Simultaneous determination of D- and L-serine in rat brain microdialysis sample using a column-switching HPLC with fluorimetric detection
Simultaneous determination of D- and L-serine in rat brain microdialysis sample using a column-switching HPLC with fluorimetric detection
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DOI：
10.1002/bmc.394
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发表时间：
2004-12-01
影响因子：
1.8
通讯作者：
Toyo'oka, T
中科院分区：
文献类型：
--
作者：
Fukushima, T;Kawai, J;Toyo'oka, T
采用4-氟-7-硝基-2,1,3-苯并二唑（NBD-F）柱前荧光衍生法同时测定大鼠脑微透析样品中的D和l -丝氨酸，衍生物在ODS色谱柱TSKgel ODS80TsQA上分离，再用Pirkle型手性色谱柱Sumichiral -2500 (S)进行分离，对nbd -D-丝氨酸和nbd - l -丝氨酸进行充分的对映体分离，荧光检测波长为540 nm，激发波长为470 nm。大鼠脑微透析样品中nbd - d -丝氨酸和nbd - l -丝氨酸峰清晰可见，验证研究结果令人满意；精密度和准确度分别在5.14%和109%以内。采用高效液相色谱法，研究了d -丝氨酸腹腔注射后大鼠前额皮质d -丝氨酸浓度的时间谱。结果表明，d -丝氨酸在大鼠前额叶皮层细胞外液中呈现快速分布，然后随时间逐渐减少的趋势。该方法可用于大鼠脑细胞外液中n -甲基- d -天冬氨酸受体凝聚剂d -丝氨酸的体内研究。版权所有：John Wiley Sons， Ltd. 2004
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 ODS80TsQA, 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. Copyright (C) 2004 John Wiley Sons, Ltd.