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Updated: Jul 15, 2026
Dried Blood Spot Collection of Health Biomarkers to Maximize Participation in Population Studies
Published on: January 28, 2014
Differences in bile acid profiles between cholestatic diseases - Development of a high throughput assay for dried
Anders Ziegler1, Ingjerd Sæves2, Runar Almaas3
1Department of Pediatric Research, Oslo University Hospital, Sognsvannsveien 20, 0372 Oslo, Norway; Department of Newborn Screening, Oslo University Hospital, Forskningsveien 2B, 0373 Oslo, Norway; Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway.
Background:
Cholestasis causes accumulation of bile acids (BAs) and changes the circulating bile acid profile. Quantification of circulating BAs in dried bloodspots (DBS) may demonstrate obstruction of bile flow and altered bile acid metabolism in the liver. High sample throughput enables rapid screening of cholestatic diseases.
Materials And Methods:
Ultra high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used for optimizing separation and detection of the primary unconjugated BAs cholic acid (CA) and chenodeoxycholic acid (CDCA); the secondary unconjugated BAs ursodeoxycholic acid (UDCA), hyodeoxycholic acid (HDCA) and deoxycholic acid (DCA), as well as the glycine- and taurine-conjugated variants of CA, CDCA, DCA and UDCA. Donor blood was obtained to prepare DBS calibrators and quality controls for method development and validation.
Results:
We developed a quantitative bile acid assay with a run-time of two minutes, and one-step sample preparation of 3.2 mm DBS discs. Validation results demonstrated overall good performance and was considered fit for purpose. Children with Alagille syndrome, Aagenaes syndrome and alpha-1 antitrypsin deficiency had increased BAs in DBS from newborn screening samples compared with age matched controls, and had different bile acids profiles.
Conclusion:
We propose that our high throughput assay allows bile acid profiling in DBS that can be a valuable assessment tool for early screening of cholestasis in children. Assaying BAs in dried bloodspots is key for early detection of cholestasis, and provides transferability to a newborn screening setting.
