{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T03:15:42Z","timestamp":1792034142496,"version":"4.4.0"},"reference-count":40,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,1,8]],"date-time":"2025-01-08T00:00:00Z","timestamp":1736294400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Genome Canada","award":["265MET"],"award-info":[{"award-number":["265MET"]}]},{"name":"Genome Canada","award":["MC4T"],"award-info":[{"award-number":["MC4T"]}]},{"name":"Genome Canada","award":["42495"],"award-info":[{"award-number":["42495"]}]},{"name":"Canadian Foundation for Innovation","award":["265MET"],"award-info":[{"award-number":["265MET"]}]},{"name":"Canadian Foundation for Innovation","award":["MC4T"],"award-info":[{"award-number":["MC4T"]}]},{"name":"Canadian Foundation for Innovation","award":["42495"],"award-info":[{"award-number":["42495"]}]},{"name":"Segal McGill Chair in Molecular Oncology at McGill University","award":["265MET"],"award-info":[{"award-number":["265MET"]}]},{"name":"Segal McGill Chair in Molecular Oncology at McGill University","award":["MC4T"],"award-info":[{"award-number":["MC4T"]}]},{"name":"Segal McGill Chair in Molecular Oncology at McGill University","award":["42495"],"award-info":[{"award-number":["42495"]}]},{"name":"Warren Y. Soper Charitable Trust","award":["265MET"],"award-info":[{"award-number":["265MET"]}]},{"name":"Warren Y. Soper Charitable Trust","award":["MC4T"],"award-info":[{"award-number":["MC4T"]}]},{"name":"Warren Y. Soper Charitable Trust","award":["42495"],"award-info":[{"award-number":["42495"]}]},{"name":"Alvin Segal Family Foundation","award":["265MET"],"award-info":[{"award-number":["265MET"]}]},{"name":"Alvin Segal Family Foundation","award":["MC4T"],"award-info":[{"award-number":["MC4T"]}]},{"name":"Alvin Segal Family Foundation","award":["42495"],"award-info":[{"award-number":["42495"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>Background: Quantifying urinary catecholamines and metanephrines is essential for the clinical screening and diagnosis of neuroendocrine tumours. HPLC with electrochemical detection (HPLC-ECD) is commonly used for this type of analysis but requires extensive sample cleanup. Simple and rapid dilute-and-shoot LC\u2013multiple-reaction monitoring (MRM)-MS assays have been developed for quantitating these analytes in urine but have not yet been validated according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Methods: A simple dilute-and-shoot sample preparation without derivatization was used. C18 RP-UPLC-MRM-MS and positive-ion ESI were used, usually with two transitions per analyte being monitored. Certified deuterated internal standards were used for each analyte. Results: This assay was validated according to the CLSI C62-A guidelines, including accuracy\/trueness, imprecision, sensitivity, specificity, carryover, stability, and linearity. The final MRM-MS method was compared to the established HPLC-ECD clinical chemistry reference method. The run time was reduced from 25 min to 5 min. Conclusions: A simple, robust, rapid, and cost-effective LC-MRM-MS assay for measuring urinary catecholamines and metanephrines was developed and validated according to the CLSI guidelines. This validated method requires minimal sample manipulation before analysis and provides sensitivity, specificity, and improved precision. The implementation of this assay in clinical laboratories will facilitate early and accurate diagnosis.<\/jats:p>","DOI":"10.3390\/metabo15010030","type":"journal-article","created":{"date-parts":[[2025,1,8]],"date-time":"2025-01-08T04:54:08Z","timestamp":1736312048000},"page":"30","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Multiplexed Dilute-and-Shoot Liquid Chromatography\u2013Multiple-Reaction Monitoring Mass Spectrometry Clinical Assay for Metanephrines and Catecholamines in Human Urine"],"prefix":"10.3390","volume":"15","author":[{"given":"Deema O.","family":"Qasrawi","sequence":"first","affiliation":[{"name":"Segal Cancer Proteomics Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3924-6521","authenticated-orcid":false,"given":"Adriano M. C.","family":"Pimenta","sequence":"additional","affiliation":[{"name":"Segal Cancer Proteomics Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evgeniy V.","family":"Petrotchenko","sequence":"additional","affiliation":[{"name":"Segal Cancer Proteomics Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaun","family":"Eintracht","sequence":"additional","affiliation":[{"name":"Department of Medicine, Sir Mortimer B. Davis Jewish General Hospital, McGill University, Montreal, QC H4A 3J1, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2394-6512","authenticated-orcid":false,"given":"Christoph H.","family":"Borchers","sequence":"additional","affiliation":[{"name":"Segal Cancer Proteomics Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada"},{"name":"Division of Clinical and Translational Research, McGill University, Montreal, QC H4A 3J1, Canada"},{"name":"Gerald Bronfman Department of Oncology, Sir Mortimer B. Davis Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada"},{"name":"Segal Cancer Centre, Lady Davis Institute for Medical Research, Sir Mortimer B. Davis Jewish General Hospital, Montreal, QC H3A 2B4, Canada"},{"name":"Department of Pathology, Sir Mortimer B. Davis Jewish General Hospital, Montreal, QC H3A 2B4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1016\/j.beem.2013.06.004","article-title":"Determination of catecholamines in plasma and urine","volume":"27","author":"Grouzmann","year":"2013","journal-title":"Best Pract. Res. Clin. Endocrinol. Metab."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.clinbiochem.2010.07.006","article-title":"Current status and future developments of LC-MS\/MS in clinical chemistry for quantification of biogenic amines","volume":"44","author":"Kema","year":"2011","journal-title":"Clin. Biochem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1136\/jcp.44.4.269","article-title":"ACP Broadsheet No 127: April 1991. Measurement of catecholamines and their metabolites in urine","volume":"44","author":"Weinkove","year":"1991","journal-title":"J. Clin. Pathol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1915","DOI":"10.1210\/jc.2014-1498","article-title":"Pheochromocytoma and paraganglioma: An endocrine society clinical practice guideline","volume":"99","author":"Lenders","year":"2014","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1258\/000456304322664663","article-title":"Measurement of catecholamines and their metabolites","volume":"41","author":"Peaston","year":"2004","journal-title":"Ann. Clin. Biochem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1016\/j.cca.2019.05.024","article-title":"Liquid chromatography tandem mass spectrometry for plasma metadrenalines","volume":"495","author":"Davies","year":"2019","journal-title":"Clin. Chim. Acta"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1016\/S0076-6879(83)03034-7","article-title":"Microradioenzymic assays for the measurement of catecholamines and serotonin","volume":"103","author":"Cooper","year":"1983","journal-title":"Methods Enzymol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0165-0270(94)90048-5","article-title":"Interference by pH and Ca2+ ions during measurements of catecholamine release in slices of rat amygdala with fast-scan cyclic voltammetry","volume":"52","author":"Jones","year":"1994","journal-title":"J. Neurosci. Methods"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1367","DOI":"10.1039\/AN9921701367","article-title":"Determination of unconjugated catecholamine in urine as dopamine by thermal lens spectrometry","volume":"117","year":"1992","journal-title":"Analyst"},{"key":"ref_10","first-page":"E4","article-title":"Methods of plasma catecholamine measurement including radioimmunoassay","volume":"247","author":"Raum","year":"1984","journal-title":"Am. J. Physiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/j.cca.2021.06.005","article-title":"Measuring steroids from dried blood spots using tandem mass spectrometry to diagnose congenital adrenal hyperplasia","volume":"520","author":"Qasrawi","year":"2021","journal-title":"Clin. Chim. Acta"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1016\/S0039-9140(98)00242-2","article-title":"Advantages and limitations of thermal lens spectrometry over conventional spectrophotometry for absorbance measurements","volume":"48","author":"Georges","year":"1999","journal-title":"Talanta"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.jchemneu.2015.04.002","article-title":"Real-time monitoring of electrically evoked catecholamine signals in the songbird striatum using in vivo fast-scan cyclic voltammetry","volume":"66\u201367","author":"Smith","year":"2015","journal-title":"J. Chem. Neuroanat."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0009-8981(81)90293-X","article-title":"Analysis of urinary metanephrines by reversed-phase high-performance liquid chromatography and electrochemical detection","volume":"110","author":"Krstulovic","year":"1981","journal-title":"Clin. Chim. Acta"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1427","DOI":"10.1001\/jama.287.11.1427","article-title":"Biochemical Diagnosis of Pheochromocytoma: Which Test Is Best?","volume":"287","author":"Lenders","year":"2002","journal-title":"J. Am. Med. Assoc."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1258\/0004563041201626","article-title":"Paracetamol-associated interference in an HPLC-ECD assay for urinary free metadrenalines and catecholamines","volume":"41","author":"Davidson","year":"2004","journal-title":"Ann. Clin. Biochem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"546","DOI":"10.1016\/j.cca.2010.01.012","article-title":"Performance of plasma free metanephrines measured by liquid chromatography-tandem mass spectrometry in the diagnosis of pheochromocytoma","volume":"411","author":"Peaston","year":"2010","journal-title":"Clin. Chim. Acta"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1093\/clinchem\/48.2.323","article-title":"Analysis of Catecholamines in Urine by Positive-Ion Electrospray Tandem Mass Spectrometry","volume":"48","author":"Shushan","year":"2002","journal-title":"Clin. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"7191","DOI":"10.1007\/s00216-017-0681-3","article-title":"Development and validation of a simple, rapid and sensitive LC-MS\/MS method for the measurement of urinary neurotransmitters and their metabolites","volume":"409","author":"Yan","year":"2017","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1309\/AJCPW0TA8OBBNGCK","article-title":"Current and future applications of mass spectrometry to the clinical laboratory","volume":"136","author":"Strathmann","year":"2011","journal-title":"Am. J. Clin. Pathol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.clinbiochem.2020.01.005","article-title":"Very high rate of false positive biochemical results when screening for pheochromocytoma in a large, undifferentiated population with variable indications for testing","volume":"77","author":"Kline","year":"2020","journal-title":"Clin. Biochem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.aca.2018.01.021","article-title":"A sensitive, high-throughput LC-MS\/MS method for measuring catecholamines in low volume serum","volume":"1037","author":"Zheng","year":"2018","journal-title":"Anal. Chim. Acta"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"9072","DOI":"10.1021\/acs.analchem.0c01263","article-title":"In Matrix Derivatization Combined with LC-MS\/MS Results in Ultrasensitive Quantification of Plasma Free Metanephrines and Catecholamines","volume":"92","author":"Bischoff","year":"2020","journal-title":"Anal. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1949","DOI":"10.4155\/bio.11.170","article-title":"Quantitative determination of free and total dopamine in human plasma by LC-MS\/MS: The importance of sample preparation","volume":"3","author":"Hendriks","year":"2011","journal-title":"Bioanalysis"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1016\/j.jchromb.2012.03.026","article-title":"Ultra sensitive measurement of endogenous epinephrine and norepinephrine in human plasma by semi-automated SPE-LC\u2013MS\/MS","volume":"895\u2013896","author":"Zhang","year":"2012","journal-title":"J. Chromatogr. B"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Greer, B., Chevallier, O., Quinn, B., Botana, L.M., and Elliott, C.T. (2021). Redefining dilute and shoot: The evolution of the technique and its application in the analysis of foods and biological matrices by liquid chromatography mass spectrometry. TrAC Trends Anal. Chem., 141.","DOI":"10.1016\/j.trac.2021.116284"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.trac.2013.10.012","article-title":"Dilute-and-shoot-liquid chromatography-mass spectrometry for urine analysis in doping control and analytical toxicology","volume":"55","author":"Deventer","year":"2014","journal-title":"TrAC Trends Anal. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.jchromb.2018.09.012","article-title":"Comprehensive, robust, and sensitive UPLC-MS\/MS analysis of free biogenic monoamines and their metabolites in urine","volume":"1099","author":"Xie","year":"2018","journal-title":"J. Chromatogr. B Anal. Technol. Biomed. Life Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/j.cca.2017.03.004","article-title":"Simple dilute-and-shoot method for urinary vanillylmandelic acid and homovanillic acid by liquid chromatography tandem mass spectrometry","volume":"468","author":"Clark","year":"2017","journal-title":"Clin. Chim. Acta"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1016\/S0731-7085(99)00308-8","article-title":"Evaluation of degradation of urinary catecholamines and metanephrines and deconjugation of their sulfoconjugates using stability-indicating reversed-phase ion-pair HPLC with electrochemical detection","volume":"22","author":"Chan","year":"2000","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1373\/clinchem.2015.238626","article-title":"CLSI C62-A: A New Standard for Clinical Mass Spectrometry","volume":"62","author":"Lynch","year":"2016","journal-title":"Clin. Chem."},{"key":"ref_32","unstructured":"Wayne, P. (2014). CLSI. Liquid Chromatography-Mass Spectrometry Methods, Clinical and Laboratory Standards Institute. Approved Guideline. CLSI C62-A."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1093\/clinchem\/48.3.533","article-title":"Validation of Liquid Chromatography\u2013Tandem Mass Spectrometry Method for Analysis of Urinary Conjugated Metanephrine and Normetanephrine for Screening of Pheochromocytoma","volume":"48","author":"Taylor","year":"2002","journal-title":"Clin. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1373\/clinchem.2003.024703","article-title":"Measurement of plasma free metanephrine and normetanephrine by liquid chromatography-tandem mass spectrometry for diagnosis of pheochromocytoma","volume":"50","author":"Lagerstedt","year":"2004","journal-title":"Clin. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.trac.2016.01.027","article-title":"Impact of LC-MS\/MS on the laboratory diagnosis of catecholamine-producing tumors","volume":"84","author":"Eisenhofer","year":"2016","journal-title":"TrAC Trends Anal. Chem."},{"key":"ref_36","unstructured":"Clinical and Laboratory Standards Institute (2013). Approved Guideline\u2014Second Edition EP9-A3\u2014Measurement Procedure Comparison and Bias Estimation Using Patient Samples, Clinical and Laboratory Standards Institute."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/S0140-6736(86)90837-8","article-title":"Statistical methods for assessing agreement between two methods of clinical measurement","volume":"1","author":"Bland","year":"1986","journal-title":"Lancet"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1959","DOI":"10.1002\/1097-0231(20001115)14:21<1959::AID-RCM117>3.0.CO;2-T","article-title":"High-performance liquid chromatography\/atmospheric pressure chemical ionization mass spectrometric method for the analysis of catecholamines and metanephrines in human urine","volume":"14","author":"Chan","year":"2000","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2068","DOI":"10.1210\/jc.2004-2025","article-title":"Biochemical and clinical manifestations of dopamine-producing paragangliomas: Utility of plasma methoxytyramine","volume":"90","author":"Eisenhofer","year":"2005","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1258\/acb.2012.012112","article-title":"Analysis of plasma 3-methoxytyramine, normetanephrine and metanephrine by ultraperformance liquid chromatography-tandem mass spectrometry: Utility for diagnosis of dopamine-producing metastatic phaeochromocytoma","volume":"50","author":"Peitzsch","year":"2013","journal-title":"Ann. Clin. Biochem."}],"container-title":["Metabolites"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-1989\/15\/1\/30\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:24:50Z","timestamp":1759919090000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-1989\/15\/1\/30"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,8]]},"references-count":40,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,1]]}},"alternative-id":["metabo15010030"],"URL":"https:\/\/doi.org\/10.3390\/metabo15010030","relation":{},"ISSN":["2218-1989"],"issn-type":[{"value":"2218-1989","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,8]]}}}