Kiyonami R, Ravnsborg C, Zabrouskov V.
ASMS 2012 Poster
Purpose: Evaluate the performance of a new nanospray ion source and nanoflow
column assemblies that incorporate a nanoflow column, column heater, high-voltage
electrode, and emitter into an integrated unit. Key aspects of chromatographic
performance are evaluated: chromatographic resolution, retention-time
reproducibility, sample loading capacity, and column robustness.
Methods: A prototype nanospray source was installed on the mass spectrometer
and connected to a nanoLC pump. Column assemblies were tested to evaluate
column-to-column and run-to-run reproducibility. Complex peptide mixtures and a
simple digest mixture were used for the evaluation. The retention-time reproducibility,
peak shape, resolution and peak capacity, in different temperature ranges, were
evaluated. In order to test the device flexibility, several flow rates from 150 nL/min to
500 nL/min were used.
Results: A high spray stability was achieved over all LC runs. The chromatographic
resolution, sensitivity, and reproducibility obtained matched other state-of-the-art
data without need for adjustments, intervention or other actions beyond simply
inserting the column/sprayer assembly into the nanospray source. More than 1000
proteins were identified with increased component detection in the complex protein
digest mixtures. The column-to-column retention-time reproducibility for the targeted
peptides in the simple digest mixture was excellent.
Thermo Fisher Scientific