DNFB柱前衍生化RP-HPLC测定大鼠肝组织18种氨基酸  

Determination of 18 Amino Acids in Rat Liver with DNFB Pre-column Derivatization by Reversed-Phase High Performance Liquid Chromatography

作　　者：王小琴[1] 邹玉安[2] 郭春燕[1] 薛茜[2] 

机构地区：[1]河北北方学院研究生部,河北张家口075000 [2]河北北方学院附属第一医院,河北张家口075000

出　　处：《河北北方学院学报（自然科学版）》2015年第5期4-8,共5页Journal of Hebei North University:Natural Science Edition

基　　金：河北省卫生厅资助项目(20090591)

摘　　要：目的采用柱前衍生RP-HPLC法测定大鼠肝脏组织中18种氨基酸的含量。方法以2,4-二硝基氟苯(DNFB)为衍生化试剂,采用Hypersil BDS C18(4.6mm×250mm,5μm)色谱柱,以甲醇作为流动相A,0.015mol·L-1醋酸钠缓冲液(pH=6.0)作为流动相B,梯度洗脱,柱温30℃,流速为1ml·min-1,检测波长360nm。结果在30min内大鼠肝脏组织中的18种氨基酸可以得到完全分离,且各氨基酸浓度与峰面积之间具有良好的线性关系,回收率在88.3%~110%之间。结论该方法简便、快捷、灵敏、准确,有高度的稳定性和很好的重复性。可以为其他生物样品氨基酸测定提供参考。Objective To establish a method for the determination of 18 amino acids in rat hepatic tis-sue by RP-HPLC.Methods The samples were pre-column derivated with DNFB,analyzed by a Hypersil BDS C18 column (4.6 mm×200 mm,5 μm)as stationary phase,methanol as mobile phase A and 0.015 mol/L acetate buffer solution (pH=6.0)as mobile phase B in the gradient mode at a flow rate of 1 ml/min.The column temperature was 30 ℃,and the detection wavelength was set at 360 nm.Results The results showed that 18 amino acids in rat hepatic tissue were separated within 30 minutes with a good line-arity.The average recoveries were 88.3%~110%.Conclusion This method is convenient,sensitive,ac-curate,highly steady and of good reproducibility.It may also serve as a good reference for the determina-tion of amino acids in other materials.

关 键 词：肝组织 氨基酸 2 4-二硝基氟苯 RP-HPLC 

分 类 号：R285.5[医药卫生—中药学] O657.72[医药卫生—中医学]