F002 1 1

Contents of moisture, crude protein, IMF, ash, and inosinic acid in LT were measured by freeze-dryer (ALPHA 2–4 LSC, Christ Martin GmbH), automatic nitrogen analyzer (8400, FOSS, Denmark), fat analyzer (2055 SOXTEC, FOSS, Denmark), box resistance furnace (SX2-4-10N, Yiheng, Shanghai, China), and high performance liquid chromatography (HPLC, LC-20AD, Shimazu, Japan), respectively, as described previously (16 (link)). Briefly, the contents of moisture, crude protein, IMF, and ash were analyzed by the freeze-drying method, Kjeldahl method, Soxhlet extraction, and burning method, respectively.
The sample preparation procedure comprised homogenization of LT (0.1 g) and 0.9% saline (0.9 mL), and 1,500 r/min for 2 min, followed by centrifugation at 3,500 × g for 15 min at 4°C to collect the supernatant. Then, contents of cholesterol, triglyceride, and malondialdehyde (MDA) and activities of total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) were determined according to the instructions (F001-1-1, F002-1-1, A003-2-2, A015-1-2, A005-1-2, A001-1-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China). The plate was read by a multi-functional enzyme labeling instrument (Spectra Max M5, Molecular Devices, USA) at 532 nm (MDA), 520 nm (T-AOC), 412 nm (GSH-Px), and 450 nm (SOD).

Blood samples were collected and left for 2–3 h at room temperature. The serum was centrifuged at 3000 rpm for 15 min, divided, and stored at −80 °C. TC, TG, LDL-C, and HDL-C levels in serum were determined using commercially available assay kits (cat: F002-1-1, F001-1-1, A113-2-1, and A112-2-1, Nanjing Jiancheng Bioengineering Institute, China).

TC, TG, FFA, LDL-C, and HDL-C levels were measured by corresponding kits according to the manufacturer’s instructions (F001-1-1, F002-1-1, A042-2-1, A112-1-1, and A113-1-1; Jiancheng Bioengineering Institute, Nanjing, China).