Article (Scientific journals)
Liquid chromatography setup-dependent artefactual methionine oxidation of peptides: the importance of an adapted quality control process
Baumans, France; Hanozin, Emeline; Baiwir, Dominique et al.
2021In Journal of Chromatography. A, 1654
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Keywords :
methionine oxidation; liquid chromatography; peak broadening; mass spectrometry; complex samples; stationary phases
Abstract :
[en] In both biologics quality control experiments and protein post-translational modification studies, the analytical system used is not supposed to bring any artefactual modifications which could impair the results. In this work, we investigated oxidation of methionine-containing peptides during reversed-phase (RP) chromatographic separation. We first used a synthetic methionine-containing peptide to evaluate this artefactual phenomenon and then considered more complex samples (i.e., plasma and HeLa protein digests). The methionine oxidation levels of the peptides were systematically assessed and compared for the long-term use of the analytical column, the sample trapping time, the gradient length, the sample load and the nature of the stationary phase (HSS T3 from Waters, YMC Triart C18 from YMC Europe GmbH and BEH130 C18 from Waters). In addition to the oxidation of methionine in solution, we observed on the HSS T3 and the BEH130 stationary phases an additional broad peak corresponding to an on-column oxidized species. Considering the HSS T3 phase, our results highlight that the on-column oxidation level significantly increases with the age of the analytical column and the gradient length and reaches 56 % when a 1-year-old column set is used with a 180 min-long LC method. These levels go to 0 % and 18 % for the YMC Triart C18 and the BEH130 C18 phases respectively. Interestingly, the on-column oxidation proportion decreases as the injected sample load increases suggesting the presence of a discrete number of oxidation sites within the stationary phase of the analytical column. Those findings observed in different laboratories using distinct set of columns, albeit to varying degrees, strengthen the need for a standard of methionine-containing peptide that could be used as a quality control to appraise the status of the liquid chromatographic columns.
Disciplines :
Chemistry
Author, co-author :
Baumans, France ;  Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Hanozin, Emeline ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Baiwir, Dominique  ;  Université de Liège - ULiège > GIGA Platforms
Decroo, Corentin;  University of Mons > Proteomics and Microbiology Laboratory
Wattiez, Ruddy;  University of Mons > Proteomics and Microbiology Laboratory
De Pauw, Edwin  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Eppe, Gauthier  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Mazzucchelli, Gabriel  ;  Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Language :
English
Title :
Liquid chromatography setup-dependent artefactual methionine oxidation of peptides: the importance of an adapted quality control process
Publication date :
05 August 2021
Journal title :
Journal of Chromatography. A
ISSN :
0021-9673
eISSN :
1873-3778
Publisher :
Elsevier, Amsterdam, Netherlands
Volume :
1654
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 03 September 2021

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