Enhanced sensitivity using a direct flow capillary hplc/ms system with nanoflow capability

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Cozette M. Cuppett;Jeffrey Holyoke;Steven Cohen
ASMS 2000, Long Beach, CA; June 11-15
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Bovine Cytochrome c
XTerra C18 N/A N/A N/A N/A Symmetry® C18 5 µm N/A 0.180 mm 150 mm
The need for high sensitivity separation in sample limited applications has driven the development of system that couple capillary LC with MS. To increase sensitivity, the trend has been to scale down flow rates and incorporate nanoflow ESI interfaces on triple quadrupole and Q-TOF mass spectrometers. Typically, the low flow rates are achieved by performing a precolumn flow split. This precolumn split makes the flow through the column susceptible to rate variations due to backpressure changes with column age and this can greatly affect separation reproducibility. Using an integrated trapping configuration on a capillary HPLC system, a tryptic digest of bovine cytochrome c was loaded onto a focusing column at 20 ul/min, and then eluted through either a 75 or 180 um column with flow rates ranging from 400 nl/min to 2 ul/min. Detection of the separation was performed using micro flow and nanoflow electrospray interfaces on a single quadrupole mass spectrometer.

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