摘要
建立一种高灵敏度的氯甲酸芴甲酯柱前衍生荧光检测反相高效液相色谱法测定血清中γ氨基丁酸的方法．内标为己氨酸，固定相为ＳｈｉｍＰａｃｋＣＬＣＯＤＳ（Ｍ），４６ｍｍ×１５０ｍｍ，填充料为５μｍ；流动相采用二元梯度洗脱，Ａ相为００５ｍｏｌ／Ｌ醋酸钠缓冲液∶水∶四氢呋喃∶冰乙酸（２５０∶１００∶１５∶２２），Ｂ相为乙腈∶甲醇（４∶１）．系统研究了ｐＨ值、反应时间、离子强度及衍生化试剂用量对衍生化反应的影响，确定最佳反应条件和最佳色谱条件．方法的精密度为批内变异系数＜４６％，批间变异系数＜６１％；最低检出限（信噪比＝２）为３１ｎｍｏｌ／Ｌ；线性范围为５０～１０００ｎｍｏｌ／Ｌ，γ２＝０９９９２；平均回收率为９７１％．
A new highly sensitive HPLC method for determination γ aminobutyric acid(GABA) in serum was developed by precolumn derivatization of GABA with 9 fluorenylmethyl chlorormate(FMOC Cl) using ε amino n caproic acid as an internal standard. The column used was Shim Pack CLC ODS(M), 4 6 mm×150 mm,5 μm. Mobile phase A consisted of sodium acetate buffer (0 05 mol/L,pH 3 65)/water/tetranhydrofunan/glacial acetic acid(250/100/15/2 2).Mobile phase B was acetonitrile/methanol (4/1). The conditions of derivatization and chromatography were studied to find a optimal procedure,which included pH value, reaction time, ionic strength and amount of derivatizing reagent.The coefficient of variance of the method was less than 4 6% for within runs, 6 1% for between runs, respectively. The minimal detection limit ( S/N =2) was 3 1 nmol/L . The linearity was observed from 10 to 1 000 nmol/L, and the coefficient of determination was 0 9992. The average recovery was 97 1%.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1998年第6期543-547,共5页
Progress In Biochemistry and Biophysics
基金
卫生部科研基金
关键词
Γ-氨基丁酸
反相高效液相色谱法
氯甲酸芴甲酯
γ aminobutyric acid(GABA), reversed phase high performance liquid chromatography,9 fluorenylmethyl chlorormate(FMOC Cl)
