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In vivo monitoring of amino acids by microdialysis sampling with on-line derivatization by naphthalene-2,3-dicarboxyaldehyde and rapid micellar electrokinetic capillary chromatography.
J Neurosci Methods. 2004 Sep 30; 138(1-2):189-97.JN

Abstract

An analytical method was developed to monitor amino acids collected by in vivo microdialysis. Microdialysate was continuously derivatized on-line by mixing 6 mM naphthalene-2,3-dicarboxyaldehyde (NDA) and 10 mM potassium cyanide with the dialysate stream in a fused silica capillary to form fluorescent products. Reaction time, determined by the flow rate and volume of reaction capillary, was 3 min. Derivatized amino acids were continuously delivered into a flow-gated interface and periodically injected onto a capillary electrophoresis unit equipped with a laser-induced fluorescence detection based on a commercial microscope. Separation was performed in the micellar electrokinetic chromatography mode using 30 mM sodium dodecyl sulfate in 15 mM phosphate buffer at pH 8.0 as the separation media. An electric field of 1.3 kV/cm was applied across a 10 cm long, 10 microm internal diameter separation capillary. These conditions allowed 17 amino acid derivatives to be resolved in less than 30 s. On-line injections could be performed at 30 s intervals for in vivo samples. Detection limits were from 10 to 30 nM for the amino acids. The method was applied to monitor the acute ethanol-induced amino acid level changes in freely moving rats. The results demonstrate the utility of the method to reveal dynamics of amino acid concentration in vivo.

Authors+Show Affiliations

Shou M
Department of Chemistry, University of Michigan, 930 North University Avenue, Ann Arbor, MI 48109, USA.
Smith AD
No affiliation info available
Shackman JG
No affiliation info available
Peris J
No affiliation info available
Kennedy RT
No affiliation info available

MeSH

Amino AcidsAnimalsBiosensing TechniquesBrain ChemistryChromatography, Micellar Electrokinetic CapillaryDose-Response Relationship, DrugLasersMaleMicrodialysisNaphthalenesNeurotransmitter AgentsOnline SystemsPotassium CyanideRatsRats, Sprague-DawleyReaction TimeSodium Dodecyl SulfateSpectrometry, FluorescenceTime Factors

Pub Type(s)

Comparative Study
Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

15325127

Citation

Shou, Minshan, et al. "In Vivo Monitoring of Amino Acids By Microdialysis Sampling With On-line Derivatization By Naphthalene-2,3-dicarboxyaldehyde and Rapid Micellar Electrokinetic Capillary Chromatography." Journal of Neuroscience Methods, vol. 138, no. 1-2, 2004, pp. 189-97.
Shou M, Smith AD, Shackman JG, et al. In vivo monitoring of amino acids by microdialysis sampling with on-line derivatization by naphthalene-2,3-dicarboxyaldehyde and rapid micellar electrokinetic capillary chromatography. J Neurosci Methods. 2004;138(1-2):189-97.
Shou, M., Smith, A. D., Shackman, J. G., Peris, J., & Kennedy, R. T. (2004). In vivo monitoring of amino acids by microdialysis sampling with on-line derivatization by naphthalene-2,3-dicarboxyaldehyde and rapid micellar electrokinetic capillary chromatography. Journal of Neuroscience Methods, 138(1-2), 189-97.
Shou M, et al. In Vivo Monitoring of Amino Acids By Microdialysis Sampling With On-line Derivatization By Naphthalene-2,3-dicarboxyaldehyde and Rapid Micellar Electrokinetic Capillary Chromatography. J Neurosci Methods. 2004 Sep 30;138(1-2):189-97. PubMed PMID: 15325127.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - In vivo monitoring of amino acids by microdialysis sampling with on-line derivatization by naphthalene-2,3-dicarboxyaldehyde and rapid micellar electrokinetic capillary chromatography. AU - Shou,Minshan, AU - Smith,Anthony D, AU - Shackman,Jonathan G, AU - Peris,Joanna, AU - Kennedy,Robert T, PY - 2003/12/19/received PY - 2004/03/16/revised PY - 2004/04/01/accepted PY - 2004/8/25/pubmed PY - 2004/11/16/medline PY - 2004/8/25/entrez SP - 189 EP - 97 JF - Journal of neuroscience methods JO - J Neurosci Methods VL - 138 IS - 1-2 N2 - An analytical method was developed to monitor amino acids collected by in vivo microdialysis. Microdialysate was continuously derivatized on-line by mixing 6 mM naphthalene-2,3-dicarboxyaldehyde (NDA) and 10 mM potassium cyanide with the dialysate stream in a fused silica capillary to form fluorescent products. Reaction time, determined by the flow rate and volume of reaction capillary, was 3 min. Derivatized amino acids were continuously delivered into a flow-gated interface and periodically injected onto a capillary electrophoresis unit equipped with a laser-induced fluorescence detection based on a commercial microscope. Separation was performed in the micellar electrokinetic chromatography mode using 30 mM sodium dodecyl sulfate in 15 mM phosphate buffer at pH 8.0 as the separation media. An electric field of 1.3 kV/cm was applied across a 10 cm long, 10 microm internal diameter separation capillary. These conditions allowed 17 amino acid derivatives to be resolved in less than 30 s. On-line injections could be performed at 30 s intervals for in vivo samples. Detection limits were from 10 to 30 nM for the amino acids. The method was applied to monitor the acute ethanol-induced amino acid level changes in freely moving rats. The results demonstrate the utility of the method to reveal dynamics of amino acid concentration in vivo. SN - 0165-0270 UR - https://www.unboundmedicine.com/medline/citation/15325127/In_vivo_monitoring_of_amino_acids_by_microdialysis_sampling_with_on_line_derivatization_by_naphthalene_23_dicarboxyaldehyde_and_rapid_micellar_electrokinetic_capillary_chromatography_ DB - PRIME DP - Unbound Medicine ER -
Grapherence [↓13]

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