A high-throughput on-line microdialysis-capillary assay for D-serine

被引:55
作者
O'Brien, KB
Esguerra, M
Klug, CT
Miller, RF
Bowser, MT
机构
[1] Univ Minnesota, Dept Chem, Minneapolis, MN 55455 USA
[2] Univ Minnesota, Dept Neurosci, Minneapolis, MN 55455 USA
关键词
amino acid detection; enantiomer separation; neurotransmitter; retina;
D O I
10.1002/elps.200390158
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A high-throughput method is described for the analysis Of D-serine and other neurotransmitters in tissue homogenates. Analysis is performed by microdialysis-capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection in a sheath flow detection cell. Sample pretreatment is not required as microdialysis sampling excludes proteins and cell fragments. Primary amines are derivatized on-line with o-phthaldialdehyde (OPA) in the presence of beta-mercaptoethanol followed by on-line CE-LIF analysis. Under the separation conditions described here, D-serine is resolved from L-serine and other primary amines commonly found in biological samples. Each separation requires less than 22 s. Eliminating the need for sample pretreatment and performing the high-speed CE analysis on-line significantly reduces the time required for D-serine analysis when compared with traditional methods. This method has been used to quantify D-serine levels in larval tiger salamander retinal homogenates, as well as dopamine, gamma-amino-n-butyric acid (GABA), glutamate and L-aspartate. D-serine release from an intact retina was also detected.
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页码:1227 / 1235
页数:9
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