Mass measurement errors of Fourier-transform mass spectrometry (FTMS): distribution, recalibration, and application

J Proteome Res. 2009 Feb;8(2):849-59. doi: 10.1021/pr8005588.

Abstract

The hybrid linear trap quadrupole Fourier-transform (LTQ-FT) ion cyclotron resonance mass spectrometer, an instrument with high accuracy and resolution, is widely used in the identification and quantification of peptides and proteins. However, time-dependent errors in the system may lead to deterioration of the accuracy of these instruments, negatively influencing the determination of the mass error tolerance (MET) in database searches. Here, a comprehensive discussion of LTQ/FT precursor ion mass error is provided. On the basis of an investigation of the mass error distribution, we propose an improved recalibration formula and introduce a new tool, FTDR (Fourier-transform data recalibration), that employs a graphic user interface (GUI) for automatic calibration. It was found that the calibration could adjust the mass error distribution to more closely approximate a normal distribution and reduce the standard deviation (SD). Consequently, we present a new strategy, LDSF (Large MET database search and small MET filtration), for database search MET specification and validation of database search results. As the name implies, a large-MET database search is conducted and the search results are then filtered using the statistical MET estimated from high-confidence results. By applying this strategy to a standard protein data set and a complex data set, we demonstrate the LDSF can significantly improve the sensitivity of the result validation procedure.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calibration
  • Databases, Protein
  • Fourier Analysis*
  • Humans
  • Mass Spectrometry* / instrumentation
  • Mass Spectrometry* / methods
  • Peptides / analysis
  • Proteomics / instrumentation*
  • Proteomics / methods
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Peptides