高效液相色谱用于肝硬化大鼠肠道通透性的测定  

Detection of intestinal permeability in cirrhotic rats using HPLC

作　　者：覃慧敏[1] 吴春明[1] 李洪涛[1] 李国军[1] 宋建新[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院感染科

出　　处：《华中医学杂志》2008年第2期113-115,共3页Central China Medical Journal

基　　金：湖北省卫生厅科研基金课题(NX200506)

摘　　要：目的建立通过双糖吸收试验评价肝硬化大鼠肠道通透性的高效液相色谱法(HPLC)。方法建立四氯化碳诱导大鼠肝硬化模型。采用HPLC检测30例正常大鼠和30例肝硬化大鼠尿液标本中甘露醇和乳果糖排出率比值。以NH2柱(4.6mm×250mm×5mm,Warters)为色谱柱;柱温为室温;流动相为乙腈∶水70∶30(V/V);流速为1.0ml/min;进样量为20μl。示差折光检测器(RID2000)。结果在上述色谱条件下,尿样中甘露醇和乳果糖能得到良好基线分离,甘露醇和乳果糖的出峰时间分别为6.721min和9.242min。该法检测甘露醇和乳果糖的线性范围分别为5～1000μg/ml、2.5～500μg/ml。甘露醇的加样回收率在92.7%～98.0%,乳果糖的加样回收率在91.3%～97.4%。肝硬化大鼠肠道通透性(乳果糖/甘露醇比值:0.03578±0.01245)显著高于正常大鼠(0.02563±0.00871)。结论HPLC检测肠道通透性具有准确性高、重复性好、简便灵敏等特点,可用于肝硬化大鼠肠道通透性的测定和监测。Objective To evaluate the intestinal permeability in cirrhotic rats by using two-sugar absorption test with a high performance liquid chromatography（HPLC）. Methods Liver-cirrhosis model in rats was induced by carbon tetrachloride. Thirty urine samples of the cirrhotic rats and the healthy control were collected respectively after test solution administrated, and analyzed on a NH2 column of HPLC with refractive index detector2000. Temperature of column： ambient temperature. Mobile phase： CH3CN/H20 （70/30, V/V）. Flow rate： 1.0 ml/miru Injection volume： 20 /11. Results Under these conditions, the retention time of mannitol and lactulose were 6. 271 min, 9. 242 min respectively. The recovery rate raged from 92. 7% to 98. 0K for mannitol and 91.3%～97. 4% for lactulose. A significant rise in 8 hour urinary L/M excretion ratios was found in the cirrhotic rats（0. 03578 ± 0. 01245） compared to that of control（0. 02563 ± 0. 00871） （P〈0. 05）. Conclusion The method of detection of the L/M ratio by HPLC is rapid, sensitive,accurate and reproducible and may be a reliable method to evaluate and monitor intestinal permeability in cirrhotic rats.

关 键 词：高效液相色谱 肝硬化 肠道通透性 乳果糖 甘露醇 

分 类 号：R575.2[医药卫生—消化系统]