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Effects of sample matrix in the measurement of antithrombin by LC-MS: A role for immunocapture
INTRODUCTION: The sample matrix composition, which is greatly affected by the type of blood collection tube used during phlebotomy, is of major importance in laboratory testing as it can influence test results. We developed an LC-MRM-MS test to molecularly characterize antithrombin in citrate plasma...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9860377/ https://www.ncbi.nlm.nih.gov/pubmed/36691568 http://dx.doi.org/10.1016/j.jmsacl.2023.01.002 |
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author | Kruijt, M. Smit, N.P.M. van Ham, J.J. Cobbaert, C.M. Ruhaak, L.R. |
author_facet | Kruijt, M. Smit, N.P.M. van Ham, J.J. Cobbaert, C.M. Ruhaak, L.R. |
author_sort | Kruijt, M. |
collection | PubMed |
description | INTRODUCTION: The sample matrix composition, which is greatly affected by the type of blood collection tube used during phlebotomy, is of major importance in laboratory testing as it can influence test results. We developed an LC-MRM-MS test to molecularly characterize antithrombin in citrate plasma. The test principle differs greatly from traditional laboratory tests and the influence of varying plasma sample matrices is largely unknown. OBJECTIVES: To identify whether variations in sample matrix affect the LC-MRM-MS test for antithrombin and assess whether sample pre-processing by immunocapture reduces matrix-specific effects. METHODS: Samples (n = 45) originating from four different blood collection tubes (sodium citrate, lithium heparin, K(2)-EDTA and K(2)-EDTA with protease inhibitors) were processed directly or after immunocapture. Antithrombin was digested into proteotypic peptides, which were monitored by LC-MRM-MS. Results from lithium heparin and the K(2)-EDTA matrices were compared to the standard sample matrix, sodium citrate, using Deming regression analysis and repeated measures one-way ANOVA. RESULTS: Deming regression analysis of directly processed samples revealed slopes deviating >5% from the line of identity for at least six out of 22 peptides in all matrices. Significant differences between all matrices were found upon analysis by ANOVA for at least 10 peptides. Pre-processing by immunocapture led to slopes within 5% of the line of identity for nearly all peptides of the matrices. Furthermore, significant differences between matrices after immunocapture were only observed for four peptides. CONCLUSION: Variations in the sample matrix affect the measurement of antithrombin by LC-MRM-MS, but observed effects are greatly reduced upon pre-processing by immunocapture. |
format | Online Article Text |
id | pubmed-9860377 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-98603772023-01-22 Effects of sample matrix in the measurement of antithrombin by LC-MS: A role for immunocapture Kruijt, M. Smit, N.P.M. van Ham, J.J. Cobbaert, C.M. Ruhaak, L.R. J Mass Spectrom Adv Clin Lab Articles from the special issue on Biospecimen Pre-Analytics: Identifying and Controlling Ex Vivo Distortion of In Vivo Reality Edited by Chad Borges, Anne Bendt, Robert Gurke and Julijana Ivanisevic INTRODUCTION: The sample matrix composition, which is greatly affected by the type of blood collection tube used during phlebotomy, is of major importance in laboratory testing as it can influence test results. We developed an LC-MRM-MS test to molecularly characterize antithrombin in citrate plasma. The test principle differs greatly from traditional laboratory tests and the influence of varying plasma sample matrices is largely unknown. OBJECTIVES: To identify whether variations in sample matrix affect the LC-MRM-MS test for antithrombin and assess whether sample pre-processing by immunocapture reduces matrix-specific effects. METHODS: Samples (n = 45) originating from four different blood collection tubes (sodium citrate, lithium heparin, K(2)-EDTA and K(2)-EDTA with protease inhibitors) were processed directly or after immunocapture. Antithrombin was digested into proteotypic peptides, which were monitored by LC-MRM-MS. Results from lithium heparin and the K(2)-EDTA matrices were compared to the standard sample matrix, sodium citrate, using Deming regression analysis and repeated measures one-way ANOVA. RESULTS: Deming regression analysis of directly processed samples revealed slopes deviating >5% from the line of identity for at least six out of 22 peptides in all matrices. Significant differences between all matrices were found upon analysis by ANOVA for at least 10 peptides. Pre-processing by immunocapture led to slopes within 5% of the line of identity for nearly all peptides of the matrices. Furthermore, significant differences between matrices after immunocapture were only observed for four peptides. CONCLUSION: Variations in the sample matrix affect the measurement of antithrombin by LC-MRM-MS, but observed effects are greatly reduced upon pre-processing by immunocapture. Elsevier 2023-01-07 /pmc/articles/PMC9860377/ /pubmed/36691568 http://dx.doi.org/10.1016/j.jmsacl.2023.01.002 Text en © 2023 THE AUTHORS https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Articles from the special issue on Biospecimen Pre-Analytics: Identifying and Controlling Ex Vivo Distortion of In Vivo Reality Edited by Chad Borges, Anne Bendt, Robert Gurke and Julijana Ivanisevic Kruijt, M. Smit, N.P.M. van Ham, J.J. Cobbaert, C.M. Ruhaak, L.R. Effects of sample matrix in the measurement of antithrombin by LC-MS: A role for immunocapture |
title | Effects of sample matrix in the measurement of antithrombin by LC-MS: A role for immunocapture |
title_full | Effects of sample matrix in the measurement of antithrombin by LC-MS: A role for immunocapture |
title_fullStr | Effects of sample matrix in the measurement of antithrombin by LC-MS: A role for immunocapture |
title_full_unstemmed | Effects of sample matrix in the measurement of antithrombin by LC-MS: A role for immunocapture |
title_short | Effects of sample matrix in the measurement of antithrombin by LC-MS: A role for immunocapture |
title_sort | effects of sample matrix in the measurement of antithrombin by lc-ms: a role for immunocapture |
topic | Articles from the special issue on Biospecimen Pre-Analytics: Identifying and Controlling Ex Vivo Distortion of In Vivo Reality Edited by Chad Borges, Anne Bendt, Robert Gurke and Julijana Ivanisevic |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9860377/ https://www.ncbi.nlm.nih.gov/pubmed/36691568 http://dx.doi.org/10.1016/j.jmsacl.2023.01.002 |
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