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  • Optimization of microwave-assisted extraction of polyphenols from herbal teas and evaluation of their in vitro hypochlorous acid scavenging activity.

Optimization of microwave-assisted extraction of polyphenols from herbal teas and evaluation of their in vitro hypochlorous acid scavenging activity.

Journal of agricultural and food chemistry (2014-11-05)
Burcu Bekdeşer, Nazan Durusoy, Mustafa Özyürek, Kubilay Güçlü, Reşat Apak
ABSTRACT

Hypochlorous acid (HOCl) is an important reactive oxygen species (ROS) and non-radical and is taking part in physiological processes concerned with the defense of the organism, but there has been limited information regarding its scavenging by polyphenols. This study was designed to examine the HOCl scavenging activity of several polyphenols and microwave-assisted extracts of herbal teas. HOCl scavenging activity has usually been determined spectrophotometrically by a KI/taurine assay at 350 nm. Because some polyphenols (i.e., apigenin and chrysin) have a strong ultraviolet (UV) absorption in this range, their HOCl scavenging activity was alternatively determined without interference using resorcinol (1,3-dihydroxybenzene) as a fluorogenic probe. In the present assay, HOCl induces the chlorination of resorcinol into its non-fluorescent products. Polyphenols as HOCl scavengers inhibit the chlorination of the probe by this species. Thus, the 25% inhibitive concentration (IC25) value of polyphenols was determined using the relative increase in fluorescence intensity of the resorcinol probe. The HOCl scavenging activities of the test compounds decreased in the order: epigallocatechin gallate > quercetin > gallic acid > rutin > catechin > kaempferol. The present study revealed that epigallocatechin gallate (IC25 = 0.1 μM) was the most effective scavenging agent. In addition to polyphenols, four herbal teas were evaluated for their HOCl activity using the resorcinol method. The proposed spectrofluorometric method was practical, rapid, and less open to interferences by absorbing substances in the range of 200-420 nm. The results hint to the possibility of polyphenols having beneficial effects in diseases, such as atherosclerosis, in which HOCl plays a pathogenic role.

MATERIALS
Product Number
Brand
Product Description

Rosmarinic acid, primary reference standard
Chlorogenic acid, primary reference standard
Sigma-Aldrich
Methanol, NMR reference standard
Sigma-Aldrich
Naringenin, natural (US), 98%
Supelco
Resorcinol, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
Acetic acid-12C2, 99.9 atom % 12C
Sigma-Aldrich
Quercetin, ≥95% (HPLC), solid
USP
Dehydrated Alcohol, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Potassium iodide, ≥99.99% trace metals basis
Sigma-Aldrich
Potassium iodide, AnhydroBeads, −10 mesh, 99.998% trace metals basis
Sigma-Aldrich
Methanol, anhydrous, 99.8%
Sigma-Aldrich
Resorcinol, ACS reagent, ≥99.0%
Sigma-Aldrich
4-Chlororesorcinol, 98%
Supelco
Ethanol solution, certified reference material, 2000 μg/mL in methanol
Supelco
Acetic acid, analytical standard
Supelco
Methanol, analytical standard
Sigma-Aldrich
Potassium iodide, BioUltra, ≥99.5% (AT)
Sigma-Aldrich
Kaempferol, ≥97.0% (HPLC)
Sigma-Aldrich
Resorcinol, ≥98%, FG
Sigma-Aldrich
Acetic acid, ≥99.5%, FCC, FG
Sigma-Aldrich
Acetic acid, natural, ≥99.5%, FG
Sigma-Aldrich
Resorcinol, ReagentPlus®, 99%
Sigma-Aldrich
Rosmarinic acid, 96%
Sigma-Aldrich
Apigenin, ≥95.0% (HPLC)
Sigma-Aldrich
Potassium iodide, tested according to Ph. Eur.
Sigma-Aldrich
Kaempferol, ≥90% (HPLC), powder
Sigma-Aldrich
Potassium iodide, suitable for plant cell culture
Sigma-Aldrich
(±)-Naringenin, ≥95%
Sigma-Aldrich
Chlorogenic acid, ≥95% (titration)
Sigma-Aldrich
Resorcinol, BioXtra, ≥99%