Biomedical Chromatography 2014-07-01

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.

Cleverson Antonio Ferreira Martins, Almeriane Maria Weffort-Santos, João Cleverson Gasparetto, Angela Cristina Leal Badaró Trindade, Michel Fleith Otuki, Roberto Pontarolo

Index: Biomed. Chromatogr. 28(7) , 986-93, (2014)

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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.Copyright © 2014 John Wiley & Sons, Ltd.