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Simultaneous determination of D ‐ and L ‐serine in rat brain microdialysis sample using a column‐switching HPLC with fluorimetric detection
Author(s) -
Fukushima Takeshi,
Kawai Junko,
Imai Kazuhiro,
Toyo'oka Toshimasa
Publication year - 2004
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.394
Subject(s) - microdialysis , chemistry , serine , chromatography , derivatization , high performance liquid chromatography , enantiomer , extracellular , biochemistry , stereochemistry , phosphorylation
Both d ‐ and l ‐serine in rat brain microdialysis sample were simultaneously determined by pre‐column uorescence derivatization with 4‐uoro‐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 sufcient enantiomeric separation of NBD– d ‐serine and NBD‐ l ‐serine, and uorimetric 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 prole 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 uid 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 uid of rat brain. Copyright © 2004 John Wiley & Sons, Ltd.