Development and validation of a highly-sensitive, quantitative LC-MS/MS assay to evaluate plasma oxytocin
Introduction: Oxytocin is a 9-amino acid peptide that serves as neuromodulator in the human central nervous system. This peptide is implicated in the regulation of diverse behaviors and plays a significant role in positive social interaction. Currently, oxytocin levels are measured using immunoassay...
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Elsevier
2025-04-01
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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/S2667145X25000057 |
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| author | E. Grifnée A. Mackowiak J. Demeuse M. Schoumacher L. Huyghebaert W. Determe T. Dubrowski P. Massonnet S. Peeters G. Scantamburlo E. Cavalier C.Le Goff |
| author_facet | E. Grifnée A. Mackowiak J. Demeuse M. Schoumacher L. Huyghebaert W. Determe T. Dubrowski P. Massonnet S. Peeters G. Scantamburlo E. Cavalier C.Le Goff |
| author_sort | E. Grifnée |
| collection | DOAJ |
| description | Introduction: Oxytocin is a 9-amino acid peptide that serves as neuromodulator in the human central nervous system. This peptide is implicated in the regulation of diverse behaviors and plays a significant role in positive social interaction. Currently, oxytocin levels are measured using immunoassays. However, these methods have several limitations that can lead to false results and erroneous interpretation. Given the remarkably low endogenous level of oxytocin in human plasma (low ng/L levels), we developed and rigorously validated a novel and highly sensitive LC-MS/MS method for oxytocin quantification in plasma. Methods: Oxytocin was initially extracted using solid-phase extraction with an Oasis HLB 30 mg plate and then subjected to LC-MS/MS analysis. PBS-0.1 % BSA served as surrogate matrix for the preparation of validation samples and the calibration curve, ensuring no endogenous interference. The validation design followed the Clinical Laboratory Standards Institute guidelines. Precision, accuracy, and measurement uncertainty were determined using single-nested analysis of variance and e.noval software. Results: A lower limit of quantification of 1 ng/L was achieved. The method was validated for oxytocin concentrations ranging from 1 ng/L to 75 ng/L, with precision (coefficient of variation) below 10 %, accuracy ranging from 94 % to 108 %, and measurement uncertainty below 15 %. Conclusion: In this work, we developed and validated a highly sensitive LC-MS/MS method for the quantification of oxytocin in plasma. Our novel methodology is well-suited for clinical applications. |
| format | Article |
| id | doaj-art-8b35935f0ecf431fbe3a08fcf4ba5576 |
| institution | OA Journals |
| issn | 2667-145X |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Mass Spectrometry and Advances in the Clinical Lab |
| spelling | doaj-art-8b35935f0ecf431fbe3a08fcf4ba55762025-08-20T02:28:19ZengElsevierJournal of Mass Spectrometry and Advances in the Clinical Lab2667-145X2025-04-0136192810.1016/j.jmsacl.2025.02.002Development and validation of a highly-sensitive, quantitative LC-MS/MS assay to evaluate plasma oxytocinE. Grifnée0A. Mackowiak1J. Demeuse2M. Schoumacher3L. Huyghebaert4W. Determe5T. Dubrowski6P. Massonnet7S. Peeters8G. Scantamburlo9E. Cavalier10C.Le Goff11Department of Clinical Chemistry, University Hospital of Liège, Belgium; Corresponding author.Department of Clinical Chemistry, University Hospital of Liège, BelgiumDepartment of Clinical Chemistry, CIRM, University of Liège, BelgiumDepartment of Clinical Chemistry, CIRM, University of Liège, BelgiumDepartment of Clinical Chemistry, University Hospital of Liège, BelgiumDepartment of Clinical Chemistry, CIRM, University of Liège, BelgiumDepartment of Clinical Chemistry, University Hospital of Liège, BelgiumDepartment of Clinical Chemistry, University Hospital of Liège, BelgiumDepartment of Clinical Chemistry, University Hospital of Liège, BelgiumDepartment of Psychiatry, University Hospital of Liège, BelgiumDepartment of Clinical Chemistry, University Hospital of Liège, Belgium; Department of Clinical Chemistry, CIRM, University of Liège, BelgiumDepartment of Clinical Chemistry, University Hospital of Liège, Belgium; Department of Clinical Chemistry, CIRM, University of Liège, BelgiumIntroduction: Oxytocin is a 9-amino acid peptide that serves as neuromodulator in the human central nervous system. This peptide is implicated in the regulation of diverse behaviors and plays a significant role in positive social interaction. Currently, oxytocin levels are measured using immunoassays. However, these methods have several limitations that can lead to false results and erroneous interpretation. Given the remarkably low endogenous level of oxytocin in human plasma (low ng/L levels), we developed and rigorously validated a novel and highly sensitive LC-MS/MS method for oxytocin quantification in plasma. Methods: Oxytocin was initially extracted using solid-phase extraction with an Oasis HLB 30 mg plate and then subjected to LC-MS/MS analysis. PBS-0.1 % BSA served as surrogate matrix for the preparation of validation samples and the calibration curve, ensuring no endogenous interference. The validation design followed the Clinical Laboratory Standards Institute guidelines. Precision, accuracy, and measurement uncertainty were determined using single-nested analysis of variance and e.noval software. Results: A lower limit of quantification of 1 ng/L was achieved. The method was validated for oxytocin concentrations ranging from 1 ng/L to 75 ng/L, with precision (coefficient of variation) below 10 %, accuracy ranging from 94 % to 108 %, and measurement uncertainty below 15 %. Conclusion: In this work, we developed and validated a highly sensitive LC-MS/MS method for the quantification of oxytocin in plasma. Our novel methodology is well-suited for clinical applications.http://www.sciencedirect.com/science/article/pii/S2667145X25000057OxytocinSolid phase extractionLiquid chromatographyTandem mass spectrometryCLSI guidelinesPsychiatric disorders |
| spellingShingle | E. Grifnée A. Mackowiak J. Demeuse M. Schoumacher L. Huyghebaert W. Determe T. Dubrowski P. Massonnet S. Peeters G. Scantamburlo E. Cavalier C.Le Goff Development and validation of a highly-sensitive, quantitative LC-MS/MS assay to evaluate plasma oxytocin Journal of Mass Spectrometry and Advances in the Clinical Lab Oxytocin Solid phase extraction Liquid chromatography Tandem mass spectrometry CLSI guidelines Psychiatric disorders |
| title | Development and validation of a highly-sensitive, quantitative LC-MS/MS assay to evaluate plasma oxytocin |
| title_full | Development and validation of a highly-sensitive, quantitative LC-MS/MS assay to evaluate plasma oxytocin |
| title_fullStr | Development and validation of a highly-sensitive, quantitative LC-MS/MS assay to evaluate plasma oxytocin |
| title_full_unstemmed | Development and validation of a highly-sensitive, quantitative LC-MS/MS assay to evaluate plasma oxytocin |
| title_short | Development and validation of a highly-sensitive, quantitative LC-MS/MS assay to evaluate plasma oxytocin |
| title_sort | development and validation of a highly sensitive quantitative lc ms ms assay to evaluate plasma oxytocin |
| topic | Oxytocin Solid phase extraction Liquid chromatography Tandem mass spectrometry CLSI guidelines Psychiatric disorders |
| url | http://www.sciencedirect.com/science/article/pii/S2667145X25000057 |
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