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Simple enzymatic assay for determining cholesterol concentrations in bile.
C M Luhman1, S T Galloway, D C Beitz
1Department of Animal Science, Iowa State University, Ames 50011.
Clinical Chemistry
|February 1, 1990
Summary
Bilirubin oxidase enzyme removes bilirubin interference in cholesterol assays for bile. This method accurately measures cholesterol and avoids issues with other enzymatic assays, improving accuracy for biliary constituents.
Area of Science:
- Biochemistry
- Analytical Chemistry
Background:
- Bilirubin interference complicates enzymatic assays for cholesterol in bile.
- Existing methods are often affected by chromogen oxidase activity.
Purpose of the Study:
- To develop a method for accurate cholesterol measurement in bile by eliminating bilirubin interference.
- To utilize bilirubin oxidase for improved enzymatic assays.
Main Methods:
- Enzymatic assay using bilirubin oxidase (EC 1.3.3.5) to convert bilirubin to biliverdin.
- Samples treated with bilirubin oxidase for 10 minutes prior to cholesterol reagent addition.
- Use of sample blanks to correct for residual biliverdin interference.
Main Results:
- Accurate and precise determination of cholesterol concentration in bile was achieved.
- The method is simple, convenient, and avoids "chromogen oxidase" activity issues.
- Residual biliverdin interference was effectively managed using sample blanks.
Conclusions:
- Bilirubin oxidase is effective in removing bilirubin interference for cholesterol assays in bile.
- This approach offers a reliable and improved method for analyzing biliary constituents.
- The technique is potentially applicable to other biological samples like urine and plasma.