Evaluation of nano-liquid chromatography-tandem mass spectrometry in a column switching setup for the absolute quantification of peptides in the picomolar range.
A standard nanospray-liquid chromatography-tandem mass spectrometry system in a column switching setup for the absolute quantification of leucine-enkephalin was evaluated. Analytes were loaded on a C18 trapping column and back-flushed in the 75 microm analytical column. Quantification was performed with a triple quadrupole instrument. Validation results show that it is feasible, with a conventional nano-LC system in the column switching setup, to quantify peptides as low as 500 amol on column (50 pmol/L). Weighted linear regression analysis proves a good linearity in a dynamic range of almost three orders of magnitude. Nevertheless, robustness remains a key issue in nano-LC-MS/MS.
Sinnaeve
Bart A
BA
Laboratoria voor Medische Biochemie en voor Klinische Analyse, Universiteit Gent, Harelbekestraat 72, B-9000 Gent, Belgium.
Van Bocxlaer
Jan F
JF
D002853:Chromatography, Liquid / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D036103:Nanotechnology; D010455:Peptides / Q000032:analysis*
D010455:Peptides