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Phytochemical analysis and in vitro biological activity of three Hypericum species from the Canary Islands (Hypericum reflexum, Hypericum canariense and Hypericum grandifolium).

Fitoterapia (2014-12-03)
Christian Zorzetto, Candelaria C Sánchez-Mateo, Rosa M Rabanal, Giulio Lupidi, Dezemona Petrelli, Luca A Vitali, Massimo Bramucci, Luana Quassinti, Giovanni Caprioli, Fabrizio Papa, Massimo Ricciutelli, Gianni Sagratini, Sauro Vittori, Filippo Maggi
ABSTRACT

In the present work we carried out a phytochemical and biological investigation on three Hypericum species, i.e. Hypericum reflexum, Hypericum canariense and Hypericum grandifolium, from the Canary Islands where they are traditionally used as diuretic, wound healing, vermifuge, sedative and antidepressive agents. The polar extracts of the top flowering aerial parts, prepared by Soxhlet apparatus using a methanol-acetone (1:1) extracting mixture, were analyzed by HPLC-DAD and HPLC-MS for the content of eight biomarkers such as hypericin, hyperforin, chlorogenic acid, rutin, hyperoside, isoquercitrin, quercitrin and quercetin, whereas the hydrodistilled essential oils were analyzed by GC-FID and GC-MS. The three Hypericum species had different results in both polar and volatile constituents, H. reflexum being the only one endowed with a small amount of naphtodianthrones (hypericin and pseudohypericin), and containing high levels of chlorogenic acid, rutin and volatile mono- and sesquiterpenes. After chemical characterization, all products were in vitro biologically assayed for antiproliferative activity on human tumor cell lines by MTT assay, for antioxidant potential by DPPH, ABTS and FRAP assays, and for antimicrobial activity by the agar disc diffusion and microdilution methods. Results revealed interesting bioactivities and differences between polar extracts and essential oils, with the former being endowed with significant antioxidant activity and the latter with comparable inhibition effects on the tumor cells (A375, MDA-MB 231 and HCT 116) to that of cisplatin.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Methanol, suitable for HPLC, ≥99.9%
Sigma-Aldrich
Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
Sigma-Aldrich
Methanol, anhydrous, 99.8%
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Methanol, HPLC Plus, ≥99.9%
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Methanol, HPLC Plus, ≥99.9%, poly-coated bottles
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Methanol, suitable for HPLC, gradient grade, ≥99.9%
Sigma-Aldrich
Methanol, suitable for HPLC, gradient grade, suitable as ACS-grade LC reagent, ≥99.9%
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Acetone, ACS reagent, ≥99.5%
Sigma-Aldrich
Acetonitrile, anhydrous, 99.8%
Supelco
Methanol, analytical standard
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Ethyl acetate, anhydrous, 99.8%
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Acetone, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.9%
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Hexane, anhydrous, 95%
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Methanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)
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Acetonitrile, HPLC Plus, ≥99.9%
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Ethyl acetate, ACS reagent, ≥99.5%
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Methanol, ACS reagent, ≥99.8%
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Quercetin, ≥95% (HPLC), solid
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Acetone, suitable for HPLC, ≥99.9%
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Phosphoric acid, 85 wt. % in H2O, 99.99% trace metals basis
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Ethyl acetate, suitable for HPLC, ≥99.7%
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Hexane, suitable for HPLC, ≥97.0% (GC)
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Palmitic acid, ≥99%
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Hexane, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99% (GC)
Sigma-Aldrich
Ethyl acetate, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5% (GC)
Sigma-Aldrich
Phosphoric acid, ACS reagent, ≥85 wt. % in H2O
Sigma-Aldrich
Acetone, puriss., meets analytical specification of Ph. Eur., BP, NF, ≥99% (GC)
Sigma-Aldrich
Acetonitrile, for HPLC, for UV, ≥99.9% (GC)
Sigma-Aldrich
Hexane, suitable for HPLC, ≥95%
Sigma-Aldrich
Dodecane, anhydrous, ≥99%

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