Orthogonality of separation in two-dimensional liquid chromatography
Orthogonality of separation in two-dimensional liquid chromatography
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DOI：
10.1021/ac050923i
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发表时间：
2005-10-01
影响因子：
7.4
通讯作者：
Gebler, JC
中科院分区：
文献类型：
--
作者：
Gilar, M;Olivova, P;Gebler, JC
二维液相色谱通常用于在串联质谱分析之前降低蛋白质组样品的复杂性。二维液相色谱性能取决于色谱尺寸和分离正交性的峰容量。许多 LC 模式对肽的峰容量和选择性尚不清楚，正交性的数学表征也不完善。因此，很难评估 2D-LC 在肽分离方面的性能。本文的目的是研究常见 LC 模式的选择性并识别具有有用正交性的 2DLC 系统。开发了正交描述的几何方法，并将其应用于实际峰值 2D-LC 容量的估计。选定的 LC 模式（包括各种 RP、SCX、SEC 和 HILIC）在 2D-LC 设置中组合。 SCX-RP、HILIG RP 和 RP-RP 2D 系统可提供合适的正交性。 RP-RP 系统（在两个 RP 分离维度上采用显着不同的 pH 值）具有所研究的 2D-LC 系统中最高的实际峰容量。
Two-dimensional liquid chromatography is often used to reduce the proteomic sample complexity prior to tandem mass spectrometry analysis. The 2D-LC performance depends on the peak capacity in both chromatographic dimensions, and separation orthogonality. The peak capacity and selectivity of many LC modes for peptides is not well known, and mathematical characterization for orthogonality is underdeveloped. Consequently, it is difficult to estimate the performance of 2D-LC for peptide separation. The goal of this paper was to investigate a selectivity of common LC modes and to identify the 2DLC systems with a useful orthogonality. A geometric approach for orthogonality description was developed and applied for estimation of a practical peak 2D-LC capacity. Selected LC modes including various RP, SCX, SEC, and HILIC were combined in 2D-LC setups. SCX-RP, HILIG RP, and RP-RP 2D systems were found to provide suitable orthogonality. The RP-RP system (employing significantly different pH in both RP separation dimensions) had the highest practical peak capacity of 2D-LC systems investigated.