Tba test

Dried feces (10 mg) were dissolved in 0.2 mL 90% ethanol and incubated at 65 °C for 1 h. The samples were then dissolved in 0.5 mL 90% ethanol. The total levels of bile acids in the plasma and feces were measured with the TBA-Test (Wako). Each liver tissue sample (0.1 g) was treated with Folch solution and homogenized. Total levels of cholesterol were measured by the T-cho E-Test (Wako). Extracted lipids were quantitatively analyzed. As a recovery standard, norcholic acid (Steraloids, Inc., Newport, RI; 1 mg/mL stock solution in 50% ethanol) was added to the ethanol. Filtrated extracts were centrifuged at 12,000 × g for 5 min and diluted 10 times in water. The diluted extracts were then analyzed using high-pressure liquid chromatography-tandem mass spectrometry (50 mm × 1 mm C8 column, buffered mobile phase) [32 (link)].

The blood triglyceride, cholesterol, glucose, non-esterified fatty acid (NEFA) and bile acid concentrations were measured using commercial kits (triglyceride E-test, T-cholesterol E-test, glucose CII-test, NEFA-C test and TBA-test; Wako Pure Chemical Industries, Osaka, Japan). The insulin and corticosterone levels were determined using ELISA kits (rat insulin ELISA kit, Morinaga Institute of Biological Science, Yokohama, Japan; corticosterone ELISA kit, Enzo life sciences, NY).
About 2.5 g of liver was homogenized and lipids extracted, following the method described by Folch et al. [16 (link)]. Total lipids in the liver were determined gravimetrically. The concentrations of hepatic triglycerides, cholesterol, and phospholipid in the extracts were measured using commercial kits (phospholipids C-test: Wako Pure Chemical Industries, Osaka, Japan).

We used commercial kits to measure plasma concentrations of glucose (Glucose CII-test, Wako, Osaka, Japan), TGs (TG E-test, Wako), non-esterified free fatty acids (NEFA C-test, Wako, Osaka, Japan), total bile acids (TBA-test, Wako, Osaka, Japan), total cholesterol (cholesterol E-test, Wako, Osaka, Japan), insulin (Insulin ELISA; Shibayagi; Gunma, Japan), and corticosterone (YK240; Yanaihara Institute, Inc.; Shizuoka, Japan).