A high-throughput on-line microdialysis-capillary assay for D-serine.
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.
O'Brien
Kylie B
KB
Department of Chemistry, University of Minnesota, Minneapolis 55455, USA.
Esguerra
Manuel
M
Klug
Christina T
CT
Miller
Robert F
RF
Bowser
Michael T
MT
D000818:Animals; D019075:Electrophoresis, Capillary / Q000379:methods*; D017551:Microdialysis; D018377:Neurotransmitter Agents / Q000032:analysis; D009862:Online Systems; D012160:Retina / Q000737:chemistry; D012694:Serine / Q000032:analysis*; D013237:Stereoisomerism; D013997:Time Factors; D014562:Caudata
D018377:Neurotransmitter Agents; D012694:Serine