ArticleJournal of mass spectrometry and advances in the clinical lab2025
A liquid chromatography-high-resolution mass spectrometry method for separation and identification of hemoglobin variant subunits with mass shifts less than 1 Da.
Article in Journal of mass spectrometry and advances in the clinical lab, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 1 paper.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
1 citing paper in PubMed.
- Sample preparation and cleanup methods for clinical top-down proteomics.Expert review of proteomics · 2025Review
Corrections and comments
- Erratum issued
Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Background: Identification of hemoglobin (Hb) variants is valuable in clinical testing. A common issue with conventional methods for identifying Hb variants is their subpar ability to provide structural breakdowns of the variants. Reports have surfaced of high-resolution mass spectrometry (HR-MS) methods that improve on traditional methods; however, ambiguities may arise without separation of Hb subunits prior to HR-MS analysis. Methods: We report a liquid chromatography-high-resolution mass spectrometry (LC-HR-MS) method to separate several pairs of normal and variant Hb subunits with mass shifts of less than 1 Da and successfully identify them in intact-protein and top-down analyses. LC separation was facilitated by a C4 reversed-phase column. Results: Seven heterozygous Hb variant samples (Hb C with α-thalassemia trait, Hb E, Hb D-Punjab, Hb G-Accra, Hb G-Siriraj, Hb Tarrant, and Hb G-Waimanalo) were selected to demonstrate the LC separation of Hb variant and normal subunits with mass shifts of less than 1 Da. The analytes could be explicitly observed in the deconvoluted MS Conclusions: The LC-HR-MS method described can effectively separate and identify Hb subunits, especially when the variant subunits have mass deviations of less than 1 Da from their corresponding normal subunits. With further evaluation to prove the clinical utility, the HR-MS methods including CE-HR-MS have the potential to complement or partially replace conventional methods of Hb variant identification in clinical laboratories.
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