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Ultrahigh-Pressure Reversed-Phase Capillary Liquid Chromatography: Isocratic and Gradient Elution Using Columns Packed with 1.0-μm Particles
Abstract
Fused-silica capillaries with inner diameters of 33 microns and lengths of 25-50 cm are slurry-packed with 1.0-micron nonporous octadecylsilane-modified (C18) silica spheres. These columns are used to perform ultrahigh-pressure reversed-phase liquid chromatographic analyses in both isocratic and gradient elution modes. Mobile-phase pressures as high as 5000 bar (72,000 psi) are applied to column inlets to generate more than 200,000 theoretical plates in 6 min (k' approximately 1) for small, organic analytes. Average capacity factors of analytes are found to increase linearly with applied pressure. An electrically driven constant-flow syringe pump capable of generating mobile-phase pressures as high as 9000 bar (130,000 psi) is described. This pump is used in conjunction with an exponential dilution method for the gradient separation of peptides from a tryptic digest on a 27-cm-long capillary packed with 1.0-micron particles. A peak capacity of 300 is demonstrated for a 30-min analysis.
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Cited by
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Nguyen, D. T. ‐., Guillarme, D., Rudaz, S., & Veuthey, J. (2006) · Journal of Separation Science · doi:10.1002/jssc.200600189 · Cited by 337
Ion mobility–mass spectrometry: a new paradigm for proteomics
McLean, J. A., Ruotolo, B. T., Gillig, K. J., & Russell, D. H. (2005) · International Journal of Mass Spectrometry · doi:10.1016/j.ijms.2004.10.003 · Cited by 309
Rapid and Deep Human Proteome Analysis by Single-dimension Shotgun Proteomics
Pirmoradian, M., Budamgunta, H., Chingin, K., Zhang, B., Astorga-Wells, J., & Zubarev, R. A. (2013) · Molecular & Cellular Proteomics · doi:10.1074/mcp.o113.028787 · Cited by 144
Metadata
- Authors
- MacNair, J. E., Patel, K. D., & Jorgenson, J. W.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1999-01-05
- Volume
- 71
- Issue
- 3
- Pages
- 700-708
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W2015538298
- Cited by
- 377
- References
- 0
- FWCI
- 8.74
- Type
- journal article
- Language
- en
- Open access
- closed
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- CoChat. (2026). Citation check: Ultrahigh-Pressure Reversed-Phase Capillary Liquid Chromatography: Isocratic and Gradient Elution Using Columns Packed with 1.0-μm Particles. https://app.cochat.ai/papers/10.1021/ac9807013
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- MacNair, J. E., Patel, K. D., & Jorgenson, J. W. (1999). Ultrahigh-Pressure Reversed-Phase Capillary Liquid Chromatography: Isocratic and Gradient Elution Using Columns Packed with 1.0-μm Particles. Analytical Chemistry. https://doi.org/10.1021/ac9807013
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