Determination of reduced-form glutathione and total glutathione in blood and plasma by high performance liquid chromatography with on-column fluorescence derivatization

被引：9

作者：

Fukunaga, K ^{[1
]}

Nakazono, N

Yoshida, M

机构：

[1] Kansai Med Univ, Dept Publ Hlth, Osaka 5708506, Japan

[2] Kansai Univ, Fac Engn, Dept Biotechnol, Osaka 5648680, Japan

来源：

CHROMATOGRAPHIA | 1998年 / 48卷 / 9-10期

关键词：

column liquid chromatography; on-column fluorescence derivatization; blood and plasma; glutathione; ortho-phthalaldehyde;

D O I：

10.1007/BF02467600

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

An on-column fluorometric derivatization method has been developed for the determination of reduced-form glutathione and total glutathione in blood and plasma by high-performance liquid chromatography. Blood hemolysate or diluted plasma was deproteinized with perchloric acid. For total glutathione determination, any oxidized forms of glutathione in samples were converted to reduced-form glutathione with dithiothreitol prior to protein precipitation. Deproteinized samples were injected without pretreatment. Glutathione was fluorescence-derivatized and separated on a C18-bonded vinyl alcohol polymer column with ortho-phthalaldehyde in sodium berate buffer-acetonitrile, as mobile phase: the ortho-pathalaldehyde-glutathione, fluorescent isoindole adduct is measured by its fluorescence response. Due to the high sensitivity, selectivity reproducibility and the simple HPLC system, the method is particularly suitable for routine assay of numerous or small amounts of samples.

引用

页码：690 / 694

页数：5

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