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Quasi‐linear gradients for capillary liquid chromatography with mass and tandem mass spectrometry
Author(s) -
Zhou Jie,
Rusnak Felicia,
Colonius Tim,
Hathaway Gary M.
Publication year - 2000
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/(sici)1097-0231(20000331)14:6<432::aid-rcm886>3.0.co;2-t
Subject(s) - chemistry , chromatography , analytical chemistry (journal) , mass spectrometry , elution , reproducibility , capillary action , syringe driver , resolution (logic) , tandem mass spectrometry , gradient elution , analyte , high performance liquid chromatography , syringe , psychology , materials science , composite material , artificial intelligence , psychiatry , computer science
Gradient elution, capillary liquid chromatography mass spectrometry was performed with linear, static gradients constructed by laminar flowing ten, 1.5 µL 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 flow. 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. Copyright © 2000 John Wiley & Sons, Ltd.