Dioleoylglycerol

N. gaditana dry biomass was obtained from AlgaEnergy (Madrid, Spain). The N. gaditana extract was obtained and analyzed in terms of lipid classes and fatty acid profile following previously optimized methods [1 (link)]. SOFTISAN^® 649 (bis-diglyceryl Polyacyladipate-2), a lanolin substitute widely use in the industry [43 (link)], was obtained from Sasol Germany GmbH & Co (Witten, Germany). Tween^® 80 (polysorbate 80) was purchased from Sigma Aldrich (Madrid, Spain). Soy lecithin was purchased from Acopharma (Barcelona, Spain). Ethyl acetate was from Vaz Pereira S.A. (Sintra, Portugal). MilliQ water was obtained from a MilliQ plus system (Millipore, Germany). Methanol was purchased from Lab Scan Analytical Sciences (Gliwice, Poland). The HPLC-grade solvents (2,2,4-trimethyl pentane, methyl tert-butyl ether (MTBE)) were purchased from Macron Fine Chemicals (Gliwice, Poland). Absolute ethanol (PRS grade), sodium hydrogen carbonate, and potassium hydroxide were purchased from Panreac Química S.A (Barcelona, Spain). Glyceryl trilinoleate, dioleoylglycerol (mixture of 1,3- and 1,2-isomers), 1-oleoyl-rac-glycerol, phosphatidylethanolamine, mono galactosyl glycerol, di galactosyl glycerol, oleic acid, and ethyl linoleate used as HPLC standards were purchased from Sigma-Aldrich (St. Louis, MO, USA). All other reagents and solvents used were of analytical or HPLC grade.

Standards for gas chromatography (oleic acid, ≥ 99%), high performance liquid chromatography (nonadecanoic acid methyl ester, 99%), and thin layer chromatography (glyceryl trioleate, ≥ 99%; dioleoylglycerol, ≥ 99%; 1-oleoyl-rac-glycerol, ≥ 99%; and oleic acid, ≥ 99%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). For gas chromatography, derivatization of all fatty acids (both esterified and free fatty acids) was achieved using acetyl chloride (≥ 99%, Sigma-Aldrich (St. Louis, MO, USA)), while derivitiation of free fatty acids alone was accomplished using diazomethane prepared from diazald (TLC Pharmaceutical Standards Ltd., Aurora, ON, Canada) and a Diazald® kit from Sigma-Aldrich (St. Louis, MO, USA). For thin layer chromatography, iodine (≥ 99.99%) and diethyl ether (≥ 98%) were also purchased from Sigma-Aldrich (St. Louis, MO, USA). The solvents used for analysis, methanol (HPLC grade, > 99.9%), toluene (HPLC grade, > 99.9%), hexane (HPLC grade, > 99.9%), acetic acid (> 99.85%), along with the o-phosphoric acid (85%) used for acidification, were all obtained from Fisher Scientific (Fairlawn, NJ, USA). Nitrogen gas (99.998%) was obtained from Praxair (Mississauga, ON, Canada).

Commercial dry powder biomass from microalga Isochrysis galbana was purchased from Neoalgae Micro Seaweed Products (Gijón, Spain). Chloroform and isopropyl alcohol were purchased from Scharlab S.L. (Sentmenat, Spain) Methanol was purchased from Lab Scan Analytical Sciences (Gliwice, Poland). Hexane and the solvents (2,2,4-trimethyl pentane, methyl tert-butyl ether (MTBE)) used for HPLC analyses were HPLC-grade and purchased from Macron Fine Chemicals (Gliwice, Poland). Absolute ethanol (PRS grade), sodium hydrogen carbonate and potassium hydroxide were purchased from Panreac Quimica S.A (Barcelona, Spain). The water used was Milli-Q grade (Millipore Sigma, Burlington, MA , USA). Viscozyme^®® from Aspergillus aculeatus containing a wide range of carbohydrases, including arabinase, cellulase, beta-glucanase, hemicellulase and xylanase, and Celluclast^®® containing cellulase from Trichoderma reesei were kindly donated by Novozymes (Bagsvaerd, Denmark). Fatty acid methyl esters standard (Supelco 37 FAME Mix) was from Supelco (Bellefonte, PA, USA). Glyceryl trilinoleate, dioleoylglycerol (mixture of 1,3- and 1,2-isomers), 1-oleoyl-rac-glycerol, oleic acid and ethyl linoleate used as HPLC standards was purchased from Sigma-Aldrich (St. Louis, MO, USA). All other reagents and solvents used were of analytical or HPLC grade.

2,4-D, butylated hydroxytoluene (BHT), thiobarbituric acid, ergosterol, 1,3-dioleoyl-2-palmitoylglycerol, dioleoylglycerol, 2′, 7′-dichlorodihydrofluorescein diacetate (H₂DCFDA), and malondialdehyde (MDA) were purchased from Sigma-Aldrich (Poznan, Poland). Phospholipid standards were purchased from Avanti Polar Lipids (Alabaster, AL, USA). Sphingolipids standards were procured from Cayman Chemical (Ann Arbor, MI, USA). All other chemicals were acquired from Avantor Performance Materials (Gliwice, Poland). Stock solutions of 2,4-D were prepared at a concentration of 5 mg mL⁻¹ in ethanol.