Determination of free-form amphetamine in rat brain by ion-pair liquid chromatography-electrospray mass spectrometry with in vivo microdialysis
| Autor: | Chiuan-Hung Haung, Shiang-Ling Lin, Ming-Ren Fuh, Wynn H.T. Pan |
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| Rok vydání: | 2004 |
| Předmět: |
Male
Electrospray Microdialysis Spectrometry Mass Electrospray Ionization Mass spectrometry Biochemistry High-performance liquid chromatography Analytical Chemistry Rats Sprague-Dawley chemistry.chemical_compound Trifluoroacetic acid Animals Detection limit Chromatography Chemistry Organic Chemistry Brain Reproducibility of Results General Medicine Reversed-phase chromatography Reference Standards Chromatography Ion Exchange Rats Solutions Amphetamine Central Nervous System Stimulants Quantitative analysis (chemistry) |
| Zdroj: | Journal of chromatography. A. 1031(1-2) |
| ISSN: | 0021-9673 |
| Popis: | An ion-pair liquid chromatography-electrospray mass spectrometry (LC-ESI-MS) method with in vivo microdialysis for the determination of free-form amphetamine in rat brain has been developed. A microdialysis probe was surgically implanted into the striatum of the rat and artificial cerebrospinal fluid (aCSF) was used as the perfusion medium. Samples were collected and then analyzed off-line by LC-ESI-MS. A reversed phase C18 column was employed for LC separation. Trifluoroacetic acid (TFA) was added in the mobile phase (acetonitrile-water, 10:90, v/v) as an ion-pair reagent. The ion-pair process disguises the protonated amphetamine cations from the ESI-MS electric field as neutral molecules. Post-column addition of volatile organic acid was utilized to minimize TFA signal suppression effect on ESI-MS detection. More than six-fold enhancement of ESI-MS response was achieved by the post-column addition of propionic acid. Good linearity (0.01-1.00 microg/ml, r2 = 0.99) and detection limit (0.002 microg/ml) were determined. Good precision and accuracy were obtained. The applicability of this newly developed method was demonstrated by continuous monitoring of amphetamine concentrations in rat brain after a single 3.0 mg/kg i.p. administration. |
| Databáze: | OpenAIRE |
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