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Structural analysis of a compound despite the presence of an isobaric interference by using in‐source Collision Induced Dissociation and tandem mass spectrometry
Journal of Mass Spectrometry  (IF1.982),  Pub Date : 2020-12-21, DOI: 10.1002/jms.4698
Dany Jeanne Dit Fouque, Alicia Maroto, Antony Memboeuf

The presence of an isobaric contaminant can drastically affect MS and MS/MS patterns leading to erroneous structural and quantitative analysis, which is a real challenge in mass spectrometry. Herein, we demonstrate that MS and MS/MS structural analysis of a compound can be successfully performed despite the presence of an isobaric interference with as low as few millidaltons mass difference by using pseudo‐MS3. To this end, in‐source collisional excitation (in‐source CID) and the Survival Yield (SY) technique (energy‐resolved collision induced dissociation MS/MS) were performed on two different source geometries: a Z‐spray and an orthogonal spray (with a transfer capillary) ionization sources on two different mass spectrometers. By using soft ionization conditions, the SY curve for the mixture is a linear combination of the SY curves from the pure compounds demonstrating the presence of two components in the mixture. In the case of harsher ionization conditions, the SY curve of the mixture perfectly overlaps the SY curve from the pure analyte. This observation demonstrates the isobaric interference has been completely removed by in‐source CID fragmentation, independently of the source design, leaving then the analyte precursor ions only. Therefore, by measuring the MS spectrum in harsh ionization conditions and according to SY criterium, the compound of interest can be made free from isobaric interference paving the way for, for example, unequivocal HPLC‐MS as well as HPLC‐MS/MS structural and quantitative analysis despite the presence of a co‐eluting isobaric interference.