2D-HPLC同时测定小鼠胸腺中的酸性氨基酸异构体
Simultaneous determination of acidic amino acid enantiomers in the mice thymus by 2D-HPLC
作者:韩海;王启钦;王欢;
Author:
收稿日期: 年卷(期)页码:2015,30(01):-84-86
期刊名称:华西药学杂志
Journal Name:WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字:酸性氨基酸;异构体;离线二维高效液相色谱法;胸腺;代谢组学;同时测定;手性
Key words:
基金项目:中央高校基本科研业务费专项资金项目(21612343);;
2012年广东高校优秀青年创新人才培育计划(育苗工程)项目(2012LYM_0020)
中文摘要
目的研究D-酸性氨基酸的代谢组学,建立同时分离和定量体内样品中酸性氨基酸D、L体的离线二维HPLC法。方法利用硅胶基质ODS整体柱为第一维分离色谱柱,流动相为乙腈-三氟醋酸-水(9∶0.05∶91);Chiralpak QD-1-AX色谱柱为第二维手性分离柱,流动相为含10 mmol·L-1柠檬酸的甲醇-乙腈(50∶50),荧光法检测,离线手性2D-HPLC分析系统。结果两对对映体能同时得到良好地分离(Rs>2.50);各异构体的定量线性范围均为0.005~5 pmol(r>0.9998),检出限和定量限分别为2、5 fmol,回收率均在98%~105%,日内日间精密度的RSD均
参考文献
[1]Hamase K,Morikawa A,Zaitsu K.D-Amino acids in mammals and their diagnostic value[J].J Chromatogr B,2002,781(1):73-91.
[2]Brückner H,Schieber A.Ascertainment of D-amino acids in germ-free,gnotobiotic and normal laboratory rats[J].Biomed Chromatogr,2001,15(4):257-262.
[3]Huang AS,Beigneux A,Weil ZM,et al.D-aspartate regulates melanocortin formation and function:Behavioral alterations in Daspartate oxidase-deficient mice[J].J Neurosci,2006,26(10):2814-2819.
[4]Huang Y,Shi M,Zhao S.Quantification of D-Asp and D-Glu in rat brain and human cerebrospinal fluid by microchip electrophoresis[J].J Sep Sci,2009,32(17):3001-3006.
[5]Thorsén G,Bergquist J.Chiral separation of amino acids in biological fluids by micellar electrokinetic chromatography with laserinduced fluorescence detection[J].J Chromatogr B,2000,745(2):389-397.
[6]Fujii N,Kaji Y,Fujii N.D-Amino acids in aged proteins:Analysis and biological relevance[J].J Chromatogr B,2011,879(29):3141-3147.
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