Hyflo super cel

Potassium persulfate (K₂S₂O₈), AAPH (2,2′-azobis-2-methyl-propanimidamide), fluorescein sodium salt, Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), ABTS (2,2′-azinobis (3-ethylbenzothiazoline 6-sulfonic acid), diatomaceous earth (Hyflo^®Super Cel^®), and astaxanthin standard were bought from Sigma-Aldrich (Steinheim, Baden-Württemberg, Germany). Dimethyl sulfoxide (DMSO) and absolute ethanol were provided by VWR International Eurolab S.L. (Barcelona, Spain).

Total fat content was quantified by Soxhlet extraction with petroleum ether following the procedure described by the ISO 12966-2:2017: preparation of methyl esters of fatty acids, animal and vegetable fats, and oils [22 ]. Gas chromatography of fatty acid methyl esters was employed for the analysis of the individual profile. The fat content and fatty acids methyl ester profile are in g·kg⁻¹ of dry matter.
Petroleum ether 40–65 °C (Sigma-Aldrich, Steinheim, Germany), filter medium Hyflo Super Cel (diatomaceous earth) (Sigma-Aldrich, Steinheim, Germany), methanol for chromatography (Sigma-Aldrich, Steinheim, Germany), KOH (Centralchem, Bratislava, Slovakia), HCl 33–36% (Centralchem, Bratislava, Slovakia), NaHSO₄ H₂O (Centralchem, Bratislava, Slovakia), Na₂SO₄ (Centralchem, Bratislava, Slovakia), anhydrous N-hexane for chromatography (Sigma-Aldrich, Steinheim Germany), 37 component standard FAME mix 10 mg.mL⁻¹ in methylene chloride containing C4-C24 FAME (2–4% relative concentration), manufacturer Supelco, catalog number 47885-U chromatography (Sigma-Aldrich, Steinheim, Germany).
1. Tecator Soxtec System HT 1043 Extraction unit (Gemini, Apeldoom, the Netherlands) apparatus at a flow rate of 10 cycles/h or 10 mL·min⁻¹;
2. Vacuum oven, (Fisher Scientific, Waltham, MA, USA) temperature adjustable (100 ± 3 °C).

Hyflo Super Cel (Diatomaceous earth), 3-aminopropyl(diethoxy)methylsilane (APDMS, 97%), dimethyl suberimidate dihydrochloride (DMS, 98%), lysozyme solution (50 mg/mL in distilled water), sodium hydroxide solution (50% in H₂O), N-Acetyl-L-cysteine (NALC, 99%), sodium citrate, and Triton X-100 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Tris-HCI (pH 8.0), distilled water (DNase/RNase-Free), and EDTA (pH 8.0) were purchased from Invitrogen (Waltham, MA, USA). Proteinase K solution (>600 mAU/mL) was purchased from Qiagen (Hilden, Germany). Absolute ethanol was purchased from Merck (Whitehouse Station, NJ, USA). Phosphate-buffered saline (PBS; 10×, pH 7.4) was purchased from Gibco (Grand Island, NY, USA).