豚鼠脑在体微透析液氨基酸AccQ·Tag HPLC荧光测定法

目的建立柱前6-氨基喹啉基-N-羟基琥珀酰亚胺基氨基甲酸酯(AQC)衍生高效液相色谱(HPLC)脑在体微透析液中痕量氨基酸的检测方法。方法采用AQC为柱前衍生剂,氨基酸专用分析柱,二元梯度洗脱,荧光检测方法分析样品中的氨基酸。结果该法对脑微透析液中17种氨基酸分离较好,相关系数为0.902 5~0.999 5,平均回收率为94.24%~98.34%,平均CV小于5%。在50 m in内可完成微透析液中17种氨基酸的测定。结论该方法稳定性好、敏感性和准确性较高,适用于豚鼠脑在体微透析液中痕量氨基酸水平的测定。

Determination of the Concentration of Amino-acid in Vivo Microdialysis Fluid of Guinea Pigs Brain by AccQ·Tag HPLC Fluorescence Analysis Method

Objective To establish a method for determination of amino acid in vivo microdialysis fluid of guinea pigs brain by HPLC with per column derivatization.Methods The microdialysis fluidswere derived with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate(AQC) reagent and the derivatives were separated by a two gradient elution and detected byfluorescence detector.Results The 17 amino acid were separated better.The correlation coefficients of the calibration graph for 16 aminoacid were good,the recoveries range were between 94.24% and 98.34%,the CV of all amino acid were less than 5%.Conclusion The methodis used to determine the amino acid in microdialysis fluid of brain,which showedthat it is a simple,convenient and sensitive method.