人胃黏膜中前列腺素水平的定量测定——采用激光诱导荧光检测的微柱高效液相色谱分析法

Quantitative determination of prostaglandin levels in human gastric mucosa--analysis by microcolumn high performance liquid chromatography with laser induced fluorescence detection.

作者信息

Goto H, Sugiyama S, Kawabe Y, Kuroiwa M, Ohara A, Tsukamoto Y, Nakazawa S, Ozawa T

机构信息

Department of Internal Medicine, Faculty of Medicine, University of Nagoya, Japan.

出版信息

Biochem Int. 1990;20(6):1119-25.

PMID:2369414

Abstract

This study was designed to analyze PGs in human gastric mucosa using biopsy specimens at femtomole level by the combination of microcolumn HPLC and He/Cd laser induced fluorescence detection. Biopsy specimens were taken along the greater curvature at the corpus of the stomach, in which no gastric disease was revealed by endoscopic examination. PGs extracted from human gastric mucosa were derivatized with ADAM, and ADAM-derivatized PGs were injected into the column for analysis. The mobile phase of acetonitrile-water (73:27) containing 0.01% of phosphoric acid was used at a constant pressure of 20 kgf/cm2. Using this system, PGs in few mg of human gastric mucosa obtained by biopsy were well separated and detected; i.e., 1653 +/- 254 (femtomole/mg tissue), 279 +/- 56, 729 +/- 153, 831 +/- 199 for 6-keto-PGF1 alpha, PGF2 alpha, PGE2, and PGD2, respectively. In conclusion, the microcolumn HPLC system with laser induced fluorescence detection is a reliable method for determining individual PGs in human gastric mucosa. In addition, PGI2 is the predominant PG in human gastric mucosa and probably plays an important role in gastric function.

摘要

本研究旨在通过微柱高效液相色谱法（HPLC）与氦/镉激光诱导荧光检测相结合，在飞摩尔水平上分析人胃黏膜中的前列腺素（PGs）。在内镜检查未发现胃部疾病的胃体大弯处取活检标本。从人胃黏膜中提取的PGs用1-（二甲氨基）萘-5-磺酰氯（ADAM）进行衍生化，然后将ADAM衍生化的PGs注入柱中进行分析。使用含0.01%磷酸的乙腈-水（73:27）作为流动相，在20 kgf/cm²的恒定压力下进行分析。利用该系统，通过活检获得的几毫克人胃黏膜中的PGs得到了很好的分离和检测；即，6-酮-前列腺素F1α、前列腺素F2α、前列腺素E2和前列腺素D2的含量分别为1653±254（飞摩尔/毫克组织）、279±56、729±153和831±199。总之，带有激光诱导荧光检测的微柱HPLC系统是测定人胃黏膜中单个PGs的可靠方法。此外，前列环素（PGI2）是人胃黏膜中的主要PG，可能在胃功能中起重要作用。