We used the same technical setup as for calibration (Supplementary Materials, File 1 ). A total of 30 µL of an animal-sourced mixture of polyunsaturated fatty acids and three isolated polyunsaturated fatty acids—linoleic, linolenic, and arachidonic acid (analytical standard, Merck, Darmstadt, Germany)—were oxidized in a cleaned flask under 100 mL/min flow of highly purified synthetic air (oxygen content: 21%; Alphagaz 1, Air Liquide, Paris, France) and constant fanning. Headspace gas was sampled at 10-min intervals by the MCC–IMS. Signal intensities between the limit of detection and quantification were considered as unquantifiable traces.
Linolenic
Manufactured by Merck Group
Sourced in Germany
Linolenic is a laboratory equipment product manufactured by Merck Group. It is designed to perform a specific function in the laboratory setting, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation or interpretation.
Automatically generated - may contain errors
Lab products found in correlation
Stearic, by Merck Group (4 mentions)
Linoleic acid, by Merck Group (2 mentions)
Arachidonic acid, by Merck Group (1 mentions)
Alphagaz 1, by Air Liquide (1 mentions)
Millipore q system, by Merck Group (1 mentions)
Cholesterol, by Merck Group (1 mentions)
Campesterol, by Merck Group (1 mentions)
Stigmasterol, by Merck Group (1 mentions)
Brassicasterol, by Merck Group (1 mentions)
Toluene, by Merck Group (1 mentions)
Suberic, by Merck Group (1 mentions)
Sebacic, by Merck Group (1 mentions)
Lsm 510, by Zeiss (1 mentions)
Nile red oxazone, by Merck Group (1 mentions)
Hplc grade solvent, by Merck Group (1 mentions)
Palmitoleic, by Merck Group (1 mentions)
Arachidic, by Merck Group (1 mentions)
Oleacein, by Phytolab (1 mentions)
Oleocanthal, by Phytolab (1 mentions)
Myristic, by Merck Group (1 mentions)
7 protocols using linolenic
1
Oxidation of Polyunsaturated Fatty Acids
2
Fatty Acid and Sterol Analysis Protocol
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A Millipore-Q system (Millipore Corp., Saint Quentin, France) was used to purify water. The standards of fatty acids (oleic, palmitic, palmitoleic, myristic, margaric, stearic, linoleic, arashidic, margoleic, gadoleic, linolenic, lignoceric and behenic acids) were obtained from Merck (Darmstadt, Germany) . All chemicals and standards (cholesterol, brassicasterol, 24-methylen cholesterol, campesterol, campestanol, stigmasterol, Δ7-campesterol, Δ5,23stigmastadienol, clerosterol, b-sistosterol, sitostanol, Δ5avenasterol, Δ5,24-stigmastadienol, Δ7-stigmastenol, Δ7avenasterol, erythrodiol and uvaol) used in sterol analysis were also purchased from Merck except for internal standards (3-α-cholestanol) from Sigma-Aldrich, Supelco (Bellefonte, PA, USA) . All chemicals and solvents used in the analyzes were of GC and HPLC purity (99.0％) .
3
Fatty Acid Analysis by GC-MS
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Methanol and n-hexane of analytical grade were obtained from PENTA (Praha, Czech Republic). The toluene (analytical-grade) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Further, the 3-(Trifluoromethyl)phenyltrimethylammonium hydroxide (TFTMAH), 5% w/v in methanol (known as MethPrep II), was supplied by Tokyo Chemical Industry Co., Ltd. (Nihonbashi-honcho, Chuo-ku, Tokyo, Japan). The fatty acid standards—azelaic (C9), suberic (C8), sebacic (C10), palmitic (C16:0), stearic (C18:0), oleic (C18:1), linoleic (C18:2), linolenic (C18:3), and cholesta-3,5-diene-7-one—were purchased from Sigma-Aldrich. Individual stock standard solutions were prepared for fatty acids in n-hexane and cholesta-3,5-diene-7-one in 2-propanol at a concentration of 1 mg·mL−1 and stored in a refrigerator at 5 °C.
4
Lipid Droplet Visualization in Fungi
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Mycelia were incubated in MM containing 80 mg l−1 toxoflavin and 1% sodium oleate, 1% linoleic acid, or 1% linolenic (Sigma-Aldrich) at 25 °C with shaking at 200 r.p.m. for 24 h. Lipid droplets in fungal cells were stained with a Nile Red solution consisting of 50 mM Tris/maleate buffer (pH 7.5), 20 mg ml−1 polyvinylpyrrolidone, and 2.5 μg ml−1 Nile Red Oxazone (Sigma-Aldrich). The samples were incubated for 15 min at room temperature and were washed two times with phosphate-buffered saline. Fluorescence emitted by the lipid droplets was observed with a Carl Zeiss confocal microscope (excitation at 490 nm, LSM 510). To determine whether vegetative growth that was affected by toxoflavin could be complemented with exogenous sodium oleate, 1% sodium oleate was added into MM agar supplemented with 80 mg l−1 toxoflavin or 5 mM H2O2. Each fungal strain was incubated at 25 °C for 4 d.
5
Quantitative Analysis of Olive Oil Compounds
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High purity standards for the qualitative-quantitative determination of fatty acids (palmitic, palmitoleic, heptadecanoic, stearic, oleic, linoleic, linolenic, arachidic, eicosenoic, behenic), α-T, triterpenic acids (maslinic and oleanoic acids), hexanal, and 2-methylpropyl acetate, as well as all High-Performance Liquid Chromatography (HPLC) grade solvents, were all purchased from Sigma–Aldrich (Milan, Italy). Tyrosol, hydroxytyrosol, and verbascoside were purchased from Fluka (Milan, Italy), Cabru s.a.s. (Arcore, Milan, Italy), and Extrasynthese (Genay Cedex, France), respectively. Oleacein and oleocanthal were obtained from PhytoLab GmbH & Co. (Vestenbergsgreuth, Germany). Carotenoids (lutein and β-carotene) and β-T3 standards were purchased from CaroteNature (Lupsingen, Liestal, Switzerland) and Cayman chemicals (Ann Arbor, MI, USA), respectively.
6
Fatty Acid Profiling of Oils
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Fatty acids of oils were converted into their methyl esters (FAMEs) before gas chromatography (GC) analyses with a modification according to the AOCS Official Method Ce 1h-05 (Firestone, 2009 ) were performed. Oils were dissolved in 0.5 mL hexane and converted into FAMEs and 100 μL methanolic KOH (2 M) was added. Hydrochloric acid (2 M) was added until methyl orange indicator changed to pink and the mixture was allowed to settle. Then, 10 μL of the organic layer was injected into an Agilent 7890 GC (Agilent Technologies, Santa Clara, CA) equipped with a flame ionization detector. A stainless-steel column (30 m × 0.25 mm) packed with 70% cyanopropyl polysilphenylene siloxane was used. The oven temperature was held at 100 °C after sample injection and increased to 225 °C with a rate of 5 °C/min. The injector and detector temperatures were 260 °C and 280 °C, respectively. Helium was used as the carrier gas (3 mL/min), split ratio of 1:100 and the injection volume was 1 μL. The concentration of FAMEs in samples was determined using fatty acids standards including myristic, palmitic, stearic, oleic, linoleic, linolenic and arachidic acid (Sigma Aldrich, UK).
7
Effects of Fatty Acids on Meibomian Gland Cells
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