Enhanced Amine and Amino Acid Analysis Using Pacific Blue and the Mars Organic Analyzer Microchip Capillary Electrophoresis System
Enhanced Amine and Amino Acid Analysis Using Pacific Blue and the Mars Organic Analyzer Microchip Capillary Electrophoresis System
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DOI：
10.1021/ac8023334
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发表时间：
2009-04-01
影响因子：
7.4
通讯作者：
Mathies, Richard A.
中科院分区：
文献类型：
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作者：
Chiesl, Thomas N.;Chu, Wai K.;Mathies, Richard A.
采用荧光胺反应探针太平洋蓝琥珀酰酰酯（PB）在火星有机分析仪（MOA）上通过微芯片毛细管电泳检测微量胺和氨基酸。与荧光胺衍生化相比，PB的光谱和化学性质提供了200倍的灵敏度和改进的分辨率。通过交叉注射和PB标记，MOA检测到的氨基酸浓度低至75 pM（万亿分之一）。胶束电动色谱法（MEKC）通过其疏水性来分离铅标记的氨基酸。优化的MEKC条件（45 mM CHAPSO, pH 6, 5℃）有效分离胺和25种氨基酸，对映体分辨率为丙氨酸、丝氨酸和瓜氨酸。来自智利阿塔卡马沙漠云盖山地区和默奇森陨石的样本使用这两种技术成功地进行了分析，氨基酸含量在十亿分之一的范围内。在Murchison样品中检测到非生物氨基酸，如β -丙氨酸和epsilon-氨基戊酸，以及几种中性和酸性氨基酸。阿塔卡马沙漠样品被发现含有同手性l -丙氨酸和l -丝氨酸，表明存在现存或最近灭绝的生命。
The fluorescent amine reactive probe Pacific Blue succinimidyl ester (PB) is used for the detection of trace amounts of amines and amino acids by microchip capillary electrophoresis on the Mars Organic Analyzer (MOA). The spectral and chemical properties of PB provide a 200-fold increase in sensitivity and improved resolution compared to fluorescamine derivatization. With the use of cross injection and PB labeling, the MOA detected amino acids at concentrations as low as 75 pM (sub-parts-per-trillion). Micellar electrokinetic chromatograpy (MEKC) which separates PB-labeled amino acids by their hydro-phobicity is also demonstrated. The optimized MEKC conditions (45 mM CHAPSO, pH 6 at 5 degrees C) effectively separated amines and 25 amino acids with enantiomeric resolution of alanine, serine, and citrulline. Samples from the Yungay Hills region in the Atacama Desert, Chile and from the Murchison meteorite are successfully analyzed using both techniques, and amino acids are found in the parts-per-billion range. Abiotic amino acids such as beta-alanine and epsilon-aminocaprioc acid are detected along with several neutral and acidic amino acids in the Murchison sample. The Atacama Desert sample is found to contain homochiral L-alanine and L-serine indicating the presence of extant or recently extinct life.