Cole S R, Dorsey J G
Department of Chemistry, University of Cincinnati, OH 45221-0172, USA.
Biomed Chromatogr. 1997 May-Jun;11(3):167-71. doi: 10.1002/(SICI)1099-0801(199705)11:3<167::AID-BMC673>3.0.CO;2-1.
While the choice of stationary phase, organic modifier, and gradient strength can have significant effects on biomolecule separations, mobile phase additives can also have a significant effect on the chromatographic selectivity, recovery, efficiency and resolution. Given the importance of stationary phase coverage, the beneficial, silanol-masking properties of amines, and the potential for selectivity modification through ion-pair interactions, cyclohexylamine was examined as a mobile phase additive and compared with triethylamine and trifluoroacetic acid. Greatly improved separation was possible when cyclohexylamine was used as compared with phosphate buffer, and cyclohexylamine did not require purification before use, while triethylamine required distillation before 'clean' chromatograms were obtained.
虽然固定相、有机改性剂和梯度强度的选择会对生物分子分离产生显著影响，但流动相添加剂也会对色谱选择性、回收率、效率和分离度产生显著影响。鉴于固定相覆盖的重要性、胺类物质有益的硅醇掩蔽特性以及通过离子对相互作用改变选择性的可能性，研究了环己胺作为流动相添加剂，并与三乙胺和三氟乙酸进行了比较。与磷酸盐缓冲液相比，使用环己胺时分离效果有了很大改善，并且环己胺在使用前无需纯化，而三乙胺在获得“干净”的色谱图之前需要蒸馏。
