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Application of Fused-Core Particle Column in Two Dimensional Reversed Phase - Reversed Phase LC/MS Analysis of Biological Samples. Impact of Extra-Column Volume

Eduard Rogatsky, Shaynah Browne, Bing Na Zheng and Daniel T. Stein

We demonstrate successful applicability of fused–core column in analysis of biological samples (diluted urine and SPE–pre-purified plasma or serum) using 2 dimensional chromatography. Implementation of fused-core material in the second dimension of 2D chromatographic platform allowed improving S/N above 4 fold in comparison to traditional porous C18 column. Sample load onto the first dimension (5 μm) eliminates limitations of sample volume to be injected. The main limitation factor of the fused-core material–low loading capacity was overcome by using 2D chromatography. We emphasize importance of reduction of post-column (extra column) volume in LC/MS applications utilizing fused-core columns.