Schure Mark R, Moran Robert E
Theoretical Separation Science Laboratory, Kroungold Analytical, Inc., 1299 Butler Pike, Blue Bell, PA, 19422 USA.
Advanced Materials Technology, Inc., 3521 Silverside Road, Suite 1-K, Quillen Building, Wilmington, DE, 19810, USA.
J Chromatogr A. 2017 Jan 13;1480:11-19. doi: 10.1016/j.chroma.2016.12.016. Epub 2016 Dec 9.
A comparison is made using size-exclusion chromatography (SEC) of synthetic polymers between fully porous particles (FPPs) and superficially porous particles (SPPs) with similar particle diameters, pore sizes and equal flow rates. Polystyrene molecular weight standards with a mobile phase of tetrahydrofuran are utilized for all measurements conducted with standard HPLC equipment. Although it is traditionally thought that larger pore volume is thermodynamically advantageous in SEC for better separations, SPPs have kinetic advantages and these will be shown to compensate for the loss in pore volume compared to FPPs. The comparison metrics include the elution range (smaller with SPPs), the plate count (larger for SPPs), the rate production of theoretical plates (larger for SPPs) and the specific resolution (larger with FPPs). Advantages to using SPPs for SEC are discussed such that similar separations can be conducted faster using SPPs. SEC using SPPs offers similar peak capacities to that using FPPs but with faster operation. This also suggests that SEC conducted in the second dimension of a two-dimensional liquid chromatograph may benefit with reduced run time and with equivalently reduced peak width making SPPs advantageous for sampling the first dimension by the second dimension separator. Additional advantages are discussed for biomolecules along with a discussion of optimization criteria for size-based separations.
使用尺寸排阻色谱法(SEC),对粒径、孔径相似且流速相同的全多孔颗粒(FPP)和表面多孔颗粒(SPP)之间的合成聚合物进行比较。所有测量均使用配备四氢呋喃流动相的聚苯乙烯分子量标准品,通过标准高效液相色谱设备进行。传统观点认为,在SEC中,更大的孔体积在热力学上有利于实现更好的分离,但表面多孔颗粒具有动力学优势,并且将证明这些优势能够弥补与全多孔颗粒相比孔体积的损失。比较指标包括洗脱范围(表面多孔颗粒更小)、塔板数(表面多孔颗粒更大)、理论塔板生成速率(表面多孔颗粒更大)和比分辨率(全多孔颗粒更大)。文中讨论了在SEC中使用表面多孔颗粒的优势,即使用表面多孔颗粒可以更快地进行类似的分离。使用表面多孔颗粒的SEC与使用全多孔颗粒的SEC具有相似的峰容量,但操作速度更快。这也表明,在二维液相色谱的第二维中进行的SEC可能会受益于运行时间的减少,以及等效的峰宽减小,这使得表面多孔颗粒有利于由第二维分离器对第一维进行进样。文中还讨论了生物分子的其他优势,以及基于尺寸分离的优化标准。