高效液相色谱法分离和定量无菌大鼠粪便及肠内容物中的胆红素及其葡萄糖醛酸结合物。

HPLC separation and quantification of bilirubin and its glucuronide conjugates in faeces and intestinal contents of germ-free rats.

作者信息

Saxerholt H, Skar V, Midtvedt T

机构信息

Department of Medical Microbial Ecology, Karolinska Institute, Stockholm, Sweden.

出版信息

Scand J Clin Lab Invest. 1990 Sep;50(5):487-95. doi: 10.1080/00365519009089163.

DOI:10.1080/00365519009089163

PMID:2237261

Abstract

We describe an accurate reverse-phase high-performance liquid chromatography (HPLC) method for the separation and quantification of unconjugated bilirubin (UCB) and its monoglucuronide (BMG) and diglucuronide (BDG) conjugates in faeces and intestinal contents from germ-free (GF) rats. We demonstrated that female GF rats excreted predominantly BMG and that the percentage of this conjugate was at most 71.7% of the total bilirubin excreted with the faeces. The highest percentages for BDG and the UCB were 27.9% and 6.0%, respectively. The bile pigment composition in duodenal contents was 59.8% BDG and 40.2% BMG (median percentage) and was 47.7% BDG, 50.1% BMG and 2.2% UCB in ileal contents. Deconjugation of BDG to BMG was profound in caecal contents with 26.0% BDG, 67.4% BMG and 6.6% UCB. Endogenous (mammalian) beta-glucuronidase activity was present in intestinal contents throughout the entire length of the intestine and in faeces of the GF rats. The results indicated that it is very likely that endogenous beta-glucuronidase plays a role in the deconjugation of bilirubin glucuronides as well as of other glucuronides in the intestine of the GF rat.

摘要

我们描述了一种准确的反相高效液相色谱（HPLC）方法，用于分离和定量无菌（GF）大鼠粪便和肠内容物中未结合胆红素（UCB）及其单葡萄糖醛酸苷（BMG）和双葡萄糖醛酸苷（BDG）共轭物。我们证明，雌性GF大鼠主要排泄BMG，该共轭物的百分比最高占随粪便排泄的总胆红素的71.7%。BDG和UCB的最高百分比分别为27.9%和6.0%。十二指肠内容物中的胆汁色素组成是59.8% BDG和40.2% BMG（中位数百分比），回肠内容物中是47.7% BDG、50.1% BMG和2.2% UCB。在盲肠内容物中，BDG向BMG的去共轭作用显著，其中BDG占26.0%，BMG占67.4%，UCB占6.6%。在GF大鼠整个肠道的肠内容物和粪便中均存在内源性（哺乳动物）β-葡萄糖醛酸酶活性。结果表明，内源性β-葡萄糖醛酸酶很可能在GF大鼠肠道中胆红素葡萄糖醛酸苷以及其他葡萄糖醛酸苷的去共轭作用中发挥作用。