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...
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Elsevier
2023-01-01
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| Series: | Journal of Mass Spectrometry and Advances in the Clinical Lab |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667145X23000020 |
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| author | M. Kruijt N.P.M. Smit J.J. van Ham C.M. Cobbaert L.R. Ruhaak |
| author_facet | M. Kruijt N.P.M. Smit J.J. van Ham C.M. Cobbaert L.R. Ruhaak |
| author_sort | M. Kruijt |
| collection | DOAJ |
| 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, K2-EDTA and K2-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 K2-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. |
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| issn | 2667-145X |
| language | English |
| last_indexed | 2024-04-10T18:29:12Z |
| publishDate | 2023-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Mass Spectrometry and Advances in the Clinical Lab |
| spelling | doaj.art-c099e63cef6346d593289ab494ce4abc2023-02-02T04:50:52ZengElsevierJournal of Mass Spectrometry and Advances in the Clinical Lab2667-145X2023-01-01276165Effects of sample matrix in the measurement of antithrombin by LC-MS: A role for immunocaptureM. Kruijt0N.P.M. Smit1J.J. van Ham2C.M. Cobbaert3L.R. Ruhaak4Department of Clinical Chemistry and Laboratory Medicine, Leiden University Medical Center, Leiden, The Netherlands; Corresponding author at: Leiden University Medical Center, Department of Clinical Chemistry and Laboratory Medicine, Office E2-27, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.Department of Clinical Chemistry and Laboratory Medicine, Leiden University Medical Center, Leiden, The NetherlandsBiobank Facility, Leiden University Medical Center, Leiden, The NetherlandsDepartment of Clinical Chemistry and Laboratory Medicine, Leiden University Medical Center, Leiden, The NetherlandsDepartment of Clinical Chemistry and Laboratory Medicine, Leiden University Medical Center, Leiden, The NetherlandsIntroduction: 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, K2-EDTA and K2-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 K2-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.http://www.sciencedirect.com/science/article/pii/S2667145X23000020Sample matrixQuantitative clinical chemistry proteomicsAnticoagulantsAntithrombinImmunocapture |
| spellingShingle | M. Kruijt N.P.M. Smit J.J. van Ham C.M. Cobbaert L.R. Ruhaak Effects of sample matrix in the measurement of antithrombin by LC-MS: A role for immunocapture Journal of Mass Spectrometry and Advances in the Clinical Lab Sample matrix Quantitative clinical chemistry proteomics Anticoagulants Antithrombin 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 | Sample matrix Quantitative clinical chemistry proteomics Anticoagulants Antithrombin Immunocapture |
| url | http://www.sciencedirect.com/science/article/pii/S2667145X23000020 |
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