期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1469, 期 -, 页码 60-67出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2016.09.053
关键词
UHPLC; Capillary chromatography; LC-MS; Proteomics; Peptides
资金
- Waters Corporation (Milford, MA)
- National Institute of Health [5U24DK097153]
- National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) [1R01DK101473-01A1]
Commercial chromatographic instrumentation for bottom-up proteomics is often inadequate to resolve the number of peptides in many samples. This has inspired a number of complex approaches to increase peak capacity, including various multidimensional approaches, and reliance on advancements in mass spectrometry. One-dimensional reversed phase separations are limited by the pressure capabilities of commercial instruments and prevent the realization of greater separation power in terms of speed and resolution inherent to smaller sorbents and ultrahigh pressure liquid chromatography. Many applications with complex samples could benefit from the increased separation performance of long capillary columns packed with sub-2 mu m sorbents. Here, we introduce a system that operates at a constant pressure and is capable of separations at pressures up to 45 kpsi. The system consists of a commercially available capillary liquid chromatography instrument, for sample management and gradient creation, and is modified with a storage loop and isolated pneumatic amplifier pump for elevated separation pressure. The system's performance is assessed with a complex peptide mixture and a range of microcapillary columns packed with sub-2 mu m C18 particles. (C) 2016 Elsevier B.V. All rights reserved.
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