/M:0.05±0.03)显著增高(P＜0.001).[结论]HPLC检测肠道通透性的准确性高、重复性好,是临床评价小肠粘膜屏障功能的实用方法.
[Objective] To evaluate the intestinal barrier function of children by two-sugar absorption test with a simple,specific high performance liquid chromatography( HPLC). [Methods] Urine samples were collected before and after test solution administrated, and analyzed on a Sugar-PakI column of HPLC with refractive index detector. Mobile phase:pure deionized water. Flow rate:0.5 ml/min. temprature of column:85℃. [Results] Under there conditions, the retention time of mannitol and lactulose were 13.86 min,9.27 min respectively. The average within-run and between-run reproducibilities tests were 2. 51% and 3. 53% for mannitol, 3. 03% and 5. 52% for lactulose respectively. The recovery rate raged from 93% to 98% for mannitol and 91% - 97% for lactulose. A significant rise in 5-hour urinary L/M excretion ratios was found in the children with food hypersensitivity(0. 18 ±0. 06) compared to that of control(0. 05±0. 03)(P<0. 001). [Conclusion] The determination of the L/M ratio by HPLC may be a reliable method to evaluate intestinal barrier function in clinical practice.
出处
《中国儿童保健杂志》
CAS
2005年第1期29-32,共4页
Chinese Journal of Child Health Care
关键词
肠道通透性
高效液相色谱法
乳果糖
甘露醇
儿童
intestinal permeability
high performance liquid chromatography
lactulose
mannitol
child
