Performance evaluation of long monolithic silica capillary columns in gradient liquid chromatography using peptide mixtures

• Published in: Journal of Chromatography A 2011-05, Vol.1218 (21), p.3360-3366
• Main Authors: Eghbali, Hamed, Sandra, Koen, Detobel, Frederik, Lynen, Frederic, Nakanishi, Kazuki, Sandra, Pat, Desmet, Gert
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Analytical, structural and metabolic biochemistry; Bed homogeneity; Biological and medical sciences; Chemistry; Chromatographic methods and physical methods associated with chromatography; Exact sciences and technology; flow resistance; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Gradient liquid chromatography; liquid chromatography; Long capillary column; Other chromatographic methods; Peak capacity; Proteins; silica; Silica monolith
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2010.10.045

A systematic study is reported on the performance of long monolithic capillary columns in gradient mode. Using a commercial nano-LC system, reversed-phase peptide separations obtained through UV-detection were conducted. The chromatographic performance, in terms of conditional peak capacity and peak productivity, was investigated for different gradient times (varying between 90 and 1320 min) and different column lengths (0.25, 1, 2 and 4 m) all originating from a single 4 m long column. Peak capacities reaching values up to n = 10 3 were measured in case of the 4 m long column demonstrating the high potential of these long monoliths for the analysis of complex biological mixtures, amongst others. In addition, it was found that the different column fragments displayed similar flow resistance as well as consistent chromatographic performance in accordance with chromatographic theory indicating that the chromatographic bed of the original 4 m long column possessed a structural homogeneity over its entire length.