Rapid quantification of conjugated and unconjugated bile acids and C27 precursors in dried blood spots and small volumes of serum[S]

Rapid quantification of conjugated and unconjugated bile acids and C27 precursors in dried blood spots and small volumes of serum[S]

The aim of the study was to develop a method for fast and reliable diagnosis of peroxisomal diseases and to facilitate differential diagnosis of cholestatic hepatopathy. For the quantification of bile acids and their conjugates as well as C27 precursors di- and trihydroxycholestanoic acid (DHCA, THC...

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Main Authors: N. Janzen, S. Sander, M. Terhardt, A.M. Das, J.O. Sass, R. Kraetzner, H. Rosewich, M. Peter, J. Sander
Format: Article
Language:English
Published: Elsevier 2010-06-01
Series:Journal of Lipid Research
Online Access:http://www.sciencedirect.com/science/article/pii/S002222752041034X
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spelling doaj-890d034771794a7a91ad815bff1e88332021-04-28T06:04:16ZengElsevierJournal of Lipid Research0022-22752010-06-0151615911598Rapid quantification of conjugated and unconjugated bile acids and C27 precursors in dried blood spots and small volumes of serum[S]N. Janzen0S. Sander1M. Terhardt2A.M. Das3J.O. Sass4R. Kraetzner5H. Rosewich6M. Peter7J. Sander8Department of Pediatric Kidney-, Liver-Metabolic Diseases, Children's Hospital Hannover Medical School, Hannover, Germany; Screening-Laboratory Hannover, Hannover, GermanyScreening-Laboratory Hannover, Hannover, GermanyScreening-Laboratory Hannover, Hannover, GermanyDepartment of Pediatric Kidney-, Liver-Metabolic Diseases, Children's Hospital Hannover Medical School, Hannover, GermanyLaboratory of Clinical Biochemistry and Metabolism, Centre for Pediatrics and Adolescent Medicine, University of Freiburg, GermanyDepartment of Pediatrics and Pediatric Neurology, Georg August University, Göttingen, GermanyDepartment of Pediatrics and Pediatric Neurology, Georg August University, Göttingen, GermanyScreening-Laboratory Hannover, Hannover, GermanyScreening-Laboratory Hannover, Hannover, GermanyThe aim of the study was to develop a method for fast and reliable diagnosis of peroxisomal diseases and to facilitate differential diagnosis of cholestatic hepatopathy. For the quantification of bile acids and their conjugates as well as C27 precursors di- and trihydroxycholestanoic acid (DHCA, THCA), in small pediatric blood samples we combined HPLC separation on a reverse-phase C18 column with ESI-MS/MS analysis in the negative ion mode. Analysis was done with good precision (CV 3,7%–11.1%) and sufficient sensitivity (LOQ: 11–91 nmol/L) without derivatization. Complete analysis of 17 free and conjugated bile acids from dried blood spots and 10 µL serum samples, respectively, was performed within 12 min. Measurement of conjugated primary bile acids plus DHCA and THCA as well as ursodeoxycholic acid was done in 4.5 min. In blood spots of healthy newborns, conjugated primary bile acids were found in the range of 0.01 to 2.01 µmol/L. Concentrations of C27 precursors were below the detection limit in normal controls. DHCA and THCA were specifically elevated in cases of peroxysomal defects and one Zellweger patient.http://www.sciencedirect.com/science/article/pii/S002222752041034X
collection DOAJ
language English
format Article
sources DOAJ
author N. Janzen
S. Sander
M. Terhardt
A.M. Das
J.O. Sass
R. Kraetzner
H. Rosewich
M. Peter
J. Sander
spellingShingle N. Janzen
S. Sander
M. Terhardt
A.M. Das
J.O. Sass
R. Kraetzner
H. Rosewich
M. Peter
J. Sander
Rapid quantification of conjugated and unconjugated bile acids and C27 precursors in dried blood spots and small volumes of serum[S]
Journal of Lipid Research
author_facet N. Janzen
S. Sander
M. Terhardt
A.M. Das
J.O. Sass
R. Kraetzner
H. Rosewich
M. Peter
J. Sander
author_sort N. Janzen
title Rapid quantification of conjugated and unconjugated bile acids and C27 precursors in dried blood spots and small volumes of serum[S]
title_short Rapid quantification of conjugated and unconjugated bile acids and C27 precursors in dried blood spots and small volumes of serum[S]
title_full Rapid quantification of conjugated and unconjugated bile acids and C27 precursors in dried blood spots and small volumes of serum[S]
title_fullStr Rapid quantification of conjugated and unconjugated bile acids and C27 precursors in dried blood spots and small volumes of serum[S]
title_full_unstemmed Rapid quantification of conjugated and unconjugated bile acids and C27 precursors in dried blood spots and small volumes of serum[S]
title_sort rapid quantification of conjugated and unconjugated bile acids and c27 precursors in dried blood spots and small volumes of serum[s]
publisher Elsevier
series Journal of Lipid Research
issn 0022-2275
publishDate 2010-06-01
description The aim of the study was to develop a method for fast and reliable diagnosis of peroxisomal diseases and to facilitate differential diagnosis of cholestatic hepatopathy. For the quantification of bile acids and their conjugates as well as C27 precursors di- and trihydroxycholestanoic acid (DHCA, THCA), in small pediatric blood samples we combined HPLC separation on a reverse-phase C18 column with ESI-MS/MS analysis in the negative ion mode. Analysis was done with good precision (CV 3,7%–11.1%) and sufficient sensitivity (LOQ: 11–91 nmol/L) without derivatization. Complete analysis of 17 free and conjugated bile acids from dried blood spots and 10 µL serum samples, respectively, was performed within 12 min. Measurement of conjugated primary bile acids plus DHCA and THCA as well as ursodeoxycholic acid was done in 4.5 min. In blood spots of healthy newborns, conjugated primary bile acids were found in the range of 0.01 to 2.01 µmol/L. Concentrations of C27 precursors were below the detection limit in normal controls. DHCA and THCA were specifically elevated in cases of peroxysomal defects and one Zellweger patient.
url http://www.sciencedirect.com/science/article/pii/S002222752041034X
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AT ssander rapidquantificationofconjugatedandunconjugatedbileacidsandc27precursorsindriedbloodspotsandsmallvolumesofserums
AT mterhardt rapidquantificationofconjugatedandunconjugatedbileacidsandc27precursorsindriedbloodspotsandsmallvolumesofserums
AT amdas rapidquantificationofconjugatedandunconjugatedbileacidsandc27precursorsindriedbloodspotsandsmallvolumesofserums
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