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MEKC-LIF of gamma-amino butyric acid in microdialysate: systematic optimization of the separation conditions by factorial analysis.
J Neurosci Methods. 2005 Aug 30; 147(1):36-47.JN

Abstract

Micellar electrokinetic chromatography allows the efficient separation of biogenic amines and amino acids in biological samples. Analytes of interest, sample composition, and sample matrix may vary between studies, which necessitates optimization of separations to meet the requirements and conditions of an experiment. Factorial analysis is an efficient tool to accomplish this type of optimization involving multiple interacting factors. The present study describes an optimization procedure for separation of the inhibitory neurotransmitter GABA utilizing capillary electrophoresis with laser induced fluorescence detection. Standards labeled with the flourogenic reagent 3-(2-furoyl)quinoline-2 carboxaldehyde were separated with varying concentrations of borate buffer, beta-cyclodextrin, sodium dodecyl sulfate and pH. The optimized separation method had a correlation coefficient between concentration of GABA and fluorescent signal of 0.98, and was linear in the desired concentration range of 25 nM-10 microM. Glutamic acid, aspartic acid and taurine were also quantified using this separation. When applied to microdialysate collected from the region of the suprachiasmatic nucleus, this separation was able to measure daily variations in GABA levels. The factorial design experiment has proven to be a useful tool, allowing adjustments in the separation of neurotransmitters based on individual requirements.

Authors+Show Affiliations

Department of Biology, Georgia State University, Atlanta, 30302-4098, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

15979724

Citation

Ehlen, J Christopher, et al. "MEKC-LIF of Gamma-amino Butyric Acid in Microdialysate: Systematic Optimization of the Separation Conditions By Factorial Analysis." Journal of Neuroscience Methods, vol. 147, no. 1, 2005, pp. 36-47.
Ehlen JC, Albers HE, Breyer ED. MEKC-LIF of gamma-amino butyric acid in microdialysate: systematic optimization of the separation conditions by factorial analysis. J Neurosci Methods. 2005;147(1):36-47.
Ehlen, J. C., Albers, H. E., & Breyer, E. D. (2005). MEKC-LIF of gamma-amino butyric acid in microdialysate: systematic optimization of the separation conditions by factorial analysis. Journal of Neuroscience Methods, 147(1), 36-47.
Ehlen JC, Albers HE, Breyer ED. MEKC-LIF of Gamma-amino Butyric Acid in Microdialysate: Systematic Optimization of the Separation Conditions By Factorial Analysis. J Neurosci Methods. 2005 Aug 30;147(1):36-47. PubMed PMID: 15979724.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - MEKC-LIF of gamma-amino butyric acid in microdialysate: systematic optimization of the separation conditions by factorial analysis. AU - Ehlen,J Christopher, AU - Albers,H Elliott, AU - Breyer,Emelita D, PY - 2004/10/21/received PY - 2005/03/01/revised PY - 2005/03/03/accepted PY - 2005/6/28/pubmed PY - 2005/10/28/medline PY - 2005/6/28/entrez SP - 36 EP - 47 JF - Journal of neuroscience methods JO - J Neurosci Methods VL - 147 IS - 1 N2 - Micellar electrokinetic chromatography allows the efficient separation of biogenic amines and amino acids in biological samples. Analytes of interest, sample composition, and sample matrix may vary between studies, which necessitates optimization of separations to meet the requirements and conditions of an experiment. Factorial analysis is an efficient tool to accomplish this type of optimization involving multiple interacting factors. The present study describes an optimization procedure for separation of the inhibitory neurotransmitter GABA utilizing capillary electrophoresis with laser induced fluorescence detection. Standards labeled with the flourogenic reagent 3-(2-furoyl)quinoline-2 carboxaldehyde were separated with varying concentrations of borate buffer, beta-cyclodextrin, sodium dodecyl sulfate and pH. The optimized separation method had a correlation coefficient between concentration of GABA and fluorescent signal of 0.98, and was linear in the desired concentration range of 25 nM-10 microM. Glutamic acid, aspartic acid and taurine were also quantified using this separation. When applied to microdialysate collected from the region of the suprachiasmatic nucleus, this separation was able to measure daily variations in GABA levels. The factorial design experiment has proven to be a useful tool, allowing adjustments in the separation of neurotransmitters based on individual requirements. SN - 0165-0270 UR - https://www.unboundmedicine.com/medline/citation/15979724/MEKC_LIF_of_gamma_amino_butyric_acid_in_microdialysate:_systematic_optimization_of_the_separation_conditions_by_factorial_analysis_ DB - PRIME DP - Unbound Medicine ER -