高压液相色谱法检测梗阻性黄疸大鼠肠道通透性  

Detection of Intestinal Permeability of the Rats with Obstructed Jaundice by High Performance Liquid Chromatographic Method

作　　者：曹萌[1] 时开网[1] 何桂珍[2] 倪永军[1] 宋毕清[1] 

机构地区：[1]南京医科大学附属南京第一医院普通外科,南京210006 [2]北京协和医院营养科,北京100000

出　　处：《现代检验医学杂志》2012年第2期26-29,共4页Journal of Modern Laboratory Medicine

基　　金：南京市重点课题

摘　　要：目的用高压液相色谱方法快速准确测量肠道通透性，探讨正常和阻塞性黄疸大鼠的肠道通透性状态。方法24只SD大鼠被随机平均分成2组：健康对照组和梗阻性黄疸组。胆总管完全结扎后获得梗阻性黄疸动物模型。测定所有SD大鼠第6天尿中乳果糖和甘露醇溶液的排出率。所有SD大鼠在第5天晚10时起禁食；第6天早8时灌胃2ml乳果糖和甘露醇溶液（含100mg乳果糖和50mg甘露醇），收集之后24h内的全部尿液。尿样经离心和预处理后，用带脉冲电化学检测器的高压液相离子色谱仪测定，样本分析时间为10min。结果甘露醇和乳果糖标准曲线线性良好，线性的相关系数r分别为0．998823和0．999943；方法学的重复性误差（CV％，，n=12）：乳果糖（I。）3．52，甘露醇（M）1．61；回收率：乳果糖（L）98％．4±3％，甘露醇（M）101．1％±3％。第1组和第11组尿中的乳果糖（L）和甘露醇（M）排出率（％）的比值（L／M）分别为0．33土0．04和2．OO土0．16，第1I组l，／M值显著比第1组大（P〈O．001）。结论带脉冲电化学检测器的高压液相离子色谱仪测定尿中乳果糖和甘露醇的排出率，方法准确、重复性和回收率满意。梗阻性黄疸SD大鼠L／M比值的测定可反映肠黏膜的通透性和肠道屏障功能。Objective To detect the intestinal permeability quickly and accurately by high performance liquid chromatography （HPI.C） and to study the intestinal permeability of rats associated with the normal and obstructive jaundice. Methods 24 Sprague-Dawley rats were randomly assigned into 2 groups of 12 each： Control group （group Ⅰ）and obstructive jaundice group（groupⅡ）. The common bile duct was ligated completely to obtain obstructive jaundice model. All subjects had to fast from 10 ： 00 pm in 5th day and were administrated by gavage with 2 ml of lactulose and mannitol solution （containing100 mg lactulose and 50mg mannitol）at 8：00 am in 6th day. All urine of subjects were collected within the following 2,4 h. After urine were centrifuged and pretreatmented, the samples were measured by HPLC with pulsed electrochemical detector. A run analysis time was 10 mira Results Mannitol and lactulose standard curve were good linearity and r were 0.99g 823 and 0. 999 943 respectively. The repeatability error （CV%, n= 12） of lactulose （L） and mannitol（M） were 3.52 and 1.61 respec- tively. The rate of recovery were lactulose（L） 98.4%±3% and mannitol（M） 101.1%±3% The rate of lactulose and man- nitol（L/M） in Group I and Group Ⅱ were 0. 33±0. 04 and 2.00±0.16 respectively,which in group lI was significantly higher than it in group I （P〈0. 001）. Conclusiun Detection of lactulose and mannitol by the high-performance liquid ion chromatography with pulsed electrochemical detector is accurate. That reproducible and recovery are satisfactory. The ratio of excretion rate in rats with obstructive jaundice can reflect the permeability and mucosa barrier function of intestinal.

关 键 词：高压液相色谱 乳果糖/甘露醇 肠道通透性 

分 类 号：R332[医药卫生—人体生理学]