Monitoring of dopamine and its metabolites in brain microdialysates: Method combining freeze-drying with liquid chromatography–tandem mass spectrometry
| Autor: | Věra Bubeníková-Valešová, Marek Kuzma, Petr Kačer, Věra Najmanová, Lukas Rambousek, Romana Šlamberová, Kamila Syslová |
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| Rok vydání: | 2011 |
| Předmět: |
3
4-Dihydroxyphenylacetic acid Dopamine Microdialysis Electrospray ionization Tandem mass spectrometry Sensitivity and Specificity Biochemistry High-performance liquid chromatography Nucleus Accumbens Methamphetamine Analytical Chemistry chemistry.chemical_compound Drug Stability Tandem Mass Spectrometry Liquid chromatography–mass spectrometry Animals 3-Methoxytyramine Solid phase extraction Rats Wistar Chromatography Organic Chemistry Homovanillic acid Reproducibility of Results Homovanillic Acid General Medicine Rats Freeze Drying chemistry Linear Models 3 4-Dihydroxyphenylacetic Acid Female Chromatography Liquid |
| Zdroj: | Journal of Chromatography A. 1218:3382-3391 |
| ISSN: | 0021-9673 |
| DOI: | 10.1016/j.chroma.2011.02.006 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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