Validation of an LC-MS/MS method for urinary homovanillic and vanillylmandelic ACIDS and application to the diagnosis of neuroblastoma

Background: Urinary catecholamine metabolites are well-established biomarkers for neuroblastoma (NB). Homovanillic acid (HVA) and vanillylmandelic acid (VMA) are the most frequently measured metabolites within SIOPEN – Catecholamine Working Group laboratories. Here, we evaluated the performance of a...

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Main Authors: Lucilla Rossi, Yvette A.H. Matser, Sebastiano Barco, Alessia Cafaro, Federica Pigliasco, Margherita Biondi, Fabrizio Mancin, Maria van der Ham, Monique G.M. de Sain-van der Velden, Shifra Ash, Maja Beck Popovic, André B.P. van Kuilenburg, Massimo Conte, Alberto Garaventa, Godelieve A.M. Tytgat, Giuliana Cangemi
Format: Article
Language:English
Published: Elsevier 2025-04-01
Series:Journal of Mass Spectrometry and Advances in the Clinical Lab
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Online Access:http://www.sciencedirect.com/science/article/pii/S2667145X25000148
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author Lucilla Rossi
Yvette A.H. Matser
Sebastiano Barco
Alessia Cafaro
Federica Pigliasco
Margherita Biondi
Fabrizio Mancin
Maria van der Ham
Monique G.M. de Sain-van der Velden
Shifra Ash
Maja Beck Popovic
André B.P. van Kuilenburg
Massimo Conte
Alberto Garaventa
Godelieve A.M. Tytgat
Giuliana Cangemi
author_facet Lucilla Rossi
Yvette A.H. Matser
Sebastiano Barco
Alessia Cafaro
Federica Pigliasco
Margherita Biondi
Fabrizio Mancin
Maria van der Ham
Monique G.M. de Sain-van der Velden
Shifra Ash
Maja Beck Popovic
André B.P. van Kuilenburg
Massimo Conte
Alberto Garaventa
Godelieve A.M. Tytgat
Giuliana Cangemi
author_sort Lucilla Rossi
collection DOAJ
description Background: Urinary catecholamine metabolites are well-established biomarkers for neuroblastoma (NB). Homovanillic acid (HVA) and vanillylmandelic acid (VMA) are the most frequently measured metabolites within SIOPEN – Catecholamine Working Group laboratories. Here, we evaluated the performance of a new LC-MS/MS in vitro diagnostic (IVD) kit for HVA and VMA to facilitate inter-laboratory harmonization. Methods: HVA and VMA and their deuterated internal standards were analyzed with a commercial method, on a ThermoFisher Quantiva LC-MS/MS. Validation was performed first using internal quality control and external quality assessment (IQC and EQA) samples. Next by clinical validation on 120 samples, previously tested by HPLC-ECD. Finally, 36 samples were exchanged between SIOPEN reference laboratories and analyzed by three methods. Results: Using QCs and EQA the method was validated in a wide calibration range (4.61–830 µmol/L for HVA and 4.44–800 µmol/L for VMA). Intra-day CVs (n = 5) were 7 and 8 % for HVA and 5 and 6 % for VMA for QC low and QC high, respectively; Inter-day CV% were 7 and 3 % for HVA and 2 and 7 % for VMA at QC low and QC high, respectively. Its application to 120 clinical samples confirmed a high diagnostic accuracy. The inter-laboratory quality control assessment showed interchangeable results (p = 0,73 and p = 0.15 for HVA and VMA, respectively). Conclusion: The LC-MS/MS IVD method could be considered a useful tool for clinical laboratories involved in the measurement of catecholamines, contributing to harmonization efforts.
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spelling doaj-art-0847aa65be924aee97ca59d2a2120c972025-08-20T02:56:12ZengElsevierJournal of Mass Spectrometry and Advances in the Clinical Lab2667-145X2025-04-0136738110.1016/j.jmsacl.2025.04.007Validation of an LC-MS/MS method for urinary homovanillic and vanillylmandelic ACIDS and application to the diagnosis of neuroblastomaLucilla Rossi0Yvette A.H. Matser1Sebastiano Barco2Alessia Cafaro3Federica Pigliasco4Margherita Biondi5Fabrizio Mancin6Maria van der Ham7Monique G.M. de Sain-van der Velden8Shifra Ash9Maja Beck Popovic10André B.P. van Kuilenburg11Massimo Conte12Alberto Garaventa13Godelieve A.M. Tytgat14Giuliana Cangemi15Biochemistry, Pharmacology and Newborn Screening Unit, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, Genoa, Italy; Department of Chemical Sciences – DiSC, University of Padua, Padua, ItalyPrincess Máxima Center for Paediatric Oncology, Utrecht, The Netherlands; Laboratory of Genetic Metabolic Diseases, Amsterdam UMC University of Amsterdam, Amsterdam, The NetherlandsBiochemistry, Pharmacology and Newborn Screening Unit, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, Genoa, Italy; Corresponding author at: Chromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, Via Gerolamo Gaslini 5, 16147 Genoa, Italy.Biochemistry, Pharmacology and Newborn Screening Unit, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, Genoa, ItalyBiochemistry, Pharmacology and Newborn Screening Unit, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, Genoa, ItalyBiochemistry, Pharmacology and Newborn Screening Unit, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, Genoa, ItalyDepartment of Chemical Sciences – DiSC, University of Padua, Padua, ItalyDepartment of Genetics, Section Metabolic Diagnostics, Wilhelmina Children’s Hospital, Utrecht, The NetherlandsDepartment of Genetics, Section Metabolic Diagnostics, Wilhelmina Children’s Hospital, Utrecht, The NetherlandsRuth Rappaport Children’s Hospital, Haifa, IsraelUniversity Hospital Lausanne, Lausanne, SwitzerlandLaboratory of Genetic Metabolic Diseases, Amsterdam UMC University of Amsterdam, Amsterdam, The Netherlands; Cancer Center Amsterdam, Imaging and Biomarkers, Amsterdam, The NetherlandsPediatric Oncology and Hematology Department, IRCCS Istituto Giannina Gaslini, Genova, ItalyPediatric Oncology and Hematology Department, IRCCS Istituto Giannina Gaslini, Genova, ItalyPrincess Máxima Center for Paediatric Oncology, Utrecht, The NetherlandsBiochemistry, Pharmacology and Newborn Screening Unit, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, Genoa, ItalyBackground: Urinary catecholamine metabolites are well-established biomarkers for neuroblastoma (NB). Homovanillic acid (HVA) and vanillylmandelic acid (VMA) are the most frequently measured metabolites within SIOPEN – Catecholamine Working Group laboratories. Here, we evaluated the performance of a new LC-MS/MS in vitro diagnostic (IVD) kit for HVA and VMA to facilitate inter-laboratory harmonization. Methods: HVA and VMA and their deuterated internal standards were analyzed with a commercial method, on a ThermoFisher Quantiva LC-MS/MS. Validation was performed first using internal quality control and external quality assessment (IQC and EQA) samples. Next by clinical validation on 120 samples, previously tested by HPLC-ECD. Finally, 36 samples were exchanged between SIOPEN reference laboratories and analyzed by three methods. Results: Using QCs and EQA the method was validated in a wide calibration range (4.61–830 µmol/L for HVA and 4.44–800 µmol/L for VMA). Intra-day CVs (n = 5) were 7 and 8 % for HVA and 5 and 6 % for VMA for QC low and QC high, respectively; Inter-day CV% were 7 and 3 % for HVA and 2 and 7 % for VMA at QC low and QC high, respectively. Its application to 120 clinical samples confirmed a high diagnostic accuracy. The inter-laboratory quality control assessment showed interchangeable results (p = 0,73 and p = 0.15 for HVA and VMA, respectively). Conclusion: The LC-MS/MS IVD method could be considered a useful tool for clinical laboratories involved in the measurement of catecholamines, contributing to harmonization efforts.http://www.sciencedirect.com/science/article/pii/S2667145X25000148NeuroblastomaUrinary catecholamineLiquid Chromatography-tandem Mass SpectrometryPediatricsInter-laboratory harmonizationHomovanillic acid
spellingShingle Lucilla Rossi
Yvette A.H. Matser
Sebastiano Barco
Alessia Cafaro
Federica Pigliasco
Margherita Biondi
Fabrizio Mancin
Maria van der Ham
Monique G.M. de Sain-van der Velden
Shifra Ash
Maja Beck Popovic
André B.P. van Kuilenburg
Massimo Conte
Alberto Garaventa
Godelieve A.M. Tytgat
Giuliana Cangemi
Validation of an LC-MS/MS method for urinary homovanillic and vanillylmandelic ACIDS and application to the diagnosis of neuroblastoma
Journal of Mass Spectrometry and Advances in the Clinical Lab
Neuroblastoma
Urinary catecholamine
Liquid Chromatography-tandem Mass Spectrometry
Pediatrics
Inter-laboratory harmonization
Homovanillic acid
title Validation of an LC-MS/MS method for urinary homovanillic and vanillylmandelic ACIDS and application to the diagnosis of neuroblastoma
title_full Validation of an LC-MS/MS method for urinary homovanillic and vanillylmandelic ACIDS and application to the diagnosis of neuroblastoma
title_fullStr Validation of an LC-MS/MS method for urinary homovanillic and vanillylmandelic ACIDS and application to the diagnosis of neuroblastoma
title_full_unstemmed Validation of an LC-MS/MS method for urinary homovanillic and vanillylmandelic ACIDS and application to the diagnosis of neuroblastoma
title_short Validation of an LC-MS/MS method for urinary homovanillic and vanillylmandelic ACIDS and application to the diagnosis of neuroblastoma
title_sort validation of an lc ms ms method for urinary homovanillic and vanillylmandelic acids and application to the diagnosis of neuroblastoma
topic Neuroblastoma
Urinary catecholamine
Liquid Chromatography-tandem Mass Spectrometry
Pediatrics
Inter-laboratory harmonization
Homovanillic acid
url http://www.sciencedirect.com/science/article/pii/S2667145X25000148
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