Serum bile acids profiling by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and its application on pediatric liver and intestinal diseases

Author:

Fu Xiaowei1,Xiao Yi2,Golden Jamie3,Niu Sizhe4,Gayer Christopher P.3

Affiliation:

1. Department of Pathology and Laboratory Medicine, LeBonheur Children’s Hospital, University of Tennessee Health Science Center, 50 N Dunlap St, Memphis, TN 38103, USA, E-mail: xiaoweifu@hotmail.com

2. Department of Pathology and Laboratory Medicine, Children’s Hospital of Los Angeles, Los Angeles, CA, USA

3. Division of Pediatric Surgery, Children’s Hospital Los Angeles, Los Angeles, CA, USA

4. Department of Pediatrics, Jan and Dan Duncan Neurological Research Institute, Texas Children’s Hospital, Baylor College of Medicine, Houston, TX, USA

Abstract

AbstractBackgroundA method for bile acid profiling measuring 21 primary and secondary bile acids in serum samples was developed and validated with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Sample preparation included spiking with internal standards followed by protein precipitation, centrifugation, drying under nitrogen gas and reconstitution. Extracted samples were injected onto a Phenomenex Kinetex C18 column (150 × 4.60 mm, 2.6 μm).MethodsData was collected with LC-MS/MS operated in negative ion mode with multiple reaction monitoring (MRM) and single reaction monitoring (SRM). The analytical run time was 12 min.ResultsThe method showed excellent linearity with high regression coefficients (>0.99) over a range of 0.05 and 25 μM for all analytes tested. The method also showed acceptable intra-day and inter-day accuracy and precision. As a proof of concept, the analytical method was applied to patients with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), biliary atresia (BA), and necrotizing enterocolitis (NEC), and distinct bile acids profiles were demonstrated.ConclusionsThe method could be poised to identify possible biomarkers for non-invasive early diagnosis of these disorders.

Publisher

Walter de Gruyter GmbH

Subject

Biochemistry, medical,Clinical Biochemistry,General Medicine

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