Development and validation of a liquid chromatography/tandem mass spectrometry method for the simultaneous quantification of serotonin and thromboxane B2 from activated platelets

Research output: Contribution to journalArticle

Abstract

Introduction: Availability of a rapid and reliable platelet activation assay avoiding limitations of current techniques would be valuable to diagnose heparin-induced thrombocytopenia and platelet secretion disorders. Objectives: The first aim was to develop and validate an ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to quantify in a single run TxB2 synthesized and serotonin released from platelets. The second aim was to use our method in association with light transmission aggregometry (LTA) to select good platelet responders for the diagnosis of HIT. Methods: Electrospray ionization and chromatographic separation were optimized for the simultaneous dosage of serotonin and TxB2. The method was validated according to the European Medicines Agency (EMA) guideline for bioanalytical method validation. LTA was performed with monoclonal anti-CD9 (clone ALB6) as platelet activator to select good responders. Results: Detection was performed using a tandem mass spectrometer with alternated positive and negative electrospray ionization. The total run time was 6 minutes. The method was validated for calibration curves, precision, accuracy, lower limit of quantification, carry-over, selectivity, and matrix effect. Platelet response to ALB6 was highly variable among donors. Conclusion: We developed and validated a UHPLC-MS/MS method for the simultaneous quantification of TxB2 and serotonin.

Original languageEnglish
Pages (from-to)663-671
Number of pages9
JournalInternational Journal of Laboratory Hematology
Volume40
Issue number6
DOIs
Publication statusPublished - 1 Dec 2018

Fingerprint

Thromboxane B2
Liquid chromatography
Platelets
Tandem Mass Spectrometry
Liquid Chromatography
Mass spectrometry
Serotonin
Blood Platelets
Electrospray ionization
Light transmission
Light
Mass spectrometers
Platelet Activation
Medicine
Heparin
Assays
Thrombocytopenia
Calibration
Chemical activation
Association reactions

Keywords

  • Platelet
  • Serotonin
  • Thromboxane B2
  • Triple quadrupole
  • UHPLC-MS/MS
  • thromboxane B2
  • platelet
  • triple quadrupole

Cite this

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title = "Development and validation of a liquid chromatography/tandem mass spectrometry method for the simultaneous quantification of serotonin and thromboxane B2 from activated platelets",
abstract = "Introduction: Availability of a rapid and reliable platelet activation assay avoiding limitations of current techniques would be valuable to diagnose heparin-induced thrombocytopenia and platelet secretion disorders. Objectives: The first aim was to develop and validate an ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to quantify in a single run TxB2 synthesized and serotonin released from platelets. The second aim was to use our method in association with light transmission aggregometry (LTA) to select good platelet responders for the diagnosis of HIT. Methods: Electrospray ionization and chromatographic separation were optimized for the simultaneous dosage of serotonin and TxB2. The method was validated according to the European Medicines Agency (EMA) guideline for bioanalytical method validation. LTA was performed with monoclonal anti-CD9 (clone ALB6) as platelet activator to select good responders. Results: Detection was performed using a tandem mass spectrometer with alternated positive and negative electrospray ionization. The total run time was 6 minutes. The method was validated for calibration curves, precision, accuracy, lower limit of quantification, carry-over, selectivity, and matrix effect. Platelet response to ALB6 was highly variable among donors. Conclusion: We developed and validated a UHPLC-MS/MS method for the simultaneous quantification of TxB2 and serotonin.",
keywords = "Platelet, Serotonin, Thromboxane B2, Triple quadrupole, UHPLC-MS/MS, thromboxane B2, platelet, triple quadrupole",
author = "Valentine Minet and Jonathan Evrard and Christelle Vancraeynest and Dogn{\'e}, {Jean Michel} and Fran{\cc}ois Mullier and Lionel Pochet",
year = "2018",
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doi = "10.1111/ijlh.12901",
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pages = "663--671",
journal = "International Journal of Laboratory Hematology",
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T1 - Development and validation of a liquid chromatography/tandem mass spectrometry method for the simultaneous quantification of serotonin and thromboxane B2 from activated platelets

AU - Minet, Valentine

AU - Evrard, Jonathan

AU - Vancraeynest, Christelle

AU - Dogné, Jean Michel

AU - Mullier, François

AU - Pochet, Lionel

PY - 2018/12/1

Y1 - 2018/12/1

N2 - Introduction: Availability of a rapid and reliable platelet activation assay avoiding limitations of current techniques would be valuable to diagnose heparin-induced thrombocytopenia and platelet secretion disorders. Objectives: The first aim was to develop and validate an ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to quantify in a single run TxB2 synthesized and serotonin released from platelets. The second aim was to use our method in association with light transmission aggregometry (LTA) to select good platelet responders for the diagnosis of HIT. Methods: Electrospray ionization and chromatographic separation were optimized for the simultaneous dosage of serotonin and TxB2. The method was validated according to the European Medicines Agency (EMA) guideline for bioanalytical method validation. LTA was performed with monoclonal anti-CD9 (clone ALB6) as platelet activator to select good responders. Results: Detection was performed using a tandem mass spectrometer with alternated positive and negative electrospray ionization. The total run time was 6 minutes. The method was validated for calibration curves, precision, accuracy, lower limit of quantification, carry-over, selectivity, and matrix effect. Platelet response to ALB6 was highly variable among donors. Conclusion: We developed and validated a UHPLC-MS/MS method for the simultaneous quantification of TxB2 and serotonin.

AB - Introduction: Availability of a rapid and reliable platelet activation assay avoiding limitations of current techniques would be valuable to diagnose heparin-induced thrombocytopenia and platelet secretion disorders. Objectives: The first aim was to develop and validate an ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to quantify in a single run TxB2 synthesized and serotonin released from platelets. The second aim was to use our method in association with light transmission aggregometry (LTA) to select good platelet responders for the diagnosis of HIT. Methods: Electrospray ionization and chromatographic separation were optimized for the simultaneous dosage of serotonin and TxB2. The method was validated according to the European Medicines Agency (EMA) guideline for bioanalytical method validation. LTA was performed with monoclonal anti-CD9 (clone ALB6) as platelet activator to select good responders. Results: Detection was performed using a tandem mass spectrometer with alternated positive and negative electrospray ionization. The total run time was 6 minutes. The method was validated for calibration curves, precision, accuracy, lower limit of quantification, carry-over, selectivity, and matrix effect. Platelet response to ALB6 was highly variable among donors. Conclusion: We developed and validated a UHPLC-MS/MS method for the simultaneous quantification of TxB2 and serotonin.

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