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Fast enantiomeric separation of amino acids using liquid chromatography/mass spectrometry on a chiral crown ether stationary phase.
Journal of Bioscience and Bioengineering ( IF 2.8 ) Pub Date : 2020-06-29 , DOI: 10.1016/j.jbiosc.2020.05.007
Kohei Yoshikawa 1 , Masahiro Furuno 1 , Nobuo Tanaka 1 , Eiichiro Fukusaki 1
Affiliation  

Fast enantiomeric separation of amino acids was studied by liquid chromatography/mass spectrometry (LC/MS) on a chiral crown ether stationary phase. A chiral crown ether bonded silica column (3 mm internal diameter (i.d.), 5 cm long) packed with 3 μm particles was employed instead of a 15 cm column packed with 5 μm particles used in our previous study. In addition, the extra-column variance, becoming more serious for smaller columns, was reduced by replacing 0.127 mm i.d. post-column tubes with shorter, smaller-diameter (0.0635 mm i.d.) tubes. The results demonstrated the benefits of using shorter columns packed with smaller particles and the reduction of the extra-column band broadening for fast enantiomeric separation. Finally, the enantiomeric separation of 18 pairs of proteinogenic amino acids was achieved within 2 min with a resolution (Rs) > 1.5 for each pair using an isocratic mobile phase of acetonitrile/water/trifluoroacetic acid (ACN/W/TFA) = 96/4/0.5, and a flow rate 1.2 mL/min at 30°C. This is the highest throughput method for simultaneous chiral separation of all proteinogenic amino acids except proline to date.



中文翻译:

使用液相色谱/质谱在手性冠醚固定相上快速对映体分离氨基酸。

通过液相色谱/质谱(LC / MS)在手性冠醚固定相上研究了氨基酸的快速对映体分离。使用填充有3μm颗粒的手性冠醚键合硅胶柱(内径3毫米,内径5厘米长)填充了3μm颗粒,而不是我们先前研究中使用的填充有5μm颗粒的15 cm柱。此外,通过用更短,更小直径(0.0635 mm内径)的管子代替0.127 mm内径的柱后管,可以减小对于较小的色谱柱而言更为严重的柱外差异。结果表明,使用较短的色谱柱填充较小的颗粒,并减少了色谱柱外谱带展宽,以实现快速的对映体分离。最后,使用乙腈/水/三氟乙酸的等度流动相(ACN / W / TFA)= 96/4 /,在2分钟内以18对成对氨基酸的对映体分离,每对的分离度(Rs)> 1.5 0.5,在30°C下的流速为1.2 mL / min。这是迄今为止除脯氨酸外的所有蛋白质氨基酸同时进行手性分离的最高通量方法。

更新日期:2020-06-29
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