Benzoyl chloride derivatization improves selectivity and sensitivity of lipidomic quantitation in human serum of pancreatic cancer patients using RP-UHPLC/MS/MS

Chemical derivatization using benzoyl chloride significantly enhances the chromatographic behavior and mass spectrometric sensitivity of lipids, which are crucial in cellular processes and cancer biomarker research. This study presents a targeted and validated method that combines benzoyl chloride derivatization with reversed-phase ultrahigh-performance liquid chromatography tandem mass spectrometry (RP-UHPLC/MS/MS) for quantitative lipidomic analysis of human serum. A total of 450 lipid species from 19 lipid subclasses were identified using multiple reaction monitoring transitions, retention dependencies, and derivatization tags. The method improves sensitivity especially for monoacylglycerols, diacylglycerols, sphingoid bases, and free sterols, while maintaining quantitative accuracy verified with NIST SRM 1950 plasma reference material. Lipidomic profiling of pancreatic cancer patients versus healthy controls revealed significant dysregulation in lipid metabolism, characterized by upregulation of monoacylglycerols and sphingosine and downregulation of sphingolipids with very long saturated N-acyl chains and phospholipids containing 18:2 and 20:4 fatty acyl compositions. The developed approach provides new structural insights into lipid metabolic alterations in pancreatic cancer and demonstrates high potential for biomarker discovery and clinical lipidomics applications.