高效液相色谱-荧光法测定大鼠纹状体细胞外液中4种氨基酸的含量  

HPLC-RF determination of four amino acids levels in striatal extracellular fluid

作　　者：刘春丽[1] 陈虹[1] 钟明[1] 马静怡[1] 丁慧[1] 张万鑫[1] 杨赞章[1] 

机构地区：[1]石河子大学药学院教育部新疆特种植物药资源重点实验室,石河子832002

出　　处：《药物分析杂志》2012年第11期1914-1918,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金资助项目(NO:30860334;30560171);石河子大学自然科学与技术创新重点项目(NO:ZRKX2009ZD05)

摘　　要：目的:采用高效液相色谱-荧光检测法(HPLC-RF)建立大鼠脑微透析液中天门冬氨酸(Asp)、谷氨酸(Glu)、甘氨酸(Gly)和γ-氨基丁酸(GABA)的测定方法。方法:采用ODS-C18(5μm,250 mm×4.6 mm)色谱柱,柱温为35℃,流动相A为磷酸盐缓冲液(pH 6.7),流动相B为纯甲醇,梯度洗脱,流速为1.0 mL.min-1,荧光检测器的激发波长为239 nm,发射波长为375 nm。以邻苯二甲醛和β-巯基乙醇为衍生化试剂,柱前衍生,将脑微透析液直接注入高效液相色谱仪完成分析。结果:Asp、Glu、Gly和GABA 4种氨基酸在0.5～32μmol.L-1范围内线性关系良好,回收率范围为96.2%～102.1%。结论:所建立的氨基酸检测方法具有流动相配制简单、保留时间稳定、分析时间短、重复性好和回收率高等优点,可用于脑微透析液中痕量氨基酸的测定。Objective ：To develop an HPLC - RF method for Asp, Glu, Gly and GABA determination in rat cerebral microdialysates. Methods：ODS- Cls （5 μm,250 mm × 4. 6 mm） column was employed by a gradient elution, u- sing phosphate buffer solution （ pH 6. 7 ） as mobile phase A and pure methanol as mobile phase B at the column temperature of 35 ℃ and the flow rate of 1 mL min - 1. Linear gradient elution was adopted with a florescent detector （ λex = 239 nm, λcm = 375 nm）. O - phthaldialdehyde/β - mercaptoethanol was used for the derivatization of ami- no acids. Then HPLC - RF was used to measure the striatal extracellular levels of four amino acids. Results： These four components were all found to have good linear relationship over a concentration range of 0. 5 -32 μmol L- 1, and the recoveries of four amino acids were in the range of 96.2% - 102. 1%. Conclusions： The HPLC - RF meth- od has the advantages of simple preparation, constant retention time, short analysis time, good repeatability and high recovery rate, which can be used for cerebral microdialysate trace determination of amino acids.

关 键 词：脑微透析 纹状体 高效液相色谱-荧光检测法 氨基酸类递质 衍生化 邻苯二甲醛(OPA) 天门冬氨酸 谷氨酸 甘氨酸 氨基丁酸 

分 类 号：R917[医药卫生—药物分析学]