Testing and Quality Control of a Reversed-Phase Column Packing

Variations in separation factor from lot-to-lot were measured on 24 different manufacturer's lots of Vydac TP-201, a commercial reversed-phase liquid chromatography column packing material. Three pairs of test compounds were used in acetonitrile-water mobile phases. It was found that the pairs anthracene/phenanthrene and androstenedione/testosterone showed substantial lot-to-lot variations in separation factors and these varied largely independently. In contrast, the pair biphenyl/naphthalene showed lot-to-lot variations that were too small to measure easily. Separation factors for the three sample pairs for the Vydac material were compared with separation factors measured on 8 other commercially available C18 packing materials. These materials differed widely from each other in separation factors for anthracene/phenanthrene and androstenedione/testosterone, but all had similar separation factors for biphenyl/naphthalene. Quality control ranges that were established for separation factors and capacity ratios of lots to be used for packing columns are presented.