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Preanalytical Biases in the Measurement of Human Blood Sphingolipids

Dysregulation of blood sphingolipids is an emerging topic in clinical science. The objective of this study was to determine preanalytical biases that typically occur in clinical and translational studies and that influence measured blood sphingolipid levels. Therefore, we collected blood samples fro...

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Autores principales: Brunkhorst, Robert, Pfeilschifter, Waltraud, Patyna, Sammy, Büttner, Stefan, Eckes, Timon, Trautmann, Sandra, Thomas, Dominique, Pfeilschifter, Josef, Koch, Alexander
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983773/
https://www.ncbi.nlm.nih.gov/pubmed/29735920
http://dx.doi.org/10.3390/ijms19051390
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author Brunkhorst, Robert
Pfeilschifter, Waltraud
Patyna, Sammy
Büttner, Stefan
Eckes, Timon
Trautmann, Sandra
Thomas, Dominique
Pfeilschifter, Josef
Koch, Alexander
author_facet Brunkhorst, Robert
Pfeilschifter, Waltraud
Patyna, Sammy
Büttner, Stefan
Eckes, Timon
Trautmann, Sandra
Thomas, Dominique
Pfeilschifter, Josef
Koch, Alexander
author_sort Brunkhorst, Robert
collection PubMed
description Dysregulation of blood sphingolipids is an emerging topic in clinical science. The objective of this study was to determine preanalytical biases that typically occur in clinical and translational studies and that influence measured blood sphingolipid levels. Therefore, we collected blood samples from four healthy male volunteers to investigate the effect of storage conditions (time, temperature, long-term storage, freeze–thaw cycles), blood drawing (venous or arterial sampling, prolonged venous compression), and sample preparation (centrifugation, freezing) on sphingolipid levels measured by LC-MS/MS. Our data show that sphingosine 1-phosphate (S1P) and sphinganine 1-phosphate (SA1P) were upregulated in whole blood samples in a time- and temperature-dependent manner. Increased centrifugation at higher speeds led to lower amounts of S1P and SA1P. All other preanalytical biases did not significantly alter the amounts of S1P and SA1P. Further, in almost all settings, we did not detect differences in (dihydro)ceramide levels. In summary, besides time-, temperature-, and centrifugation-dependent changes in S1P and SA1P levels, sphingolipids in blood remained stable under practically relevant preanalytical conditions.
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spelling pubmed-59837732018-06-05 Preanalytical Biases in the Measurement of Human Blood Sphingolipids Brunkhorst, Robert Pfeilschifter, Waltraud Patyna, Sammy Büttner, Stefan Eckes, Timon Trautmann, Sandra Thomas, Dominique Pfeilschifter, Josef Koch, Alexander Int J Mol Sci Article Dysregulation of blood sphingolipids is an emerging topic in clinical science. The objective of this study was to determine preanalytical biases that typically occur in clinical and translational studies and that influence measured blood sphingolipid levels. Therefore, we collected blood samples from four healthy male volunteers to investigate the effect of storage conditions (time, temperature, long-term storage, freeze–thaw cycles), blood drawing (venous or arterial sampling, prolonged venous compression), and sample preparation (centrifugation, freezing) on sphingolipid levels measured by LC-MS/MS. Our data show that sphingosine 1-phosphate (S1P) and sphinganine 1-phosphate (SA1P) were upregulated in whole blood samples in a time- and temperature-dependent manner. Increased centrifugation at higher speeds led to lower amounts of S1P and SA1P. All other preanalytical biases did not significantly alter the amounts of S1P and SA1P. Further, in almost all settings, we did not detect differences in (dihydro)ceramide levels. In summary, besides time-, temperature-, and centrifugation-dependent changes in S1P and SA1P levels, sphingolipids in blood remained stable under practically relevant preanalytical conditions. MDPI 2018-05-07 /pmc/articles/PMC5983773/ /pubmed/29735920 http://dx.doi.org/10.3390/ijms19051390 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Brunkhorst, Robert
Pfeilschifter, Waltraud
Patyna, Sammy
Büttner, Stefan
Eckes, Timon
Trautmann, Sandra
Thomas, Dominique
Pfeilschifter, Josef
Koch, Alexander
Preanalytical Biases in the Measurement of Human Blood Sphingolipids
title Preanalytical Biases in the Measurement of Human Blood Sphingolipids
title_full Preanalytical Biases in the Measurement of Human Blood Sphingolipids
title_fullStr Preanalytical Biases in the Measurement of Human Blood Sphingolipids
title_full_unstemmed Preanalytical Biases in the Measurement of Human Blood Sphingolipids
title_short Preanalytical Biases in the Measurement of Human Blood Sphingolipids
title_sort preanalytical biases in the measurement of human blood sphingolipids
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983773/
https://www.ncbi.nlm.nih.gov/pubmed/29735920
http://dx.doi.org/10.3390/ijms19051390
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