Plasma alanine aminotransferase (ALT), alkaline phosphatase, and total bilirubin were quantified using DRI-CHEM NX500 (Fuji Film RI Pharma, Tokyo, Japan). BA concentrations and profiles in plasma, liver, and gallbladder bile were evaluated with a Total Bile Acid-Test Wako Kit (FUJIFILM Wako Pure Chemical) and an liquid chromatography with tandem mass spectrometry system.19 (link) To analyze the liver, 100 μg liver sections were homogenized in 400 μL 100% ethanol by Tissue Lyser II (QIAGEN, Hilden, Germany) and centrifuged at 1700 × g for 5 min at 4°C. The supernatants were collected, concentrated with centrifugal thickener for 2 h, resuspended in 100 μL 80% ethanol, and subjected to BA analysis. The hydrophobicity index of BAs was calculated as previously.20 (link)
Total bile acid test wako kit
Manufactured by Fujifilm
Sourced in Germany
The Total Bile Acid-Test Wako Kit is a laboratory equipment product designed for the quantitative determination of total bile acids in biological samples. It provides a reliable and accurate method for measuring the concentration of bile acids in various matrices such as serum, plasma, or other relevant samples.
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2 protocols using total bile acid test wako kit
1
Bile Acid Profiling in Liver Disease
2
Biochemical Analysis of Sera and Liver Lipids
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The biochemical parameters of sera were analyzed using a commercial service (Japan Medical Laboratory, Osaka, Japan). The total lipid of liver samples were extracted using the chloroform/methanol/water method (16 (link)). Each extracted total lipid fraction was dissolved in 10 volumes of ethanol, and cholesterol and TAG contents were measured using commercial kits (Cholesterol E-test and Triglyceride E-test, Wako Pure Chemical Industries, Ltd., Osaka, Japan). Phospholipid contents in extracted liver lipids were estimated by phosphorus analyses (22 (link)).
Fatty acids content in feces samples was analyzed in accordance with a previous study (23 (link)). The neutral sterol (sum of coprostanol and cholesterol) content in feces samples was analyzed by GC (GC-2014, Shimadzu Corporation) as described above. The acidic sterol content in feces was measured with a commercial kit (Total Bile Acid Test Wako kit, Wako Pure Chemical Industries, Ltd.) according to the manufacturer’s protocol. Nitrogen contents in feces were determined by the Micro-Kjeldahl method as described above.
Fatty acids content in feces samples was analyzed in accordance with a previous study (23 (link)). The neutral sterol (sum of coprostanol and cholesterol) content in feces samples was analyzed by GC (GC-2014, Shimadzu Corporation) as described above. The acidic sterol content in feces was measured with a commercial kit (Total Bile Acid Test Wako kit, Wako Pure Chemical Industries, Ltd.) according to the manufacturer’s protocol. Nitrogen contents in feces were determined by the Micro-Kjeldahl method as described above.
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