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In vivo simultaneous monitoring of gamma-aminobutyric acid, glutamate, and L-aspartate using brain microdialysis and capillary electrophoresis with laser-induced fluorescence detection: Analytical developments and in vitro/in vivo validations.
Electrophoresis. 2003 Sep; 24(18):3187-96.E

Abstract

gamma-Aminobutyric acid (GABA), glutamate (Glu), and L-aspartate (L-Asp) are three major amino acid neurotransmitters in the central nervous system. In this work, a method for the separation of these three neurotransmitters in brain microdialysis samples using a commercially available capillary electrophoresis (CE) system has been developed. Molecules were tagged on their primary amine function with the fluorogene agent naphthalene-2,3-dicarboxaldehyde (NDA), and, after separation by micellar electrokinetic chromatography, were detected by laser-induced fluorescence using a 442 nm helium-cadmium laser. The separation conditions for the analysis of derivatized neurotransmitters in standard solutions and microdialysates have been optimized, and this method has been validated on both pharmacological and analytical basis. The separation of GABA, Glu, and L-Asp takes less than 10 min by using a 75 mmol/L borate buffer, pH 9.2, containing 70 mmol/L SDS and 10 mmol/L hydroxypropyl-beta-cyclodextrin and + 25 kV voltage. The detection limits were 3, 15 nmol/L and, 5 nmol/L for GABA, Glu, and L-Asp, respectively. Moreover, submicroliter samples can be analyzed. This method allows a simple, rapid and accurate measurement of the three amino acid neurotransmitters for the in vivo brain monitoring using microdialysis sampling.

Authors+Show Affiliations

Laboratoire de Neuropharma- cologie et Neurochimie, INSERM U512, Université Claude Bernard, Lyon, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

14518043

Citation

Sauvinet, Valérie, et al. "In Vivo Simultaneous Monitoring of Gamma-aminobutyric Acid, Glutamate, and L-aspartate Using Brain Microdialysis and Capillary Electrophoresis With Laser-induced Fluorescence Detection: Analytical Developments and in Vitro/in Vivo Validations." Electrophoresis, vol. 24, no. 18, 2003, pp. 3187-96.
Sauvinet V, Parrot S, Benturquia N, et al. In vivo simultaneous monitoring of gamma-aminobutyric acid, glutamate, and L-aspartate using brain microdialysis and capillary electrophoresis with laser-induced fluorescence detection: Analytical developments and in vitro/in vivo validations. Electrophoresis. 2003;24(18):3187-96.
Sauvinet, V., Parrot, S., Benturquia, N., Bravo-Moratón, E., Renaud, B., & Denoroy, L. (2003). In vivo simultaneous monitoring of gamma-aminobutyric acid, glutamate, and L-aspartate using brain microdialysis and capillary electrophoresis with laser-induced fluorescence detection: Analytical developments and in vitro/in vivo validations. Electrophoresis, 24(18), 3187-96.
Sauvinet V, et al. In Vivo Simultaneous Monitoring of Gamma-aminobutyric Acid, Glutamate, and L-aspartate Using Brain Microdialysis and Capillary Electrophoresis With Laser-induced Fluorescence Detection: Analytical Developments and in Vitro/in Vivo Validations. Electrophoresis. 2003;24(18):3187-96. PubMed PMID: 14518043.
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TY - JOUR T1 - In vivo simultaneous monitoring of gamma-aminobutyric acid, glutamate, and L-aspartate using brain microdialysis and capillary electrophoresis with laser-induced fluorescence detection: Analytical developments and in vitro/in vivo validations. AU - Sauvinet,Valérie, AU - Parrot,Sandrine, AU - Benturquia,Nadia, AU - Bravo-Moratón,Eva, AU - Renaud,Bernard, AU - Denoroy,Luc, PY - 2003/10/1/pubmed PY - 2004/6/15/medline PY - 2003/10/1/entrez SP - 3187 EP - 96 JF - Electrophoresis JO - Electrophoresis VL - 24 IS - 18 N2 - gamma-Aminobutyric acid (GABA), glutamate (Glu), and L-aspartate (L-Asp) are three major amino acid neurotransmitters in the central nervous system. In this work, a method for the separation of these three neurotransmitters in brain microdialysis samples using a commercially available capillary electrophoresis (CE) system has been developed. Molecules were tagged on their primary amine function with the fluorogene agent naphthalene-2,3-dicarboxaldehyde (NDA), and, after separation by micellar electrokinetic chromatography, were detected by laser-induced fluorescence using a 442 nm helium-cadmium laser. The separation conditions for the analysis of derivatized neurotransmitters in standard solutions and microdialysates have been optimized, and this method has been validated on both pharmacological and analytical basis. The separation of GABA, Glu, and L-Asp takes less than 10 min by using a 75 mmol/L borate buffer, pH 9.2, containing 70 mmol/L SDS and 10 mmol/L hydroxypropyl-beta-cyclodextrin and + 25 kV voltage. The detection limits were 3, 15 nmol/L and, 5 nmol/L for GABA, Glu, and L-Asp, respectively. Moreover, submicroliter samples can be analyzed. This method allows a simple, rapid and accurate measurement of the three amino acid neurotransmitters for the in vivo brain monitoring using microdialysis sampling. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/14518043/In_vivo_simultaneous_monitoring_of_gamma_aminobutyric_acid_glutamate_and_L_aspartate_using_brain_microdialysis_and_capillary_electrophoresis_with_laser_induced_fluorescence_detection:_Analytical_developments_and_in_vitro/in_vivo_validations_ L2 - https://doi.org/10.1002/elps.200305565 DB - PRIME DP - Unbound Medicine ER -