title: Malva sylvestris L. extract suppresses desferrioxamine-induced PGE₂ and PGD₂ release in differentiated U937 cells: the development and validation of an LC-MS/MS method for prostaglandin quantification.
abstract: Malva sylvestris is a species used worldwide as an alternative to anti-inflammatory therapies; however, its mechanism of action remains unknown. In this paper, the anti-inflammatory effects of M. sylvestris alcoholic extracts were evaluated by measuring the pro-inflammatory mediators PGE₂ and PGD₂ in desferrioxamine-stimulated phorbol 12-myristate 13-acetate-differentiated U937 cells. An HPLC-DAD fingerprint of the M. sylvestris extract was performed and caffeic acid, ferulic acid and scopoletin were identified and quantified. An HPLC-MS/MS method was developed and validated to separate and measure the prostaglandins. The lower limits of detection (~0.5 ng/mL for PGE₂ and PGD₂) and quantification (1.0 ng/mL for PGE₂ and PGD₂) indicated that the method is highly sensitive. The calibration curves showed excellent coefficients of correlation (r > 0.99) over the range of 1.0-500.0 ng/mL, and at different levels, the accuracy ranged from 96.4 to 106.4% with an RSD < 10.0% for the precision study. This method was successfully applied using U937-d cells. A significant dose-dependent reduction of PGE2 and PGD2 levels occurred using 10 µg/mL (10.74 ± 2.86 and 9.60 ± 6.89%) and 50 µg/mL of extract (48.37 ± 3.24 and 53.06 ± 6.15%), suggesting that the anti-inflammatory mechanisms evoked by M. sylvestris may be related to modulation of these mediators.
authors.affiliation: Laboratory of Biopharmacy, Department of Pharmacy, Universidade Federal do Paraná, Brazil.
authors.forename: Cleverson Antonio Ferreira
authors.initials: CA
authors.lastname: Martins
authors.forename: Almeriane Maria
authors.initials: AM
authors.lastname: Weffort-Santos
authors.forename: João Cleverson
authors.initials: JC
authors.lastname: Gasparetto
authors.forename: Angela Cristina Leal Badaró
authors.initials: AC
authors.lastname: Trindade
authors.forename: Michel Fleith
authors.initials: MF
authors.lastname: Otuki
authors.forename: Roberto
authors.initials: R
authors.lastname: Pontarolo
mesh_terms: D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D002853:Chromatography, Liquid / Q000379:methods; D003676:Deferoxamine / Q000494:pharmacology; D015232:Dinoprostone / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D028501:Malva; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D015230:Prostaglandin D2 / Q000032:analysis* / Q000378:metabolism; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods; D020298:U937 Cells
chemical_list: D000893:Anti-Inflammatory Agents; D010936:Plant Extracts; D003676:Deferoxamine; D015232:Dinoprostone; D015230:Prostaglandin D2
keywords: LC-MS/MS; Malva sylvestris; PGD2; PGE2; anti-inflammatory activity