Chiral separation of fluorescamine-labeled amino acids using microfabricated capillary electrophoresis devices for extraterrestrial exploration

被引:81
作者
Skelley, AM [1 ]
Mathies, RA [1 ]
机构
[1] Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA
基金
美国国家航空航天局;
关键词
enantiomer separation; extraterrestrial life detection; mars exploration; chip technology; derivatization; electrophoresis; amino acids; fluorescamine;
D O I
10.1016/j.chroma.2003.08.096
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Chiral separations of fluorescamine-labeled amino acids are characterized and optimized on a microfabricated capillary electrophoresis (CE) device. A standard mixture of acidic and neutral amino acids is labeled with fluorescamine in less than 5 min and the hydroxypropyl-beta-cyclodextrin (HPbetaCD) concentration, temperature, and pH are optimized (15 mM HPbetaCD, 6degreesC, pH < 9) to achieve high-quality and low background chiral separations in less than 200 s. All four stereoisomers formed in the labeling reaction of the chiral dye with the chiral amino acids are typically resolved. At pH > 9, isomerization of the dye chiral center is observed that occurs on the time scale of the chip separation. Typical limits of detection are similar to50 nM. These results demonstrate the feasibility of combining fluorescamine labeling of amino acids with microfabricated CE devices to develop low-volume, high-sensitivity apparatus and methods for extraterrestrial exploration. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:191 / 199
页数:9
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