Determination of 23 Amino Acids and Amines in Plasma and Cortex of C57BL/6 Mice with HPLC-FLD.

Abstract

Using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate as a pre-column derivatization reagent, optimized derivatization and chromatography parameters, a simple high-performance liquid chromatography fluorescence detector (HPLC-FLD) method was developed and validated to determine 23 related amines in plasma and cortex of C57BL/6 mice with cerebral ischemia-reperfusion injury. The prepared samples were separated on a ZORBAX SB-C18 column (4.6mm × 250mm, 5μm) with 60% acetonitrile (ACN) and 20mM sodium acetate solution (pH adjusted to 5.0 by phosphoric acid). All analytes achieved good separation within 1.2h at a flow rate of 1mL/min. The limits of detection and limits of detection quantitation of the method were ranged from (0.1-9.2) to (0.3-30.6) ng/mL, respectively. The analytical method was apt for simultaneously determining 23 amino acids in plasma and cortex. Our results revealed that the relevant amino acids were significantly altered (P < 0.05) in C57BL/6 mice.