Quantification of biomolecules by external electrospray ionization Fourier transform mass spectrometry.
Fourier transform mass spectrometry (FTMS) is well-known for its capabilities in structural characterization of molecules. Recent developments in radio frequency excitation, linearized trapping, and accumulation of ions generated from external sources have improved the potential of FTMS for quantitative analysis. Here, a commercial external electrospray ionization FTMS, employing a linearized ion trap (the Infinity Cell) and an ion accumulation procedure in which ions are deflected off-axis and injected into the trap, is evaluated as an analytical method for quantifying amino acids, peptides, and proteins. Linear response over approximately 2-3 orders of magnitude is observed for singly-charged ions with low coefficients of variation (generally < 10%), and the calibration curves generated can be used to quantify structurally similar analytes with < 4% relative error, as shown here for quantification of leucine enkephalin from curves generated by methionine enkephalin. Similar precision is obtained for multiply-charged lysozyme, but over only 1.5 orders of magnitude. Some m/z discrimination is observed as a function of trap accumulation potential for a two-component cytochrome c/lysozyme mixture. The results are promising because they suggest that quantification using liquid chromatography coupled to electrospray FTMS is possible.
Padley
H R
HR
Department of Chemistry, Natural Sciences and Mathematics Complex, State University of New York at Buffalo 14260-3000, USA.
Bashir
S
S
Wood
T D
TD
D002853:Chromatography, Liquid / Q000379:methods; D004743:Enkephalin, Leucine / Q000032:analysis*; D004744:Enkephalin, Methionine / Q000032:analysis*; D005583:Fourier Analysis; D013058:Mass Spectrometry / Q000379:methods*; D009113:Muramidase / Q000032:analysis*; D015203:Reproducibility of Results
D004744:Enkephalin, Methionine; D004743:Enkephalin, Leucine; D009113:Muramidase