Enzymatic methods may underestimate the total serum bile acid concentration
Language English Country United States Media electronic-ecollection
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
32706797
PubMed Central
PMC7380613
DOI
10.1371/journal.pone.0236372
PII: PONE-D-20-09585
Knihovny.cz E-resources
- MeSH
- 3-alpha-Hydroxysteroid Dehydrogenase (B-Specific) chemistry MeSH
- Cholestasis metabolism MeSH
- Enzyme Assays methods MeSH
- Rats MeSH
- Humans MeSH
- Reproducibility of Results MeSH
- Bile Acids and Salts blood MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 3-alpha-Hydroxysteroid Dehydrogenase (B-Specific) MeSH
- Bile Acids and Salts MeSH
Enzymatic assays based on bacterial 3α-hydroxysteroid dehydrogenase are the method of choice for quantification of total bile acids (BAs) in serum. Although non-specific, it is generally considered precise and robust. The aim of this study was to investigate how changes in the BA spectrum might affect the reliability of the method. We measured standard solutions of twenty-three human and murine BAs using a commercial enzymatic assay and compared the measured vs. expected concentrations. Additionally, total BA concentrations in rat and human cholestatic samples with an abnormal BA spectrum were measured using an enzymatic assay, and a more specific LC-MS/MS method. We observed a great variability in the response of individual BAs in the enzymatic assay. Relative signal intensities ranged from 100% in glycocholic acid (reference) to only 20% in α-muricholic acid. The enzymatic assay markedly underestimated the BA concentrations in both human and rat cholestatic sera when compared to the LC-MS/MS assay. Our study indicated that the performance of an enzymatic assay largely depends on the BA spectrum, and the total concentration of BAs can be markedly underestimated. Samples with an atypical BA spectrum (viz. in rodents) should preferably be measured by other methods.
4th Department of Internal Medicine 1st Faculty of Medicine Charles University Prague Czech Republic
See more in PubMed
European Association for the Study of the Liver. EASL Clinical Practice Guidelines: Management of cholestatic liver diseases. Journal of Hepatology. 2009;51(2):237–67. 10.1016/j.jhep.2009.04.009 PubMed DOI
Chiang JYL. Bile acid metabolism and signaling. Compr Physiol. 2013;3(3):1191–212. 10.1002/cphy.c120023 . PubMed DOI PMC
Chiang JYL, Ferrell JM. Bile Acids as Metabolic Regulators and Nutrient Sensors. Annual Review of Nutrition. 2019;39(1):175–200. 10.1146/annurev-nutr-082018-124344 . PubMed DOI PMC
Iwata T, Yamasaki K. Enzymatic Determination and Thin-layer Chromatography of Bile Acids in Blood. The Journal of Biochemistry. 1964;56(5):424–31. PubMed
Mashige F, Tanaka N, Maki A, Kamei S, Yamanaka M. Direct spectrophotometry of total bile acids in serum. Clinical Chemistry. 1981;27(8):1352 PubMed
Muchova L, Vanova K, Suk J, Micuda S, Dolezelova E, Fuksa L, et al. Protective effect of heme oxygenase induction in ethinylestradiol-induced cholestasis. Journal of cellular and molecular medicine. 2015;19(5):924–33. Epub 2015/02/16. 10.1111/jcmm.12401 . PubMed DOI PMC
Prasnicka A, Cermanova J, Hroch M, Dolezelova E, Rozkydalova L, Smutny T, et al. Iron depletion induces hepatic secretion of biliary lipids and glutathione in rats. Biochimica et Biophysica Acta (BBA)—Molecular and Cell Biology of Lipids. 2017;1862(12):1469–80. 10.1016/j.bbalip.2017.09.003. PubMed DOI
Engert R, Turner MD. Problems in the measurement of bile acids with 3α-hydroxysteroid dehydrogenase. Analytical Biochemistry. 1973;51(2):399–407. 10.1016/0003-2697(73)90493-4 PubMed DOI
Danese E, Salvagno GL, Negrini D, Brocco G, Montagnana M, Lippi G. Analytical evaluation of three enzymatic assays for measuring total bile acids in plasma using a fully-automated clinical chemistry platform. PloS one. 2017;12(6):e0179200–e. 10.1371/journal.pone.0179200 . PubMed DOI PMC
Trottier J, Bialek A, Caron P, Straka RJ, Milkiewicz P, Barbier O. Profiling Circulating and Urinary Bile Acids in Patients with Biliary Obstruction before and after Biliary Stenting. Plos One. 2011;6(7). PubMed PMC
Ye L, Liu SY, Wang M, Shao Y, Ding M. High-performance liquid chromatography-tandem mass spectrometry for the analysis of bile acid profiles in serum of women with intrahepatic cholestasis of pregnancy. Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences. 2007;860(1):10–7. PubMed
Galloway D, Mezoff E, Zhang W, Byrd M, Cole C, Aban I, et al. Serum Unconjugated Bile Acids and Small Bowel Bacterial Overgrowth in Pediatric Intestinal Failure: A Pilot Study. Journal of Parenteral and Enteral Nutrition. 2019;43(2):263–70. 10.1002/jpen.1316 PubMed DOI PMC
