Development and Validation of LC–MS/MS with In-Source Collision-Induced Dissociation for The Quantification of Pegcantratinib in Human Skin Tumors

Abstract

Aim: Pegcantratinib is a mini-PEGylated K252a derivative, under clinical evaluation as an anticancer agent acting through inhibition of the tropomyosin receptor kinase. A method for quantifying pegcantratinib in skin tumor biopsies of patients was required to determine tumor drug penetration. Methods & results: A sensitive and PEGylated molecule specific HPLC-MS/MS method coupled with in-source collision-induced dissociation was developed. The method exhibited excellent precision (coefficient of variation ≤8.5%), accuracy in the range 95-102%, high and consistent recovery and no matrix effect. The assay was linear across a range of 1-500 ng/ml, with a limit of quantitation of 2.5 ng/ml. Conclusion: We have developed and validated a method for analyzing pegcantratinib in human tumor biopsies, with the approach successfully applied to clinical trial samples.

In‐source collision‐induced dissociation (IS‐CID): Applications, issues and structure elucidation with single‐stage mass analyzers

“…An alternative approach has been developed for the analysis of PEGylated peptides. In this case, IS‐CID was used to release the unconjugated peptide or therapeutic agent moiety that could subsequently be analyzed by MRM of the collision fragments produced in the ESI source . The peptide fragments can be selected and analyzed as surrogates for the PEGylated peptides.…”