Determining cyclooxygenase-2 activity in three different test systems utilizing online-solid phase extraction-liquid chromatography-mass spectrometry for parallel quantification of prostaglandin E(2), D(2) and thromboxane B(2)

Journal of Chromatography. a
Ina WillenbergNils Helge Schebb

Abstract

Cyclooxygenase-2 (COX-2) catalyzes the formation of PGH2 from arachidonic acid. PGH2 is further converted to different prostaglandins (PG), such as PGE2, PGD2 and TxB2. In this study a rapid online-SPE-LC-MS method for the simultaneous quantification of PGE2, PGD2 and TxB2 streamlined for COX-2 enzyme assays is presented. Baseline separation of all analytes was achieved in only 7.1 min per sample, including sample preparation by online SPE. The method showed high sensitivity (LODs of 0.65-1.25 fmol on column) and accuracy (89-113%) in protein containing media. Because of online-SPE, no manual sample preparation was required, except for addition of IS solution, allowing to use the approach as rapid read-out in COX-2 activity assays. This was demonstrated by applying the method on three in vitro test systems: a cell-free enzyme assay, an assay using HCA-7 cells constitutively expressing COX-2 and primary human monocytes. In these assays, the potency of three popular drugs celecoxib, indomethacin and dexamethasone was successfully characterized with the new online-LC-MS method. The comparison of the results showed that the inhibitory effects of PG formation strongly depend on the test system. Thus we suggest that the modulation of...Continue Reading

References

Jul 26, 1984·The New England Journal of Medicine·C C HardyS T Holgate
Aug 10, 2000·The Journal of Biological Chemistry·J ShaoR N DuBois
Nov 4, 2000·Biochemical and Biophysical Research Communications·C M ReddyK M Madyastha
Sep 21, 2002·Proceedings of the National Academy of Sciences of the United States of America·N V ChandrasekharanDaniel L Simmons
Nov 16, 2002·Prostaglandins & Other Lipid Mediators·Ikuo Morita
Aug 20, 2004·Pharmacological Reviews·Daniel L SimmonsTimothy Hla
Jul 4, 2006·Bioorganic & Medicinal Chemistry Letters·Wei-Jern TsaiLi-Ming Yang
Dec 1, 2006·The Journal of Biological Chemistry·Richard HarkewiczEdward A Dennis
May 10, 2007·Nature Protocols·Muriel CuendetJohn M Pezzuto
Sep 20, 2007·Pharmacological Reviews·Bengt SamuelssonPer-Johan Jakobsson
Feb 27, 2009·Journal of Lipid Research·Matthew W BuczynskiEdward A Dennis
Sep 8, 2010·Journal of Pharmaceutical and Biomedical Analysis·Hongmei CaoRichard B van Breemen
Apr 22, 2011·Arteriosclerosis, Thrombosis, and Vascular Biology·Emanuela Ricciotti, Garret A FitzGerald
Jun 22, 2011·Biochimica Et Biophysica Acta·Darren S DumlaoEdward A Dennis
Aug 26, 2011·The Journal of Pharmacology and Experimental Therapeutics·Rui YuRichard B van Breemen
Oct 4, 2011·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Ayako FurugenKen Iseki
Apr 17, 2013·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Linda KortzUta Ceglarek
Jan 13, 2015·Analytical and Bioanalytical Chemistry·Ina WillenbergNils Helge Schebb

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Citations

Apr 22, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Anna Catharina SuhrMichael Vogeser
Jun 24, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Chang ShuWansheng Chen
Jan 24, 2017·PloS One·Shan WangMichael Telamon Empl
Nov 27, 2017·Metabolomics : Official Journal of the Metabolomic Society·Junjie ZhangChun Cai
Aug 18, 2020·American Journal of Physiology. Renal Physiology·Robert GreiteFaikah Gueler
Apr 6, 2021·Frontiers in Cellular and Infection Microbiology·Yingxin SunQing Gao

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