Automatic biochemical analyzer

The in vitro ability of polysaccharides to bind bile acids in a simulated intestinal environment was determined according to the previously reported method with some modifications [25 (link)]. Briefly, 1 mL of the polysaccharide solution (0.2, 0.5 and 1.0 mg/mL) and 1.25 mL of 0.01 mol/L HCl were mixed and incubated at 37 °C for 1 h. After this acidic incubation, which simulated gastric digestion, the pH of sample was adjusted to 6.3. During the next step, 2 mL of the bile acids solution (0.72 μmol/mL) was added and mixed, followed by 2.5 mL of porcine pancreatin (10 mg/mL) that provided amylase, protease and lipase for digestion. This was subsequently incubated for 1 h at 37 °C in shaker bath. The mixtures were transferred to 10-mL centrifuge tubes and centrifuged at 4800× g for 10 min at 25 °C. To determine the total amount of bile acids bound by the polysaccharides, the final bile acids content in the supernatant was measured. The bile acids were measured by an Automatic Biochemical Analyzer (AU2700, Olympus, Hamburg, Germany). Values were determined from a standard curve obtained using the tested bile acids solution. Cellulose, a non bile acid binding fiber, was the negative control and cholestyramine, a bile acid binding anionic resin, was the positive control.

Aspartate transaminase (AST), lactate dehydrogenase (LDH), creatine kinase (CK), and creatine kinase-MB (CK-MB) and the levels of total cholesterol (TC) and triglyceride (TG) in the serum were measured using an automatic biochemical analyzer (Olympus, Tokyo, Japan).

Quantitative determination of BUN and Cr of the rats was done on Automatic Biochemical Analyzer (Olympus AU5400, Tokyo, Japan).