Separation and quantification of D- and L-phosphoserine in rat brain using N alpha-(2,4-dinitro-5-fluorophenyl)-L-alaninamide (Marfey's reagent) by high-performance liquid chromatography with ultraviolet detection.
D-Serine has recently been described to be present in the brain at high concentrations. However, while prior research has demonstrated that L-phosphoserine is the major precursor of L-serine in the brain, the possible role of D-phosphoserine as the direct precursor of D-serine is unknown. To address this problem, we developed an assay to separate and quantitate D- and L-phosphoserine. A very simple HPLC-UV procedure for the separation and quantification of D- and L-phosphoserine is presented using precolumn derivatization with a chiral reagent, N alpha-(2,4-dinitro-5-fluorophenyl)-L-alaninamide (Marfey's reagent), and a conventional C18 reversed-phase column. The procedure is sensitive to 11 pmol on-column and derivatives are stable for at least two weeks at room temperature. Rat brain regions (cortex, hippocampus, striatum, and cerebellum) were analyzed for the presence of D- and L-phosphoserine. It was determined that the brain regions studied contained exclusively L-phosphoserine.
Goodnough
D B
DB
CoCensys Inc., Irvine, calillllll
Lutz
M P
MP
Wood
P L
PL
D000409:Alanine / Q000031:analogs & derivatives*; D000818:Animals; D001923:Brain Chemistry; D002531:Cerebellum / Q000737:chemistry; D002540:Cerebral Cortex / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D003342:Corpus Striatum / Q000737:chemistry; D004136:Dinitrobenzenes; D006624:Hippocampus / Q000737:chemistry; D007202:Indicators and Reagents; D008297:Male; D010768:Phosphoserine / Q000032:analysis* / Q000302:isolation & purification; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D013237:Stereoisomerism
D004136:Dinitrobenzenes; D007202:Indicators and Reagents; D010768:Phosphoserine; C047134:Marfey's reagent; D000409:Alanine