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Analysis of neuroactive amino acids from microdialysate samples by fluorescence detection using a modification of the 6‐aminoquinolyl‐ N ‐hydroxysuccinimidyl carbamate method
Author(s) -
OreiroGarcía M. Teresa,
VázquezIllanes M. Dolores,
SierraParedes Germán,
SierraMarcuño Germán
Publication year - 2005
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.499
Subject(s) - chemistry , chromatography , derivatization , carbamate , glycine , reproducibility , high performance liquid chromatography , amino acid , fluorescence , serine , biochemistry , physics , quantum mechanics , enzyme
A sensitive and rapid reversed‐phase high‐performance liquid chromatographic method using pre‐column derivatization with 6‐aminoquinolyl‐ N ‐hydroxysuccinimidyl carbamate (AQC) and fluorescence detection is reported. By directly derivatizing microdialysate samples with AQC, an automatic and rapid simultaneous measurement of aspartate, serine, glutamate, glycine and histidine was developed. Excellent linearity ( r 2 ≥ 0.998) was achieved for the standard mixture used for the validation experiments. Within‐day and between‐day precision was less than 6.2%, and the accuracy ranged from 95 to 105.2% in standards. This method is suitable for single run analysis of a high number of small volume microdialysate samples from rat hippocampus. Amino acids from microdialysate samples were quantified with RSD for reproducibility below 2%, and at approximately 0.1% for retention time. Copyright © 2005 John Wiley & Sons, Ltd.