Banes A J, Link G W, Snyder L R
J Chromatogr. 1985 Jun 19;326:419-31. doi: 10.1016/s0021-9673(01)87467-7.
A total of 10 reversed-phase columns obtained from different suppliers were evaluated for their ability to separate typical peptide mixtures: tryptic digests of cytochrome c, lysozyme and collagen. Each column was tested in a standard gradient elution using phosphate buffer (pH 2.85) as the A-solvent and acetonitrile as the B-solvent. Some differences in band spacing for the various peptides were observed from column to column. More important differences were observed in the ability of these various columns to provide narrow peaks and good resolution, as measured by column peak capacity. These differences in column peak capacity were related to differences in column dimensions and particle size; it also appeared that C8 bonded phases were somewhat more efficient than were C18 phases. The recovery of cytochrome c digest from four of the columns was also assessed. Summed peak areas were proportional to sample size, as the latter was varied from 1 to 100 micrograms. The apparent recovery from each column did not vary by more than +/- 5%. From this it was concluded that these columns gave essentially quantitative recovery for this particular sample. Limited data on column life were obtained for some of these columns.
对从不同供应商处获得的10根反相柱进行了评估，以考察它们分离典型肽混合物的能力，这些混合物包括细胞色素c、溶菌酶和胶原蛋白的胰蛋白酶消化产物。每根柱子都在标准梯度洗脱条件下进行测试，使用磷酸盐缓冲液（pH 2.85）作为A溶剂，乙腈作为B溶剂。不同柱子之间观察到了各种肽的谱带间距存在一些差异。通过柱峰容量衡量，这些不同柱子在提供窄峰和良好分离度的能力方面观察到了更重要的差异。柱峰容量的这些差异与柱尺寸和粒径的差异有关；似乎C8键合相在某种程度上比C18相更高效。还评估了其中四根柱子对细胞色素c消化产物的回收率。当样品量从1微克变化到100微克时，峰面积总和与样品量成正比。每根柱子的表观回收率变化不超过±5%。由此得出结论，对于该特定样品，这些柱子基本上实现了定量回收。获得了其中一些柱子有限的柱寿命数据。
