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2 protocols using anhydrous sodium carbonate

1

Soxhlet Extraction and Polyphenol Determination

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Petroleum ether (Carlo Erba, Spain) boiling at 40–70 °C was used for Soxhlet extraction. Chemicals for the determination of polyphenol and antioxidant content were 97% ethanol (Reanal, Hungary), 99% methanol (Reanal, Hungary), anhydrous sodium carbonate (Riedel–de Haen, Germany), Folin–Ciocalteu reagent (Merck, Germany), 2-4-6-tripyridyl-s-triazine (TPTZ) (Sigma-Aldrich, USA), acetic acid (Reanal, Hungary), anhydrous iron chloride (Merck, Germany), 98% Trolox (Sigma-Aldrich, USA), gallic acid (Sigma-Aldrich, USA).
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2

Reagents and Standards for Analytical Protocols

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Acetic acid (glacial) and anhydrous sodium carbonate were purchased from Riedel-de Haën (Hanover, Germany). Gallic acid monohydrate was obtained from Fluka (Sigma-Aldrich, Milan, Italy), methanol from VWR Prolabo (Fontenay-sous-Bois, France), and Folin-Ciocalteu's (FC) reagent from Labochimica (Padova, Italy). Chlorogenic acid hemihydrates, maltose, D-(+)-glucose, and D-(-)-fructose were purchased from Aldrich Chemical Company (Sigma-Aldrich); caffeic acid, p-coumaric acid, and ferulic acid from Sigma and methanol from Carlo Erba (Milan, Italy). Deionized water (18 MΩ) was prepared using an ultrapure water purification system (Arium ® pro; Sartorius, Muggiò, Italy). All reagents and standards were of analytical and high-performance liquid chromatography (HPLC) grade.
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