A Robust LC-MS/MS Method for Determination of Schiff-base Forming Reversible Covalent Inhibitors in Whole Blood

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February 10,2026

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Covalent inhibitors are small molecules that bind to target proteins via covalent bonds, enhancing inhibitory activity. They are classified as reversible or irreversible inhibitors based on bond type. Voxelotor, a sickle cell disease inhibitor, uses its aldehyde group to reversibly form a Schiff base with Val1 amino group on hemoglobin's beta chain. This stabilizes hemoglobin's R-state via intramolecular hydrogen bonds, boosting oxygen affinity. Optimized pretreatment overcomes extremely low extraction recovery of Schiff-Base forming reversible covalent compounds in whole blood.

A-Robust-LC-MSMS-Method-for-Determination-of-Schiff-base-Forming-Reversible-Covalent-Inhibitors-in-Whole-Blood.jpg

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