Monitoring of dopamine and its metabolites in brain microdialysates: method combining freeze-drying with liquid chromatography-tandem mass spectrometry
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
21397906
DOI
10.1016/j.chroma.2011.02.006
PII: S0021-9673(11)00188-9
Knihovny.cz E-resources
- MeSH
- Chromatography, Liquid methods MeSH
- Dopamine analogs & derivatives analysis metabolism MeSH
- Rats MeSH
- 3,4-Dihydroxyphenylacetic Acid analysis metabolism MeSH
- Homovanillic Acid analysis metabolism MeSH
- Linear Models MeSH
- Freeze Drying MeSH
- Methamphetamine administration & dosage MeSH
- Microdialysis MeSH
- Nucleus Accumbens chemistry metabolism MeSH
- Rats, Wistar MeSH
- Reproducibility of Results MeSH
- Sensitivity and Specificity MeSH
- Drug Stability MeSH
- Tandem Mass Spectrometry methods MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Dopamine MeSH
- 3,4-Dihydroxyphenylacetic Acid MeSH
- Homovanillic Acid MeSH
- Methamphetamine MeSH
A sensitive assay method was developed for a parallel, rapid and precise determination of dopamine and its metabolites, homovanillic acid, 3-methoxytyramine and 3,4-dihydroxyphenylacetic acid, from brain microdialysates. The method consisted of a pre-treatment step, freeze-drying (lyophilization), to concentrate dopamine and its metabolites from the microdialysates, and a detection step using liquid chromatography combined with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). In particular, the reaction monitoring mode was selected for its extremely high degree of selectivity and the stable-isotope-dilution assay for its high precision of quantification. The developed method was characterized by the following parameters: the precision of the developed method was determined as ≥88.6% for dopamine, ≥89.9% for homovanillic acid, ≥86.1% for 3-methoxytyramine and ≥88.1% for 3,4-dihydroxyphenylacetic acid; the mean accuracy was determined as ≥88.2% for dopamine, ≥88.3% for homovanillic acid, ≥85.9% for 3-methoxytyramine and ≥88.6% for 3,4-dihydroxyphenylacetic acid. The developed method was compared to (1) other combinations of pre-treatment methods (solid phase extraction and nitrogen stripping) with LC-MS and (2) another detection method, liquid chromatography, with electrochemical detection. The novel developed method using combination of lyophilization with LC-ESI-MS/MS was tested on real samples obtained from the nucleus accumbens of rat pups after an acute methamphetamine administration. It was proven that the developed assay could be applied to both a simultaneous analysis of all four substrates (dopamine, homovanillic acid, 3-methoxytyramine and 3,4-dihydroxyphenylacetic acid) in microdialysis samples acquired from the rat brain and the monitoring of their slight concentration changes on a picogram level over time following methamphetamine stimulus.
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