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3 protocols using linoleic acid

1

Extraction and Isolation of Natural Compounds

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Solvents for different steps in analyses were partially denatured ethanol (AustrAlco, Spillern, Austria) for extraction, hexane, ethyl acetate (both Carl Roth, Karlsruhe, Baden-Württemberg, Germany) and methanol (VWR, Radnor, PA, USA) for open column chromatography and TLC, chloroform-d1 (VWR) and pyridine-d5 (Armar, Döttingen, Aargau, Switzerland) for NMR.
HPLC mobile phases consisted of ultrapure water prepared from deionized water with a Barnstead MicroPure system (Thermo Fisher Scientific, Waltham, MA, USA), HPLC-grade acetonitrile (VWR) and formic acid for LC-MS (Honeywell, Charlotte, NC, USA).
Reference substances were abietic acid contributed by the Department of Pharmaceutical Chemistry, University of Graz, Austria, apigenin (Sigma-Aldrich, St. Louis, MO, USA), chrysin, pinocembrin-7-methyl ether (pinostrobin), linoleic acid (all Carl Roth) and oleic acid (Fluka, Buchs, St. Gallen, Switzerland).
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2

Extraction and Isolation of Natural Compounds

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Solvents for different steps in analyses were partially denatured ethanol (AustrAlco, Spillern, Austria) for extraction, hexane, ethyl acetate (both Carl Roth, Karlsruhe, Baden-Württemberg, Germany) and methanol (VWR, Radnor, PA, USA) for open column chromatography and TLC, chloroform-d1 (VWR) and pyridine-d5 (Armar, Döttingen, Aargau, Switzerland) for NMR.
HPLC mobile phases consisted of ultrapure water prepared from deionized water with a Barnstead MicroPure system (Thermo Fisher Scientific, Waltham, MA, USA), HPLC-grade acetonitrile (VWR) and formic acid for LC-MS (Honeywell, Charlotte, NC, USA).
Reference substances were abietic acid contributed by the Department of Pharmaceutical Chemistry, University of Graz, Austria, apigenin (Sigma-Aldrich, St. Louis, MO, USA), chrysin, pinocembrin-7-methyl ether (pinostrobin), linoleic acid (all Carl Roth) and oleic acid (Fluka, Buchs, St. Gallen, Switzerland).
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3

Analysis of Fatty Acid Derivatives

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All chemicals used were standard commercial products of analytical grade from the following companies: linoleic acid (Roth, Karlsruhe, Germany); α-linolenic acid (Roth); arachidonic acid (Sigma-Aldrich, Steinheim, Germany); triphenyl phosphine (Merck Schuchardt, Hohenbrunn, Germany); HPLC solvents (Mallinckrodt Baker, Deventer, Holland); 9(±)-HODE und 13(±)-HODE (Cayman Chemical, Tallinn, Estonia); Hydroperoxy HPLC Mix (Cayman Chemical).
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