豚鼠脑在体微透析液氨基酸AccQ·Tag HPLC荧光测定法

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

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。结果谊法对脑微透析液中17种氨基酸分离较好，相关系数为0．9025～0．9995，平均回收率为94．24％～98．34％，平均CV小于5％。在50min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。

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 fluids were derived with 6-aminoquinolyl-N-hydroxysuccinimidyl earbamate （AQC） reagent and the derivatives were separated by a two gradient elution and detected by fluorescence detector. Results The 17 amino acid were separated better. The correlation coefficients of the calibration graph for 16 amino acid were good,the recoveries range were between 94.24% and 98.34% ,the CV of all amino acid were less than 5%. Conclusion The method is used to determine the amino acid in microdialysis fluid of brain,which showed that it is a simple,convenient and sensitive method.