Gritti Fabrice, Guiochon Georges
Department of Chemistry, University of Tennessee, Knoxville, TN 37996-1600, USA.
J Chromatogr A. 2007 Sep 28;1166(1-2):30-46. doi: 10.1016/j.chroma.2007.06.064. Epub 2007 Jul 5.
We measured and compared the characteristics and performance of columns packed with particles of five different C(18)-bonded silica, 3 and 5 microm Luna, 3 microm Atlantis, 3.5 microm Zorbax, and 2.7 microm Halo. The average particle size of each material was derived from the SEM pictures of 200 individual particles. These pictures contrast the irregular morphology of the external surface of the Zorbax and Halo particles and the smooth surface of the Luna and Atlantis particles. In a wide range of mobile phase velocities (from 0.010 to 3 mL/min) and at ambient temperature, we measured the first and second central moments of the peaks of naphthalene, insulin, and bovine serum albumin (BSA). These moments were corrected for the contributions of the extra-column volumes to calculate the reduced HETPs. The C-terms of naphthalene and insulin are largest for the Halo and Zorbax materials and the A-term smallest for the Halo-packed column. The Halo column performs the best for the low molecular weight compound naphthalene (minimum reduced HETP, 1.4) but is not as good as the Atlantis or Luna columns for the large molecular weight compound insulin. The Zorbax column is the least efficient column because of its large C-term. The lowest sample diffusivity through these particles, alone, does not account for the results. It is most likely that the roughness of the external surface of the Halo and Zorbax particles limit the performance of these columns at high flow rates generating an unusually high film mass transfer resistance.
我们测量并比较了填充有五种不同C(18)键合硅胶颗粒的色谱柱的特性和性能,分别是3微米和5微米的Luna、3微米的Atlantis、3.5微米的Zorbax以及2.7微米的Halo。每种材料的平均粒径由200个单个颗粒的扫描电子显微镜图片得出。这些图片对比了Zorbax和Halo颗粒外表面不规则的形态以及Luna和Atlantis颗粒光滑的表面。在较宽的流动相流速范围(从0.010至3 mL/分钟)以及环境温度下,我们测量了萘、胰岛素和牛血清白蛋白(BSA)峰的一阶和二阶中心矩。对这些矩进行了柱外体积贡献的校正,以计算折合理论塔板高度。对于萘和胰岛素,Halo和Zorbax材料的C项最大,而对于填充Halo的色谱柱,A项最小。Halo色谱柱对低分子量化合物萘的表现最佳(折合理论塔板高度最小,为1.4),但对于大分子化合物胰岛素,其表现不如Atlantis或Luna色谱柱。由于其较大的C项,Zorbax色谱柱是效率最低的色谱柱。仅颗粒中最低的样品扩散率并不能解释这些结果。很可能是Halo和Zorbax颗粒外表面的粗糙度限制了这些色谱柱在高流速下的性能,从而产生了异常高的液膜传质阻力。