Analysis of Intact Glucuronides and Sulfates of Serotonin, Dopamine, and Their Phase I Metabolites in Rat Brain Microdialysates by Liquid Chromatography-Tandem Mass Spectrometry

被引：66

作者：

Uutela, Paivi ^{[1
]}

Reinila, Ruut ^{[1
]}

Harju, Kirsi ^{[1
]}

Piepponen, Petteri ^{[2
]}

Ketola, Raimo A. ^{[3
]}

Kostiainen, Risto ^{[1
]}

机构：

[1] Univ Helsinki, Fac Pharm, Div Pharmaceut Chem, FI-00014 Helsinki, Finland

[2] Univ Helsinki, Fac Pharm, Div Pharmacol & Toxicol, FI-00014 Helsinki, Finland

[3] Univ Helsinki, Fac Pharm, CDR, FI-00014 Helsinki, Finland

来源：

ANALYTICAL CHEMISTRY | 2009年 / 81卷 / 20期

基金：

芬兰科学院;

关键词：

UDP-GLUCURONOSYLTRANSFERASES; DIHYDROXYPHENYLACETIC ACID; CEREBROSPINAL-FLUID; HUMAN LIVER; 5-HYDROXYTRYPTAMINE; CATECHOLAMINES; IDENTIFICATION; SUPERFAMILY; CONJUGATION; EXPRESSION;

D O I：

10.1021/ac901320z

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A method for the analysis of intact glucuronides and sulfates of common neurotransmitters serotonin (5-HT) and dopamine (DA) as well as of 5-hydroxy-3-indoleacetic acid (5-HIAA), 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA) in rat brain microdialysates by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. Enzyme-assisted synthesis using rat liver microsomes as a biocatalyst was employed for the production of 5-HT-, 5-HIAA-, DOPAC-, and HVA-glucuronides for reference compounds. The sulfate conjugates were synthesized either chemically or enzymatically using a rat liver S9 fraction. The LC-MS/MS method was validated by determining the limit,; of detection and quantitation, linearity, and repeatability for the quantitative analysis of 5-HT and DA and their glucuronides, as well as of 5-HLAA, DOPAC, and HVA and their sulfate-conjugates. In this study, 5-HT-glucuronide was for the first time detected in rat brain. The concentration of 5-HT-glucuronide (1.0-1.7 nM) was up to 2.5 times higher than that of free 5-HT (0.4-2.1 nM) in rat brain microdialysates, whereas the concentration of DA-glucuronide (1.0-1.4 nM) was at the same level or lower than the free DA (1.2-2.4 nM). The acidic metabolites of neurotransmitters, 5-HLAA, HVA, and DOPAC, were found in free and sulfated form, whereas their glucuronidation was not observed.

引用

页码：8417 / 8425

页数：9

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