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

Journal of Chromatography. B, Biomedical Applications
D B GoodnoughP L Wood

Abstract

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.

Citations

Jan 16, 1998·European Journal of Pharmacology·J E HawkinsonP L Wood
Jul 1, 1997·Progress in Neurobiology·A Hashimoto, T Oka
Sep 14, 2010·Bioscience, Biotechnology, and Biochemistry·Taketo OmoriNobuyoshi Esaki
Jul 28, 2012·Journal of Chromatography. a·István IliszAntal Péter
Sep 15, 2009·Brain Research·Jordan E AntflickDavid R Hampson
Dec 18, 2001·The Journal of Biological Chemistry·Erica RosemondDavid R Hampson
Jul 1, 2020·Cellular and Molecular Life Sciences : CMLS·Giulia MurtasLoredano Pollegioni
Mar 22, 2014·Analytical Chemistry·David PolcariJanine Mauzeroll
Mar 17, 2009·Biochemical and Biophysical Research Communications·Taketo OmoriNobuyoshi Esaki

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