{"title":"Algorithm-Driven Chromatographic Method for Prostaglandin Isomer Identification via Tandem Mass Spectrometry","authors":"Toshinobu Hondo*, , , Yumi Miyake, , and , Michisato Toyoda, ","doi":"10.1021/jasms.5c00231","DOIUrl":null,"url":null,"abstract":"<p >This study explores the computational isolation of prostaglandin (PG) isomers, specifically PG E<sub>2</sub> (PGE<sub>2</sub>) and D<sub>2</sub> (PGD<sub>2</sub>), to enhance method development efficiency and provide insights into their retention behavior during supercritical fluid extraction (SFE) combined with supercritical fluid chromatography (SFC)-tandem mass spectrometry (MS/MS). Although PGE<sub>2</sub> and PGD<sub>2</sub> are positional isomers that yield identical product ions in MS/MS, they serve distinct biological roles. This research illustrates the efficacy of selected reaction monitoring (SRM)-based techniques for differentiating coeluting isomers. Despite the challenges posed by baseline resolution, simplified computational methods successfully distinguished between PGE<sub>2</sub> and PGD<sub>2</sub>, demonstrating the potential for high-throughput PG analysis without the necessity for complete chromatographic peak resolution. By employing least-squares estimation to solve a linear system, the abundance ratio of PGE<sub>2</sub> to PGD<sub>2</sub> was derived from intensity ratios across four SRM transitions, achieving precise quantification even with poorly resolved SFC peaks. The study highlights critical factors affecting PG retention, such as the choice of the stationary phase, temperature regulation, and reduction of stainless steel interactions, which can diminish signal intensity. A significant observation is the concentration-dependent suppression effect of the entrainer when interacting with the hepatocyte matrix, underscoring the importance of effective matrix management in SFE/SFC-MS/MS. These findings advance the development of a robust, high-throughput analytical platform for PG quantification and lipidomics research applications.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"36 10","pages":"2267–2275"},"PeriodicalIF":2.7000,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of the American Society for Mass Spectrometry","FirstCategoryId":"92","ListUrlMain":"https://pubs.acs.org/doi/10.1021/jasms.5c00231","RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
This study explores the computational isolation of prostaglandin (PG) isomers, specifically PG E2 (PGE2) and D2 (PGD2), to enhance method development efficiency and provide insights into their retention behavior during supercritical fluid extraction (SFE) combined with supercritical fluid chromatography (SFC)-tandem mass spectrometry (MS/MS). Although PGE2 and PGD2 are positional isomers that yield identical product ions in MS/MS, they serve distinct biological roles. This research illustrates the efficacy of selected reaction monitoring (SRM)-based techniques for differentiating coeluting isomers. Despite the challenges posed by baseline resolution, simplified computational methods successfully distinguished between PGE2 and PGD2, demonstrating the potential for high-throughput PG analysis without the necessity for complete chromatographic peak resolution. By employing least-squares estimation to solve a linear system, the abundance ratio of PGE2 to PGD2 was derived from intensity ratios across four SRM transitions, achieving precise quantification even with poorly resolved SFC peaks. The study highlights critical factors affecting PG retention, such as the choice of the stationary phase, temperature regulation, and reduction of stainless steel interactions, which can diminish signal intensity. A significant observation is the concentration-dependent suppression effect of the entrainer when interacting with the hepatocyte matrix, underscoring the importance of effective matrix management in SFE/SFC-MS/MS. These findings advance the development of a robust, high-throughput analytical platform for PG quantification and lipidomics research applications.
期刊介绍:
The Journal of the American Society for Mass Spectrometry presents research papers covering all aspects of mass spectrometry, incorporating coverage of fields of scientific inquiry in which mass spectrometry can play a role.
Comprehensive in scope, the journal publishes papers on both fundamentals and applications of mass spectrometry. Fundamental subjects include instrumentation principles, design, and demonstration, structures and chemical properties of gas-phase ions, studies of thermodynamic properties, ion spectroscopy, chemical kinetics, mechanisms of ionization, theories of ion fragmentation, cluster ions, and potential energy surfaces. In addition to full papers, the journal offers Communications, Application Notes, and Accounts and Perspectives