Fawzy A. Elbarbry, Bethany Hecker, Michael J. Espiritu, Ahmed E. Elsawi, Wagdy M. Eldehna
{"title":"LC-MS /MS法同时定量人血浆中双靶向1,5-二芳基-1,2,4-三唑磺胺(WA11-13","authors":"Fawzy A. Elbarbry, Bethany Hecker, Michael J. Espiritu, Ahmed E. Elsawi, Wagdy M. Eldehna","doi":"10.1002/bmc.70165","DOIUrl":null,"url":null,"abstract":"<div>\n \n <p>We recently designed and synthesized novel dual-targeting anticancer 1,5-diaryl-1,2,4-triazole-tethered sulfonamides. Among them, WA11–13 showed promising carbonic anhydrase and VEGFR-2 inhibitory activity. This study presents a validated, sensitive LC–MS/MS assay for quantifying these compounds in human plasma. Such assay would be very important for future preclinical studies and therapeutic drug monitoring. After protein precipitation with acetonitrile, analytes and the internal standard (carbamazepine) were separated on a Phenomenex Kinetex C18 column using binary gradient elution. The mobile phase was 0.1% formic acid in water and acetonitrile (95:5, v/v) at 0.7 mL/min. Total run time was under 6 min. Detection used an API 3500 triple quadrupole mass spectrometer with electrospray ionization in positive mode. Quantification relied on multiple reaction monitoring for high sensitivity and specificity. The method was fully validated per FDA guidelines, showing acceptable linearity, accuracy, precision, selectivity, and stability. Linearity was observed in the ranges of 2.5–750 ng/mL for WA11, 25–1000 ng/mL for WA12, and 50–1000 ng/mL for WA13. The method was successfully applied to spiked human plasma, supporting its potential for therapeutic drug monitoring. Validation confirms the assay's high sensitivity, accuracy, and precision, making it suitable for future preclinical and clinical investigations of these investigational anticancer agents.</p>\n </div>","PeriodicalId":8861,"journal":{"name":"Biomedical Chromatography","volume":"39 8","pages":""},"PeriodicalIF":1.7000,"publicationDate":"2025-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"LC–MS/MS Assay for Simultaneous Quantification of Dual-Targeting 1,5-Diaryl-1,2,4-Triazole Sulfonamides (WA11–13) in Human Plasma\",\"authors\":\"Fawzy A. Elbarbry, Bethany Hecker, Michael J. Espiritu, Ahmed E. Elsawi, Wagdy M. Eldehna\",\"doi\":\"10.1002/bmc.70165\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n <p>We recently designed and synthesized novel dual-targeting anticancer 1,5-diaryl-1,2,4-triazole-tethered sulfonamides. Among them, WA11–13 showed promising carbonic anhydrase and VEGFR-2 inhibitory activity. This study presents a validated, sensitive LC–MS/MS assay for quantifying these compounds in human plasma. Such assay would be very important for future preclinical studies and therapeutic drug monitoring. After protein precipitation with acetonitrile, analytes and the internal standard (carbamazepine) were separated on a Phenomenex Kinetex C18 column using binary gradient elution. The mobile phase was 0.1% formic acid in water and acetonitrile (95:5, v/v) at 0.7 mL/min. Total run time was under 6 min. Detection used an API 3500 triple quadrupole mass spectrometer with electrospray ionization in positive mode. Quantification relied on multiple reaction monitoring for high sensitivity and specificity. The method was fully validated per FDA guidelines, showing acceptable linearity, accuracy, precision, selectivity, and stability. Linearity was observed in the ranges of 2.5–750 ng/mL for WA11, 25–1000 ng/mL for WA12, and 50–1000 ng/mL for WA13. The method was successfully applied to spiked human plasma, supporting its potential for therapeutic drug monitoring. Validation confirms the assay's high sensitivity, accuracy, and precision, making it suitable for future preclinical and clinical investigations of these investigational anticancer agents.</p>\\n </div>\",\"PeriodicalId\":8861,\"journal\":{\"name\":\"Biomedical Chromatography\",\"volume\":\"39 8\",\"pages\":\"\"},\"PeriodicalIF\":1.7000,\"publicationDate\":\"2025-07-06\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biomedical Chromatography\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/bmc.70165\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biomedical Chromatography","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/bmc.70165","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
LC–MS/MS Assay for Simultaneous Quantification of Dual-Targeting 1,5-Diaryl-1,2,4-Triazole Sulfonamides (WA11–13) in Human Plasma
We recently designed and synthesized novel dual-targeting anticancer 1,5-diaryl-1,2,4-triazole-tethered sulfonamides. Among them, WA11–13 showed promising carbonic anhydrase and VEGFR-2 inhibitory activity. This study presents a validated, sensitive LC–MS/MS assay for quantifying these compounds in human plasma. Such assay would be very important for future preclinical studies and therapeutic drug monitoring. After protein precipitation with acetonitrile, analytes and the internal standard (carbamazepine) were separated on a Phenomenex Kinetex C18 column using binary gradient elution. The mobile phase was 0.1% formic acid in water and acetonitrile (95:5, v/v) at 0.7 mL/min. Total run time was under 6 min. Detection used an API 3500 triple quadrupole mass spectrometer with electrospray ionization in positive mode. Quantification relied on multiple reaction monitoring for high sensitivity and specificity. The method was fully validated per FDA guidelines, showing acceptable linearity, accuracy, precision, selectivity, and stability. Linearity was observed in the ranges of 2.5–750 ng/mL for WA11, 25–1000 ng/mL for WA12, and 50–1000 ng/mL for WA13. The method was successfully applied to spiked human plasma, supporting its potential for therapeutic drug monitoring. Validation confirms the assay's high sensitivity, accuracy, and precision, making it suitable for future preclinical and clinical investigations of these investigational anticancer agents.
期刊介绍:
Biomedical Chromatography is devoted to the publication of original papers on the applications of chromatography and allied techniques in the biological and medical sciences. Research papers and review articles cover the methods and techniques relevant to the separation, identification and determination of substances in biochemistry, biotechnology, molecular biology, cell biology, clinical chemistry, pharmacology and related disciplines. These include the analysis of body fluids, cells and tissues, purification of biologically important compounds, pharmaco-kinetics and sequencing methods using HPLC, GC, HPLC-MS, TLC, paper chromatography, affinity chromatography, gel filtration, electrophoresis and related techniques.