Simultaneous determination of D- and L-serine in rat brain microdialysis sample using a column-switching HPLC with fluorimetric detection

Biomed Chromatogr. 2004 Dec;18(10):813-9. doi: 10.1002/bmc.394.


Both D- and L-serine in rat brain microdialysis sample were simultaneously determined by pre-column fluorescence derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), separation of the derivatives on ODS column, TSKgel ODS-80TsQA, followed by Pirkle type chiral columns, Sumichiral OA-2500 (S), which gave a sufficient enantiomeric separation of NBD-D-serine and NBD-L-serine, and fluorimetric detection at a wavelength of 540 nm with an excitation wavelength of 470 nm. The peaks of NBD-D-serine and NBD-L-serine in the rat brain microdialysis sample were clearly found, and the validation study showed satisfactory results; the precision and accuracy were within 5.14 and 109%, respectively. Using the proposed HPLC method, the time-course profile of D-serine concentration in rat prefrontal cortex following intraperitoneal administration of D-serine was investigated. As a consequence, D-serine appeared to be rapidly distributed in the brain, and then decreased gradually with time in the extracellular fluid of the rat prefrontal cortex. The proposed HPLC method will be useful for in vivo studies on D-serine, which acts as a coagonist for N-methyl-D-aspartate receptor, to the extracellular fluid of rat brain.

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan / analogs & derivatives*
  • 4-Chloro-7-nitrobenzofurazan / chemistry
  • Animals
  • Brain Chemistry*
  • Chromatography, High Pressure Liquid / methods*
  • Fluorometry
  • Kinetics
  • Male
  • Microdialysis*
  • Prefrontal Cortex / chemistry
  • Prefrontal Cortex / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Serine / analysis*
  • Serine / chemistry*
  • Serine / pharmacokinetics
  • Spectrometry, Fluorescence
  • Stereoisomerism


  • 7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole
  • Serine
  • 4-Chloro-7-nitrobenzofurazan