Ion Mobility Mass Spectrometry of Peptide Ions: Effects of Drift Gas and Calibration Strategies

TLDR

Ion mobility mass spectrometry’s utility for peptide assignment is limited by the variability of pressure, gas composition, temperature, and separation mode, which hampers rapid extraction of accurate shape parameters. The authors measured collision cross sections for 113 peptide ions (masses 231–2969 Da) in both He and N₂ using a low‑pressure, ambient‑temperature drift cell with RF confinement, providing a broad calibration set. These measurements quantified errors in traveling‑wave Ω determinations and in estimating He Ω from N₂ drift times, and demonstrate that calibrated Ω can be rapidly obtained to enhance proteomic assignment specificity.

Abstract

One difficulty in using ion mobility (IM) mass spectrometry (MS) to improve the specificity of peptide ion assignments is that IM separations are performed using a range of pressures, gas compositions, temperatures, and modes of separation, which makes it challenging to rapidly extract accurate shape parameters. We report collision cross section values (Ω) in both He and N(2) gases for 113 peptide ions determined directly from drift times measured in a low-pressure, ambient temperature drift cell with radio-frequency (rf) ion confinement. These peptide ions have masses ranging from 231 to 2969 Da, Ω(He) of 89-616 Å(2), and Ω(N(2)) of 151-801 Å(2); thus, they are ideal for calibrating results from proteomics experiments. These results were used to quantify the errors associated with traveling-wave Ω measurements of peptide ions and the errors concomitant with using drift times measured in N(2) gas to estimate Ω(He). More broadly, these results enable the rapid and accurate determination of calibrated Ω for peptide ions, which could be used as an additional parameter to increase the specificity of assignments in proteomics experiments.