Triglycerides lq gpo pod

Feces were dried at 45 °C for 24 h prior to triglyceride extraction. Dried feces (200 mg) were mixed with 2 mL of 0.9% NaCl and with 2 mL of 2:1 (v/v) chloroform:methanol. Next, samples were centrifuged at 6000× g for 5 min at 25 °C, and 1 mL of the lower phase was collected in a new tube. Then, samples were vacuum dried at 35 °C and resuspended in 200 μL of ethanol. Finally, following the manufacturer’s instructions, triglycerides were determined using a commercial enzymatic-colorimetric kit (Triglycerides-LQ GPO-POD, Spinreact). Results were expressed as mg of triglycerides/g of feces [11 (link)].
Frozen livers (200 mg) were digested with 350 μL of 30% potassium hydroxide in ethanol (2:1, v/v) at 55 °C overnight. Then, 650 μL water:ethanol (1:1, v/v) was added, and samples were centrifuged at 6000× g for 5 min. Supernatants (200 μL) were recovered and mixed with 215 μL of 1 M magnesium chloride. Samples were incubated in ice for 10 min and centrifuged at 6000× g. Finally, following the manufacturer’s instructions, triglycerides were determined using a commercial enzymatic-colorimetric kit (Triglycerides-LQ GPO-POD, Spinreact). Results were expressed as mg of triglycerides/g of the liver [12 (link)].

Serum glucose (Glucose-LQ GOD-POD, Spinreact, Girone, Spain), triglycerides (Triglycerides-LQ GPO-POD, Spinreact), aspartate transaminase (GOT-AST-LQ, Spinreact), and alanine transaminase (GPT-ALT-LQ, Spinreact) were determined using commercial enzymatic-colorimetric kits following the manufacturer’s instructions. Following the manufacturers’ instructions, serum insulin was determined using an ELISA kit (EZRMI-13K, EMD Millipore Co., Burlington, MA, USA). Insulin resistance- and beta-Homeostatic model assessment (HOMA-IR and HOMA-beta), quantitative insulin sensitivity check index (QUICKI), fasting glucose-to-insulin ratio (FGIR), and fasting triglycerides and glucose (TyG) indexes were calculated with the following equations: $$\mathrm{HOMA}-\mathrm{IR}=\frac{\mathrm{glucose} \left(\raisebox{1ex}{$\mathrm{mg}$}\!\left/ \!\raisebox{-1ex}{$\mathrm{dL}$}\right.\right)\times \mathrm{insulin}\left(\raisebox{1ex}{$\mathsf{\mu }\mathrm{M}$}\!\left/ \!\raisebox{-1ex}{$\mathrm{mL}$}\right.\right)}{2430}$$
$$\mathrm{HOMA}-\mathrm{Beta}=\frac{\mathrm{insulin} \left(\raisebox{1ex}{$\mathrm{ng}$}\!\left/ \!\raisebox{-1ex}{$\mathrm{mL}$}\right.\right)\times 360}{\mathrm{glucose} \left(\raisebox{1ex}{$\mathrm{mg}$}\!\left/ \!\raisebox{-1ex}{$\mathrm{dL}$}\right.\right)-63}$$
$$\mathrm{QUICKI}=\frac{1}{\log \left[\mathrm{insulin}\left(\raisebox{1ex}{$\mathsf{\mu }\mathrm{M}$}\!\left/ \!\raisebox{-1ex}{$\mathrm{mL}$}\right.\right)\right]+\log \left[\mathrm{glucose}\left(\raisebox{1ex}{$\mathrm{mg}$}\!\left/ \!\raisebox{-1ex}{$\mathrm{dL}$}\right.\right)\right]}$$
$$\mathrm{FGIR}=\frac{\mathrm{glucose} \left(\raisebox{1ex}{$\mathrm{mg}$}\!\left/ \!\raisebox{-1ex}{$\mathrm{dL}$}\right.\right)}{\mathrm{insulin} \left(\raisebox{1ex}{$\mathsf{\mu }\mathrm{M}$}\!\left/ \!\raisebox{-1ex}{$\mathrm{mL}$}\right.\right)}$$
$$\mathrm{TyG}=\frac{\ln \left[\mathrm{glucose}\left(\raisebox{1ex}{$\mathrm{mg}$}\!\left/ \!\raisebox{-1ex}{$\mathrm{dL}$}\right.\right)\ast \mathrm{triglycerides}\left(\raisebox{1ex}{$\mathrm{mg}$}\!\left/ \!\raisebox{-1ex}{$\mathrm{dL}$}\right.\right)\right]}{2}$$

Total plasma triglycerides and cholesterol concentrations were analyzed with a microtiter assay using Triglycerides-LQ GPO-POD and Cholesterol-LQ CHOD-POD (Spinreact) commercial kits. For plasma lipoprotein profiles, 400 µL of pooled plasmas from each group were analysed using fast phase liquid chromatography (FPLC) method. Separation was performed by gel filtration using a Superose 6 h 10/30 column (Pharmacia) as previously described^{59 (link)}.