Establishment of HPLC technology of precolumn derivatization with PITC in determining contents of Glu, Gly and GABA in brain tissues of rats with epilepsy
LI Min
PAN Qun-wan
WANG Hai-hua
WANG Lie-cheng
Abstract：Objective To establish high-performance liquid chromatography ( HPLC) technology of precolumn derivat-ization with phenyl isothiocyanate ( PITC) to determine the contents of glutamic acid ( Glu) , glycine ( Gly) andγ-aminobu-tyric acid ( GABA) in the brain tissues of rats with epilepsy, and to verify the results.Methods Kainic acid ( KA) was in-jected into the hippocampus CA3 region of rats to make the epilepsy models.Two weeks later, we obtained the hippocampus and cortex tissues, respectively.The diluted samples were precolumn derivatizated with PITC.The mobile phase consisted of sodium acetate and acetonitrile gradient elution.We calculated the contents of Glu, Gly and GABA in hippocampus and cor-tex by venusil-AA column combined with UV detector.Results The amino acids were separated within 30 min.A good line-arity was obtained from (0.20-12.50) mg/L of Glu, the correlation coefficient was 0.999 3, the intraday RSD was 1.46%, interday RSD was 1.84%, and the average recovery rate was 97.12%±2.98%.A good linearity was obtained from (0.01-6.25) mg/L of Gly, the correlation coefficient was 0.998 4, the intraday RSD was 1.17%, interday RSD was 1.62%, and the average recovery rate was 96.63%±1.28%.A good linearity was obtained from (0.10-6.25) mg/L of GABA, the cor-relation coefficient was 0.999 4, the intraday RSD was 1.00%, interday RSD was 1.53%, and the average recovery rate was 99.25%±2.98%.Conclusion The HPLC technology of precolumn derivatization with PITC in determining the contents of Glu, Gly and GABA in the brain tissues of rats with epilepsy is successfully established.
Keywords：epilepsyhigh-performance liquid chromatographyphenyl isothiocyanateprecolumn derivatizationglu-tamic acidglycineγ-aminobutyric acidrats
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 8-11 )