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Published March 15, 2000 | public
Journal Article

Quasi‐linear gradients for capillary liquid chromatography with mass and tandem mass spectrometry

Abstract

Gradient elution, capillary liquid chromatography mass spectrometry was performed with linear, static gradients constructed by laminar flowing ten, 1.5 mu L volume steps of decreasing organic concentration into tubing of small internal diameter. Sample loading, gradient formation, and sample elution were accomplished entirely by means of a commercially available micro-autosampler and single-syringe drive pump. The procedure was simple, fast, stable, and reproducible. Essentially linear gradients were produced without the use of additional valves, mixers, pumps or software. It took less than 10 minutes to form a gradient and less than 30 minutes to construct the set of individual buffer vials. The gradients were shown to be stable to storage. One hour after forming, peak retention times were reproduced to +/-0.5%. Long-term retention time reproducibility was found to vary by +/-2%, Chromatographic resolution was comparable or superior to that obtained by gradient elution with conventional dynamic mixing and split how, The procedure was adapted with a 'peak parking' method which extended the time for generating peptide fragmentation data up to 10 minutes per peptide with the triple quadruple mass spectrometer, Using this technique, collision data were collected at the 25 femtomole level on nine of ten tryptic peptides in a single run.

Additional Information

© 2000 John Wiley & Sons, Ltd. Issue online 15 March 2000; version of record online 15 March 2000; manuscript accepted 05 January 2000; manuscript received 03 January 2000; first published 15 March 2000. Funding for this research was provided by funds from The Caltech Beckman Institute. The authors thank the City of Hope, Department of Immunology, Protein Core Laboratory for providing the amino acid analyses.

Additional details

Created:
August 19, 2023
Modified:
October 18, 2023