Bile Acids Quantification by Liquid Chromatography-Tandem Mass Spectrometry: Method Validation, Reference Range, and Interference Study
| Autor: | Davide Ambrogi, Davide Negrini, Patricia M.-J. Lievens, Simone Donzelli, Giuseppe Lippi, Mairi Pucci, Elisa Danese, Simone De Nitto, Gian Luca Salvagno |
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| Rok vydání: | 2020 |
| Předmět: |
Analyte
LC–MS/MS bile acids interference Bilirubin Clinical Biochemistry interference Reference range Article 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cholestasis LC–MS/MS In vivo Liquid chromatography–mass spectrometry medicine 030304 developmental biology bile acids lcsh:R5-920 0303 health sciences Chromatography Chemistry Jaundice medicine.disease 030220 oncology & carcinogenesis Hemoglobin medicine.symptom lcsh:Medicine (General) |
| Zdroj: | Diagnostics Diagnostics, Vol 10, Iss 462, p 462 (2020) Volume 10 Issue 7 |
| ISSN: | 2075-4418 |
| Popis: | Bile acids (BA) play a pivotal role in cholesterol metabolism. Their blood concentration has also been proposed as new prognostic and diagnostic indicator of hepatobiliary, intestinal, and cardiovascular disease. Liquid chromatography tandem mass spectrometry (LC&ndash MS/MS) currently represents the gold standard for analysis of BA profile in biological samples. We report here development and validation of a LC&ndash MS/MS technique for simultaneously quantifying 15 BA species in serum samples. We also established a reference range for adult healthy subjects (n = 130) and performed a preliminary evaluation of in vitro and in vivo interference. The method displayed good linearity, with high regression coefficients (> 0.99) over a range of 5 ng/mL (lower limit of quantification, LLOQ) and 5000 ng/mL for all analytes tested. The accuracies were between 85&ndash 115%. Both intra- and inter-assay imprecision was < 10%. The recoveries ranged between 92&ndash 110%. Each of the tested BA species (assessed on three concentrations) were stable for 15 days at room temperature, 4 ° C, and &minus 20 ° C. The in vitro study did not reveal any interference from triglycerides, bilirubin, or cell-free hemoglobin. The in vivo interference study showed that pools obtained from hyper-cholesterolemic patients and hyper-bilirubinemic patients due to post-hepatic jaundice for benign cholestasis, cholangiocarcinoma and pancreatic head tumors had clearly distinct patterns of BA concentrations compared with a pool obtained from samples of healthy subjects. In conclusion, this study proposes a new suitable candidate method for identification and quantitation of BA in biological samples and provides new insight into a number of variables that should be taken into account when investigating pathophysiological changes of BA in human diseases. |
| Databáze: | OpenAIRE |
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