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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

Sigma-Aldrich
(−)-Epigallocatechin gallate, ≥95%
Sigma-Aldrich
Ethanol, tested according to Ph. Eur.
Sigma-Aldrich
(−)-Epigallocatechin gallate, ≥80% (HPLC), from green tea
Supelco
(−)-Epigallocatechin gallate, analytical standard
Epigallocatechin gallate, primary reference standard
Millipore
Bifido Selective Supplement B, suitable for microbiology
Sigma-Aldrich
Ethanol, BioUltra, for molecular biology, ≥99.8%, (absolute alcohol, without additive, A15 o1)
Supelco
Epigallocatechin gallate, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Methyl alcohol, United States Pharmacopeia (USP) Reference Standard
USP
Glacial acetic acid, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
Sigma-Aldrich
Taurine, BioUltra, ≥99.5% (T)
Sigma-Aldrich
Myricetin, ≥96.0% (HPLC)
Sigma-Aldrich
Ethanol, for residue analysis
Sigma-Aldrich
5α-Androstan-17β-ol-3-one, ≥97.5%
Sigma-Aldrich
Luteolin, ≥98% (TLC), powder
Sigma-Aldrich
Potassium iodide, suitable for plant cell culture
SAFC
Taurine
Sigma-Aldrich
Taurine, suitable for cell culture, meets USP testing specifications
Sigma-Aldrich
Taurine, ≥99%
Supelco
Hesperetin, analytical standard
Sigma-Aldrich
Myricetin, ≥96.0%, crystalline
Sigma-Aldrich
Gallic acid, 97.5-102.5% (titration)
Supelco
5α-Androstan-17β-ol-3-one, VETRANAL®, analytical standard
Sigma-Aldrich
Poly(tetrafluoroethylene), powder (free-flowing), ≤12 μm particle size
Sigma-Aldrich
Chrysin, ≥96.5%
Sigma-Aldrich
Poly(tetrafluoroethylene), powder (free-flowing), 1 μm particle size
Supelco
Acetic acid, analytical standard
Sigma-Aldrich
Ethanol, purum, secunda spirit, denaturated with 2% 2-butanone, S15, ~96% (based on denaturant-free substance)
Sigma-Aldrich
Acetic acid, for luminescence, BioUltra, ≥99.5% (GC)