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Capillary liquid chromatography fraction collection and postcolumn reaction using segmented flow microfluidics
Author(s) -
Nie Jing,
Kennedy Robert T.
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201300725
Subject(s) - chromatography , chemistry , fraction (chemistry) , microscale chemistry , derivatization , capillary action , volumetric flow rate , microfluidics , resolution (logic) , reagent , analytical chemistry (journal) , mass spectrometry , nanotechnology , materials science , mathematics education , mathematics , physics , quantum mechanics , artificial intelligence , computer science , composite material
A challenge for capillary LC (c LC ) is fraction collection and the manipulation of fractions from microscale columns. An emerging approach is the use of segmented flow or droplet technology to perform such tasks. In this work, a fraction collection and postcolumn reaction system based on segmented flow was developed for the gradient c LC of proteins. In the system, column effluent and immiscible oil are pumped into separate arms of a tee resulting in regular fractions of effluent segmented by oil. Fractions were generated at 1 Hz corresponding to 5 nL volumes. The fraction collection rate was high enough to generate over 30 fractions per peak and preserve chromatographic resolution achieved for a five‐protein test mixture. The resulting fractions could be stored and subsequently derivatized for fluorescence detection by pumping them into a second tee where naphthalene dicarboxyaldehyde, a fluorogenic reagent, was pumped into a second arm and added to each fraction. Proteins were derivatized within the droplets enabling postcolumn fluorescence detection of the proteins. The experiments demonstrate that fraction collection from c LC by segmented flow can be extended to proteins. Further, they illustrate a potential workflow for protein analysis based on postcolumn derivatization for fluorescence detection.