Mercedes Casado1, Cristina Sierra1, Marta Batllori1, Rafael Artuch2,3,4, Aida Ormazabal1,5. 1. Clinical Biochemistry Department, Institut de Recerca Sant Joan de Déu, Barcelona, Spain. 2. Clinical Biochemistry Department, Institut de Recerca Sant Joan de Déu, Barcelona, Spain. rartuch@hsjdbcn.org. 3. CIBERER-Instituto de Salud Carlos III, Barcelona, Spain. rartuch@hsjdbcn.org. 4. Clinical Biochemistry Department, Hospital Sant Joan de Déu, Barcelona, Spain. rartuch@hsjdbcn.org. 5. CIBERER-Instituto de Salud Carlos III, Barcelona, Spain.
Abstract
INTRODUCTION: Amino acid analysis in biological fluids is essential for the study of inborn errors of metabolism (IEM) and other diseases. OBJECTIVES: Our aim was to develop a UPLC-MS/MS procedure for the analysis of 25 amino acids and identification of 17 related compounds. METHODS: Sample treatment conditions were optimized for plasma, urine, cerebrospinal fluid (CSF) and dried blood spots. Amino acids and related compounds were analyzed on a Waters ACQUITY UPLC H-class instrument with a reversed-phase C-18 column using water and acetonitrile with 0.1% formic acid as the mobile phases (run time = 9 min). The detection was performed with a Waters Xevo TQD triple-quadrupole mass spectrometer using positive electrospray ionization in the multiple reaction monitoring mode. RESULTS: The method linearity, intra-assay and inter-assay precision, detection limit, quantification limit and trueness analysis displayed adequate results in both physiological and pathological conditions. Method comparison was performed between UPLC-MS/MS and ion exchange chromatography (IEC) with ninhydrin derivatization, and the methods showed good agreement, except for 4-hydroxyproline, aspartate and citrulline. Paediatrics age-related reference values in plasma, urine and CSF were established and patients with different IEM were easily identified. CONCLUSION: We report a modified UPLC-MS/MS procedure for the analysis of 42 amino acids and related compounds in different specimens. The method is fast, sensitive and robust, and it has been validated to be an alternative to the traditional IEC procedure as the routine method used in metabolic laboratories. The method greatly decreases the run time of the analysis while displaying good metrological results.
INTRODUCTION: Amino acid analysis in biological fluids is essential for the study of inborn errors of metabolism (IEM) and other diseases. OBJECTIVES: Our aim was to develop a UPLC-MS/MS procedure for the analysis of 25 amino acids and identification of 17 related compounds. METHODS: Sample treatment conditions were optimized for plasma, urine, cerebrospinal fluid (CSF) and dried blood spots. Amino acids and related compounds were analyzed on a Waters ACQUITY UPLC H-class instrument with a reversed-phase C-18 column using water and acetonitrile with 0.1% formic acid as the mobile phases (run time = 9 min). The detection was performed with a Waters Xevo TQD triple-quadrupole mass spectrometer using positive electrospray ionization in the multiple reaction monitoring mode. RESULTS: The method linearity, intra-assay and inter-assay precision, detection limit, quantification limit and trueness analysis displayed adequate results in both physiological and pathological conditions. Method comparison was performed between UPLC-MS/MS and ion exchange chromatography (IEC) with ninhydrin derivatization, and the methods showed good agreement, except for 4-hydroxyproline, aspartate and citrulline. Paediatrics age-related reference values in plasma, urine and CSF were established and patients with different IEM were easily identified. CONCLUSION: We report a modified UPLC-MS/MS procedure for the analysis of 42 amino acids and related compounds in different specimens. The method is fast, sensitive and robust, and it has been validated to be an alternative to the traditional IEC procedure as the routine method used in metabolic laboratories. The method greatly decreases the run time of the analysis while displaying good metrological results.
Entities:
Keywords:
Amino acids; Inborn errors of metabolism; Tandem mass spectrometry; UPLC
Authors: Barbara Plecko; Karl Paul; Eduard Paschke; Sylvia Stoeckler-Ipsiroglu; Eduard Struys; Cornelis Jakobs; Hans Hartmann; Thomas Luecke; Matteo di Capua; Christoph Korenke; Christiane Hikel; Elke Reutershahn; Michael Freilinger; Fritz Baumeister; Friedrich Bosch; Wolfgang Erwa Journal: Hum Mutat Date: 2007-01 Impact factor: 4.878
Authors: P Krumpochova; B Bruyneel; D Molenaar; A Koukou; M Wuhrer; W M A Niessen; M Giera Journal: J Pharm Biomed Anal Date: 2015-06-05 Impact factor: 3.935
Authors: Hubertus C M T Prinsen; B G M Schiebergen-Bronkhorst; M W Roeleveld; J J M Jans; M G M de Sain-van der Velden; G Visser; P M van Hasselt; N M Verhoeven-Duif Journal: J Inherit Metab Dis Date: 2016-04-21 Impact factor: 4.982
Authors: Yuan Zhang; M Rebecca Glineburg; Venkatesha Basrur; Kevin Conlon; Shannon E Wright; Amy Krans; Deborah A Hall; Peter K Todd Journal: Hum Mol Genet Date: 2022-07-21 Impact factor: 5.121
Authors: Pau B Esparza-Moltó; Inés Romero-Carramiñana; Cristina Núñez de Arenas; Marta P Pereira; Noelia Blanco; Beatriz Pardo; Georgina R Bates; Carla Sánchez-Castillo; Rafael Artuch; Michael P Murphy; José A Esteban; José M Cuezva Journal: PLoS Biol Date: 2021-05-13 Impact factor: 8.029
Authors: Marta Batllori; Mercedes Casado; Cristina Sierra; Maria Del Carmen Salgado; Laura Marti-Sanchez; Joan Maynou; Guerau Fernandez; Angels Garcia-Cazorla; Aida Ormazabal; Marta Molero-Luis; Rafael Artuch Journal: Fluids Barriers CNS Date: 2019-11-14
Authors: Tessa M A Peters; Udo F H Engelke; Siebolt de Boer; Ed van der Heeft; Cynthia Pritsch; Purva Kulkarni; Ron A Wevers; Michèl A A P Willemsen; Marcel M Verbeek; Karlien L M Coene Journal: J Inherit Metab Dis Date: 2020-05-23 Impact factor: 4.982
Authors: Cristina Jou; Andres Nascimento; Anna Codina; Julio Montoya; Ester López-Gallardo; Sonia Emperador; Eduardo Ruiz-Pesini; Raquel Montero; Daniel Natera-de Benito; Carlos I Ortez; Jesus Marquez; Maria V Zelaya; Alfonso Gutierrez-Mata; Carmen Badosa; Laura Carrera-García; Jesica Expósito-Escudero; Monica Roldán; Yolanda Camara; Ramon Marti; Isidre Ferrer; Cecilia Jimenez-Mallebrera; Rafael Artuch Journal: Int J Mol Sci Date: 2022-09-20 Impact factor: 6.208