Fast, comprehensive online two-dimensional high performance liquid chromatography through the use of high temperature ultra-fast gradient elution reversed-phase liquid chromatography
Fast, comprehensive online two-dimensional high performance liquid chromatography through the use of high temperature ultra-fast gradient elution reversed-phase liquid chromatography
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DOI：
10.1016/j.chroma.2006.04.058
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发表时间：
2006-07-28
影响因子：
4.1
通讯作者：
Carr, Peter W.
中科院分区：
文献类型：
--
作者：
Stoll, Dwight R.;Cohen, Jerry D.;Carr, Peter W.
介绍了一种基于超快速高温梯度洗脱反相色谱的高速、全面在线双梯度洗脱2DLC（LC × LC）新方法。将非常常规的梯度洗脱仪器和柱组装在一个系统中，该系统在25分钟内产生约900的总峰容量;这大致相当于一个峰/2秒。每个第二维梯度在21秒的循环时间内完成，并且对于一组26个吲哚-3-乙酸（IAA）衍生物测量的峰保留时间可再现至0.2秒。从第一维色谱柱洗脱的每个峰在其宽度上至少取样两次，因为第二维色谱柱上的相应峰出现在两个或三个连续的第二维色谱图中，这清楚地表明在第一维分离中获得的分离度几乎没有损失。应用于野生型和突变玉米幼苗的低分子量组分的分离表明在25分钟的时间尺度上存在约100个峰。快速、高温2DLC的分析潜力的令人信服的说明是显而易见的，在来自单个相当窄的第一维峰的单个第二维色谱图中清楚地存在9个不同的峰，以及2DLC的强大功能，以解决在测量与巨大峰值相邻的小峰值时固有的“分析动态范围”问题。(c)2006 Elsevier B. V.保留所有权利。
A new approach to high speed, comprehensive online dual gradient elution 2DLC (LC x LC) based on the use of ultra-fast, high temperature gradient elution reversed phase chromatography is described. Entirely conventional gradient elution instrumentation and columns are assembled in a system which develops a total peak capacity of about 900 in 25 min; this is equivalent to roughly one peak/2 s. Each second dimension gradient is done in a cycle time of 21 s and the peak retention times measured for a set of twenty six indole-3-acetic acid (IAA) derivatives are reproducible to 0.2 s. Each peak eluting from the first dimension column is sampled at least twice across its width, as the corresponding peaks on the second dimension column appear in two or three consecutive second dimension chromatograms, clearly indicating that there is little loss in the resolution gained in the first dimension separation. Application to the separation of the low molecular weight components of wild-type and mutant maize seedlings indicates the presence of about 100 peaks on a timescale of 25 min. Compelling illustrations of the analytical potential of fast, high temperature 2DLC are evident in the clear presence of nine distinct peaks in a single second dimension chromatogram from a single quite narrow first dimension peak, and the great power of 2DLC to solve the "analytic dynamic range" problem inherent in the measurement of small peaks that are neighbors to a gigantic peak. (c) 2006 Elsevier B.V. All rights reserved.