Bio-guided Fractionation of Centaurea bruguierana subsp. belangeriana Extract Based on Anti-Helicobacter pylori Activity

Abstract

<strong>Background and objectives:</strong> <em>Centaurea</em> <em>bruguierana</em> subsp. <em>belangeriana</em> (DC.) Bornm. is an annual herb widely distributed in the world. It is used in folk medicine of Iran as an anti-ulcer for gastrointestinal problems. Previous studies have revealed the anti-ulcer effects of the plant extract and in the present study the anti-<em>Helicobacter</em> <em>pylori</em> activity of different fractions, isolated from <em>C. bruguierana</em> subsp. <em>belangeriana</em> was evaluated. Urease and motility inhibition activity were also examined for determination of possible mechanisms. <strong>Method:</strong> Ethanol 80% was used for the extraction. Chloroform, ethyl acetate, and methanol fractions of the total extract were obtained by solid-liquid extraction. All extracts were evaluated against jack bean urease, bacterial swarming and Minimal Inhibitory Concentrations (MICs) were determined using the agar diffusion method. Bio-guided fractionation was performed by isolation and purification of compounds from active fractions using silica-gel open column chromatography by column and thin layer chromatographic methods and identification by spectroscopic data. <strong>Results:</strong> Total extract and chloroform fraction possessed the highest anti-<em>H. pylori</em> activity with MIC 325 300 µg/mL and, respectively. The total extract was the most potent urease inhibitor (IC₅₀ 250 µg/mL). The motility test confirmed the results by inhibiting swarming at concentrations comparable with IC₅₀. The separation and purification of effective compounds of the chloroform extract was performed according to the results; Lupeol (1), Retusin (2), and Apigenin (3) were isolated and identified. <strong>Conclusion:</strong> Our research demonstrated a noticeable anti-<em>H.pylori</em> activity of different fractions from<em>C.bruguierana</em> subsp. <em>belangeriana</em> especially the nonpolar fraction.<br /> <br /> <strong> </strong>