气相色谱法同时测定鼠李糖、乳果糖和蔗糖及其在胃肠道通透性检测中的应用

Simultaneous gas-chromatographic measurement of rhamnose, lactulose and sucrose and their application in the testing gastrointestinal permeability.

作者信息

Abazia Cristiana, Ferrara Renata, Corsaro Maria Michela, Barone Gaspare, Coccoli Pietro, Parrilli Gianpaolo

机构信息

Dipartimento di Medicina Clinica e Sperimentale, Università Federico II,Via S. Pansini, 5 80131 Naples, Italy.

出版信息

Clin Chim Acta. 2003 Dec;338(1-2):25-32. doi: 10.1016/j.cccn.2003.07.018.

DOI:10.1016/j.cccn.2003.07.018

PMID:14637262

Abstract

BACKGROUND

As it is important to test gastric and intestinal permeability simultaneously in gastrointestinal disorders such as Celiac disease, we developed a gas-chromatographic (GC) method to estimate rhamnose (L-rh), lactulose (Lacl) and sucrose (Suc) in urine.

METHODS

The method is based on the use of alditol acetate derivatives giving a lower number of GC peaks than reducing sugars do. Acetate derivatives are more stable and less expensive than GC silylates and keep the flame-detector cleaner. We checked the chemical stability of alditol acetates by verifying the reproducibility of the standard curve of a sugar derivative sample which had been stored for 2 months at -20 degrees C.

RESULTS

The calibration proved linear over the range 0.1-1 microg of sugar injected. Analytical sugar recovery was 88%+/-19.4% (mean+/-S.D.) for rhamnose, 105%+/-7.4% for sucrose and 102%+/-2.4% for lactulose. Mean within-day precision (CV) was 7.7% for rhamnose, 5.7% for sucrose and 1.9% for lactulose, and between-day (CV) was 6.7% for rhamnose, 3.9% for sucrose and 1.6% for lactulose. The rhamnose, lactulose and sucrose as the lactulose/rhamnose ratio clearly differentiated 25 healthy controls from 36 patients with active gluten-sensitive enteropathy.

CONCLUSIONS

A fast, reliable and cheap gas-chromatographic method is presented here to evaluate gastric and intestinal permeability.

摘要

背景

由于在诸如乳糜泻等胃肠道疾病中同时检测胃和肠道通透性很重要，我们开发了一种气相色谱（GC）方法来测定尿液中的鼠李糖（L-rh）、乳果糖（Lacl）和蔗糖（Suc）。

方法

该方法基于使用乙酸糖醇衍生物，其产生的GC峰数量比还原糖少。乙酸衍生物比GC硅烷化试剂更稳定、更便宜，并且能使火焰检测器更清洁。我们通过验证在-20℃下储存2个月的糖衍生物样品标准曲线的重现性来检查乙酸糖醇的化学稳定性。

结果

在所注入糖的0.1 - 1微克范围内，校准证明呈线性。鼠李糖的分析糖回收率为88%±19.4%（平均值±标准差），蔗糖为105%±7.4%，乳果糖为102%±2.4%。鼠李糖的日内精密度（CV）平均值为7.7%，蔗糖为5.7%，乳果糖为1.9%；日间精密度（CV）鼠李糖为6.7%，蔗糖为3.9%，乳果糖为1.6%。以乳果糖/鼠李糖比值表示的鼠李糖、乳果糖和蔗糖能清楚地区分25名健康对照者和36名活动性麸质敏感肠病患者。