Online, comprehensive two-dimensional liquid chromatography (2D-LC) has become an attractive option for the analysis of complex samples of relevance in various fields (e.g., environmental, food, biology, and polymer sciences). In general, the second-dimension (2D) separation plays a more crucial role than the first (1D) in the total performance of LC × LC systems. Speed and efficiency of the 2D separation are of primary importance. These challenges are amplified in the case of protein separations where mass transfer limitations effect elution kinetics. The research presented here leverages the developments of capillary-channeled polymer (C-CP) fiber stationary phases directed at high-throughput and rapid protein separations, evaluating them as 2D material in comprehensive reversed-phase × reversed-phase (RP × RP) 2D-LC versus conventional, packed-bed columns as 2D. The ability to operate fiber columns at high linear velocities (> 75 mm s-1 at < 1000 psi) without sacrifice in resolution is the first step towards potential 2D implementation, alleviating the need for ultrahigh-pressure pumping (i.e., UHPLC) in that stage. What must be realized are fast elution gradients and rapid re-equilibration to minimize cycling times (modulation periods). Different flow rates and gradient programs were evaluated. Rapid elution allows the use of shorter shift-gradient windows. At the optimized gradient recycling time on this instrument, 36 s, there is no significant degradation in peak capacity. Finally, a graphic comparison of the separation efficiency between seven commercial reversed-phase columns and the polypropylene C-CP fiber column is presented for a synthetic, eleven-protein mixture using the recommended operating conditions for each of the 2D columns. The kinetic advantages of the fiber columns are clearly demonstrated.

中文翻译：

---

聚丙烯毛细管通道聚合物纤维色谱柱是二维的完整蛋白质混合物的全面二维RP×RP分析中的第二维。

在线，全面的二维液相色谱（2D-LC）已成为分析各个领域（例如，环境，食品，生物学和高分子科学）中相关的复杂样品的一种有吸引力的选择。通常，在LC×LC系统的整体性能中，第二维（2D）分离比第一维（1D）扮演更重要的角色。2D分离的速度和效率至关重要。在蛋白质分离的情况下，这些挑战被放大，传质限制会影响洗脱动力学。本文介绍的研究利用了针对高通量和快速蛋白质分离的毛细管聚合物（C-CP）纤维固定相的发展，在综合反相×反相（RP×RP）2D中将它们评估为2D材料。 -LC与传统的 填充床色谱柱为2D。在不牺牲分辨率的情况下，以高线性速度（> 75 mm s-1在<1000 psi下）操作纤维柱的能力是实现潜在2D的第一步，从而减轻了对超高压泵送（即UHPLC）的需求阶段。必须实现快速洗脱梯度和快速重新平衡，以最大程度地减少循环时间（调制周期）。评价了不同的流速和梯度程序。快速洗脱允许使用更短的位移梯度窗口。在此仪器上最佳的梯度回收时间（36 s）下，峰容量没有明显下降。最后，对合成的七种商用反相色谱柱和聚丙烯C-CP纤维色谱柱的分离效率进行了图形比较。11种蛋白质混合物，使用建议的2D色谱柱操作条件。纤维柱的动力学优势已得到明确证明。