Targeted ultra-high performance liquid chromatography-tandem mass spectrometry assay for the quantification of medium-chain phosphatidylcholines in platelets of coronary artery disease patients
In a recent untargeted clinical lipidomics study of platelets of coronary artery disease (CAD) patients, medium-chain phosphatidylcholines (MCPCs) with C8 and C10 fatty acyl residues were found significantly upregulated in the patient group with acute coronary syndrome (ACS) as compared to chronic c...
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Elsevier
2024-11-01
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| Series: | Journal of Chromatography Open |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2772391724000471 |
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| author | Tamara Janker Adrian Brun Adrian Sievers-Engler Kristina Dittrich Meinrad Gawaz Michael Lämmerhofer |
| author_facet | Tamara Janker Adrian Brun Adrian Sievers-Engler Kristina Dittrich Meinrad Gawaz Michael Lämmerhofer |
| author_sort | Tamara Janker |
| collection | DOAJ |
| description | In a recent untargeted clinical lipidomics study of platelets of coronary artery disease (CAD) patients, medium-chain phosphatidylcholines (MCPCs) with C8 and C10 fatty acyl residues were found significantly upregulated in the patient group with acute coronary syndrome (ACS) as compared to chronic coronary syndrome (CCS) and healthy controls. To support this finding, this work presents the development and optimization of a targeted UHPLC-QTrap-MS/MS method with multiple reaction monitoring acquisition for the quantitative analysis of MCPCs (PC 10:0/8:0, PC 16:0/8:0, PC 10:0/20:4 and PC 10:0/10:0) in platelets for biomarker validation. A systematic optimization of chromatographic and mass spectrometric parameters was performed. A charged surface hybrid CSH C18 (1.7 µm, 130 Å) column and fine-tuned gradient elution with 2-propanol/acetonitrile and ammonium acetate as additive to the mobile phase was employed in the final method in ESI negative mode. Four selected PC standards (PC 6:0/6:0, PC 8:0/8:0, PC 10:0/10:0 and PC 12:0/12:0), which cover well the carbon and retention rime range of the target analytes, were used for the optimization process and calibration. Quantification was based on matrix-matched calibration with these four selected commercially available MCPC standards as surrogate calibrants and PC 6:0/6:0(d22) as internal standard. Furthermore, an organic solvent and fatty acyl carbon number-corrected response factor approach gave also accuracies within acceptance limits of bioanalytical validation guidelines and has more generic applicability. Compared to the previous untargeted RPLC-ESI-QTOF-MS/MS method, the optimized targeted UHPLC-QTrap-MS/MS assay showed increased sensitivity and selectivity for the detection of medium-chain PCs in platelet samples of CAD (LOQs in the range of 0.5–5 nmol/L). The method performance parameters indicated its suitability for a future biomarker validation study of MCPCs in platelets. |
| format | Article |
| id | doaj-art-7712b110674344cc8d6cc146eb3e16ee |
| institution | OA Journals |
| issn | 2772-3917 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Chromatography Open |
| spelling | doaj-art-7712b110674344cc8d6cc146eb3e16ee2025-08-20T01:56:34ZengElsevierJournal of Chromatography Open2772-39172024-11-01610016010.1016/j.jcoa.2024.100160Targeted ultra-high performance liquid chromatography-tandem mass spectrometry assay for the quantification of medium-chain phosphatidylcholines in platelets of coronary artery disease patientsTamara Janker0Adrian Brun1Adrian Sievers-Engler2Kristina Dittrich3Meinrad Gawaz4Michael Lämmerhofer5Pharmaceutical (Bio-)Analysis, Institute of Pharmaceutical Sciences, University of Tuebingen, Auf der Morgenstelle 8, 72076 Tuebingen, GermanyPharmaceutical (Bio-)Analysis, Institute of Pharmaceutical Sciences, University of Tuebingen, Auf der Morgenstelle 8, 72076 Tuebingen, GermanyPharmaceutical (Bio-)Analysis, Institute of Pharmaceutical Sciences, University of Tuebingen, Auf der Morgenstelle 8, 72076 Tuebingen, Germany; Corresponding authors. Pharmaceutical (Bio-)Analysis, Institute of Pharmaceutical Sciences, University of Tuebingen, Auf der Morgenstelle 8 72076 Tuebingen, Germany.Pharmaceutical (Bio-)Analysis, Institute of Pharmaceutical Sciences, University of Tuebingen, Auf der Morgenstelle 8, 72076 Tuebingen, GermanyDepartment of Cardiology and Angiology, University Hospital Tuebingen, Eberhard Karls University Tuebingen, Otfried-Mueller-Straße 10 72076 Tuebingen, GermanyPharmaceutical (Bio-)Analysis, Institute of Pharmaceutical Sciences, University of Tuebingen, Auf der Morgenstelle 8, 72076 Tuebingen, Germany; Corresponding authors. Pharmaceutical (Bio-)Analysis, Institute of Pharmaceutical Sciences, University of Tuebingen, Auf der Morgenstelle 8 72076 Tuebingen, Germany.In a recent untargeted clinical lipidomics study of platelets of coronary artery disease (CAD) patients, medium-chain phosphatidylcholines (MCPCs) with C8 and C10 fatty acyl residues were found significantly upregulated in the patient group with acute coronary syndrome (ACS) as compared to chronic coronary syndrome (CCS) and healthy controls. To support this finding, this work presents the development and optimization of a targeted UHPLC-QTrap-MS/MS method with multiple reaction monitoring acquisition for the quantitative analysis of MCPCs (PC 10:0/8:0, PC 16:0/8:0, PC 10:0/20:4 and PC 10:0/10:0) in platelets for biomarker validation. A systematic optimization of chromatographic and mass spectrometric parameters was performed. A charged surface hybrid CSH C18 (1.7 µm, 130 Å) column and fine-tuned gradient elution with 2-propanol/acetonitrile and ammonium acetate as additive to the mobile phase was employed in the final method in ESI negative mode. Four selected PC standards (PC 6:0/6:0, PC 8:0/8:0, PC 10:0/10:0 and PC 12:0/12:0), which cover well the carbon and retention rime range of the target analytes, were used for the optimization process and calibration. Quantification was based on matrix-matched calibration with these four selected commercially available MCPC standards as surrogate calibrants and PC 6:0/6:0(d22) as internal standard. Furthermore, an organic solvent and fatty acyl carbon number-corrected response factor approach gave also accuracies within acceptance limits of bioanalytical validation guidelines and has more generic applicability. Compared to the previous untargeted RPLC-ESI-QTOF-MS/MS method, the optimized targeted UHPLC-QTrap-MS/MS assay showed increased sensitivity and selectivity for the detection of medium-chain PCs in platelet samples of CAD (LOQs in the range of 0.5–5 nmol/L). The method performance parameters indicated its suitability for a future biomarker validation study of MCPCs in platelets.http://www.sciencedirect.com/science/article/pii/S2772391724000471BiomarkerClinical lipidomicsMass spectrometryPhospholipidsAcute coronary syndrome |
| spellingShingle | Tamara Janker Adrian Brun Adrian Sievers-Engler Kristina Dittrich Meinrad Gawaz Michael Lämmerhofer Targeted ultra-high performance liquid chromatography-tandem mass spectrometry assay for the quantification of medium-chain phosphatidylcholines in platelets of coronary artery disease patients Journal of Chromatography Open Biomarker Clinical lipidomics Mass spectrometry Phospholipids Acute coronary syndrome |
| title | Targeted ultra-high performance liquid chromatography-tandem mass spectrometry assay for the quantification of medium-chain phosphatidylcholines in platelets of coronary artery disease patients |
| title_full | Targeted ultra-high performance liquid chromatography-tandem mass spectrometry assay for the quantification of medium-chain phosphatidylcholines in platelets of coronary artery disease patients |
| title_fullStr | Targeted ultra-high performance liquid chromatography-tandem mass spectrometry assay for the quantification of medium-chain phosphatidylcholines in platelets of coronary artery disease patients |
| title_full_unstemmed | Targeted ultra-high performance liquid chromatography-tandem mass spectrometry assay for the quantification of medium-chain phosphatidylcholines in platelets of coronary artery disease patients |
| title_short | Targeted ultra-high performance liquid chromatography-tandem mass spectrometry assay for the quantification of medium-chain phosphatidylcholines in platelets of coronary artery disease patients |
| title_sort | targeted ultra high performance liquid chromatography tandem mass spectrometry assay for the quantification of medium chain phosphatidylcholines in platelets of coronary artery disease patients |
| topic | Biomarker Clinical lipidomics Mass spectrometry Phospholipids Acute coronary syndrome |
| url | http://www.sciencedirect.com/science/article/pii/S2772391724000471 |
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