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Quantification of sphingosine 1-phosphate by validated LC-MS/MS method revealing strong correlation with apolipoprotein M in plasma but not in serum due to platelet activation during blood coagulation.

Frej, Cecilia LU ; Andersson, Anders S LU ; Larsson, Benny; Guo, Li Jun LU ; Norström, Eva LU ; Happonen, Kaisa LU and Dahlbäck, Björn LU (2015) In Analytical and Bioanalytical Chemistry 407(28). p.8533-8542
Abstract
Sphingosine 1-phosphate (S1P) is a signalling sphingolipid affecting multiple cellular functions of vascular and immune systems. It circulates at submicromolar levels bound to HDL-associated apolipoprotein M (apoM) or to albumin. S1P in blood is mainly produced by platelets and erythrocytes, making blood sampling for S1P quantification delicate. Standardisation of sampling is thereby of great importance to obtain robust data. By optimising and characterising the extraction procedure and the LC-MS/MS analysis, we have developed and validated a highly specific and sensitive method for S1P quantification. Blood was collected from healthy individuals (n = 15) to evaluate the effects of differential blood sampling on S1P levels. To evaluate... (More)
Sphingosine 1-phosphate (S1P) is a signalling sphingolipid affecting multiple cellular functions of vascular and immune systems. It circulates at submicromolar levels bound to HDL-associated apolipoprotein M (apoM) or to albumin. S1P in blood is mainly produced by platelets and erythrocytes, making blood sampling for S1P quantification delicate. Standardisation of sampling is thereby of great importance to obtain robust data. By optimising and characterising the extraction procedure and the LC-MS/MS analysis, we have developed and validated a highly specific and sensitive method for S1P quantification. Blood was collected from healthy individuals (n = 15) to evaluate the effects of differential blood sampling on S1P levels. To evaluate correlation between S1P and apoM in different types of plasma and serum, apoM was measured by ELISA. The method showed good accuracy and precision in the range of 0.011 to 0.9 μM with less than 0.07 % carryover. We found that the methanol precipitation used to extract S1P co-extracted apoM and several other HDL-proteins from plasma. The platelet-associated S1P was released during coagulation, thus increasing the S1P concentration to double in serum as compared to that in plasma. Gel filtration chromatography revealed that the platelet-released S1P was mainly bound to albumin. This explains why the strong correlation between S1P and apoM levels in plasma is lost upon the clotting process and hence not observed in serum. We have developed, characterised and validated an efficient, highly sensitive and specific method for the quantification of S1P in biological material. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical and Bioanalytical Chemistry
volume
407
issue
28
pages
8533 - 8542
publisher
Springer
external identifiers
  • pmid:26377937
  • wos:000365168900015
  • scopus:84941695034
ISSN
1618-2642
DOI
10.1007/s00216-015-9008-4
language
English
LU publication?
yes
id
5810b51a-c1e5-471b-964f-c12ed710054d (old id 8038954)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/26377937?dopt=Abstract
date added to LUP
2015-10-03 19:57:36
date last changed
2017-09-17 03:44:25
@article{5810b51a-c1e5-471b-964f-c12ed710054d,
  abstract     = {Sphingosine 1-phosphate (S1P) is a signalling sphingolipid affecting multiple cellular functions of vascular and immune systems. It circulates at submicromolar levels bound to HDL-associated apolipoprotein M (apoM) or to albumin. S1P in blood is mainly produced by platelets and erythrocytes, making blood sampling for S1P quantification delicate. Standardisation of sampling is thereby of great importance to obtain robust data. By optimising and characterising the extraction procedure and the LC-MS/MS analysis, we have developed and validated a highly specific and sensitive method for S1P quantification. Blood was collected from healthy individuals (n = 15) to evaluate the effects of differential blood sampling on S1P levels. To evaluate correlation between S1P and apoM in different types of plasma and serum, apoM was measured by ELISA. The method showed good accuracy and precision in the range of 0.011 to 0.9 μM with less than 0.07 % carryover. We found that the methanol precipitation used to extract S1P co-extracted apoM and several other HDL-proteins from plasma. The platelet-associated S1P was released during coagulation, thus increasing the S1P concentration to double in serum as compared to that in plasma. Gel filtration chromatography revealed that the platelet-released S1P was mainly bound to albumin. This explains why the strong correlation between S1P and apoM levels in plasma is lost upon the clotting process and hence not observed in serum. We have developed, characterised and validated an efficient, highly sensitive and specific method for the quantification of S1P in biological material.},
  author       = {Frej, Cecilia and Andersson, Anders S and Larsson, Benny and Guo, Li Jun and Norström, Eva and Happonen, Kaisa and Dahlbäck, Björn},
  issn         = {1618-2642},
  language     = {eng},
  number       = {28},
  pages        = {8533--8542},
  publisher    = {Springer},
  series       = {Analytical and Bioanalytical Chemistry},
  title        = {Quantification of sphingosine 1-phosphate by validated LC-MS/MS method revealing strong correlation with apolipoprotein M in plasma but not in serum due to platelet activation during blood coagulation.},
  url          = {http://dx.doi.org/10.1007/s00216-015-9008-4},
  volume       = {407},
  year         = {2015},
}