Short communication
Optimization of dansyl derivatization and chromatographic conditions in
the determination of neuroactive amino acids of biological samples
Xuejun Kanga,c,*, Jing Xiaoc, Xiao Huangc, Zhongze Gua,b,*
aState Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096, PR China
bCenter of Learning Science, Southeast University, Nanjing, 210096, PR China
cSchool of Public Health, Southeast University, Nanjing, 210009, PR China
Received 7 October 2005; received in revised form 12 November 2005; accepted 14 November 2005
Available online 10 January 2006
Abstract
Background:Amino acid neurotransmitters represent a major class of compounds that are involved in neuronal communication at CNS
synapses.
Methods:Twelve amino acids were separated after precolumn derivatization with dansyl chloride. The biologically important amino acids,
taurine, aspartate, glutamate, glycine, alanine andg-aminobutyric acid were determined in rat brain tissue and rabbit plasma.
Results:The major modifications to previous methods included isocratic elution instead of gradient elution to reduce time and cost of serial
analyses; optimization of mobile phase to improve the separation of free dansyl with derivatives, so as to elide complex steps for the clean of
the chromatogram; and selection of an internal standard (Trazodone) to improve the reproducibility and the reliability of procedures. Twelve
amino acids were assayed within 35 min. Othera-amino acids that are relevant to the make-up of mammalian proteins did not interfere with
the determination. The standard curve, recovery, analytical precision and detection limits for each neuroactive amino acid were determined.
Conclusion:This assay separated 12 amino acids in a single run. Six neuroactive amino acids were also simultaneous measured by isocratic
HPLC with UV detection. The method is applicable in determination of Tau, Glu, Asp, Gly, Ala and GABA in biological samples.

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