一种用于测定脑中多磷酸肌醇的高效液相色谱法。

A high-performance liquid chromatographic method for the determination of polyphosphoinositides in brain.

作者信息

Nakamura T, Hatori Y, Yamada K, Ikeda M, Yuzuriha T

机构信息

Tsukuba Research Laboratories, Eisai Company, Ltd., Ibaraki, Japan.

出版信息

Anal Biochem. 1989 May 15;179(1):127-30. doi: 10.1016/0003-2697(89)90212-1.

DOI:10.1016/0003-2697(89)90212-1

PMID:2547321

Abstract

A simple high-performance liquid chromatographic method for the determination of endogenous phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2) in brain has been developed. PIP and PIP2 were derivatized with 9-anthryldiazomethane to yield (9-anthryl)PIP and di(9-anthryl)-PIP2. The derivatives were separated on a reversed-phase column using isocratic elution and detected with a uv detector. The detection limits of PIP and PIP2 were 0.25 micrograms. The method with uv detection was sufficiently sensitive to measure the concentrations of PIP and PIP2 in rat brain. The levels of PIP and PIP2 were increased in developing rat brain and were decreased after 10 min of ischemia.

摘要

已开发出一种简单的高效液相色谱法，用于测定脑内源性磷脂酰肌醇4-磷酸（PIP）和磷脂酰肌醇4,5-二磷酸（PIP2）。PIP和PIP2用9-蒽重氮甲烷衍生化，生成（9-蒽基）PIP和二（9-蒽基）-PIP2。衍生物在反相柱上采用等度洗脱进行分离，并用紫外检测器检测。PIP和PIP2的检测限为0.25微克。紫外检测法灵敏度足以测量大鼠脑内PIP和PIP2的浓度。发育中的大鼠脑内PIP和PIP2水平升高，缺血10分钟后降低。