RationaleTandem‐ion mobility spectrometry/mass spectrometry methods have recently gained traction for the structural characterization of proteins and protein complexes. However, ion activation techniques currently coupled with tandem‐ion mobility spectrometry/mass spectrometry methods are limited in their ability to characterize structures of proteins and protein complexes. MethodsHere, we describe the coupling of the separation capabilities of tandem‐trapped ion mobility spectrometry/mass spectrometry (tTIMS/MS) with the dissociation capabilities of ultraviolet photodissociation (UVPD) for protein structure analysis. ResultsWe establish the feasibility of dissociating intact proteins by UV irradiation at 213 nm between the two TIMS devices in tTIMS/MS and at pressure conditions compatible with ion mobility spectrometry (2–3 mbar). We validate that the fragments produced by UVPD under these conditions result from a radical‐based mechanism in accordance with prior literature on UVPD. The data suggest stabilization of fragment ions produced from UVPD by collisional cooling due to the elevated pressures used here (“UVnoD2”), which otherwise do not survive to detection. The data account for a sequence coverage for the protein ubiquitin comparable to recent reports, demonstrating the analytical utility of our instrument in mobility‐separating fragment ions produced from UVPD. ConclusionsThe data demonstrate that UVPD carried out at elevated pressures of 2–3 mbar yields extensive fragment ions rich in information about the protein and that their exhaustive analysis requires IMS separation post‐UVPD. Therefore, because UVPD and tTIMS/MS each have been shown to be valuable techniques on their own merit in proteomics, our contribution here underscores the potential of combining tTIMS/MS with UVPD for structural proteomics.
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Detection of firearm discharge residue from skin swabs using trapped ion mobility spectrometry coupled to mass spectrometry
In the present work, a novel workflow for the detection of both elemental and organic constituents of the firearm discharge residue from skin swabs was developed using trapped ion mobility spectrometry coupled to mass spectrometry. The small sample size (<10 μL), high specificity and short analysis time (few min) permits the detection of inorganic residues (IGSR; inorganic gunshot residues) and organic residues (OGSR) from one sample and in a single analysis. The analytical method is based on the simultaneous extraction of inorganic and organic species assisted by the formation organometallic complexes ( e.g. , 15–5 crown ethers for the sequestering of metals and nitrate species), followed by fast, post-ionization, high resolution mobility ( R IMS ∼ 150–250) and mass separations ( R MS ∼ 20–40k) with isotopic pattern recognition. The analytical performance is illustrated as a proof of concept for the case of the simultaneous detection of Ba +2 , Pb +2 , Cu + , K + , NO 3 − , diphenylamine (DPA), ethyl centralite (EC) and 2,4 dinitrotoluene (DNT) in positive and negative nESI-TIMS-MS modes. Candidate structures are proposed and collisional cross sections are reported for all organic and organometallic species of interest.
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- Award ID(s):
- 1654274
- PAR ID:
- 10078816
- Date Published:
- Journal Name:
- Analytical Methods
- Volume:
- 10
- Issue:
- 35
- ISSN:
- 1759-9660
- Page Range / eLocation ID:
- 4219 to 4224
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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