Multiplexed Dilute-and-Shoot Liquid Chromatography–Multiple-Reaction Monitoring Mass Spectrometry Clinical Assay for Metanephrines and Catecholamines in Human Urine
<b>Background:</b> 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...
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| author | Deema O. Qasrawi Adriano M. C. Pimenta Evgeniy V. Petrotchenko Shaun Eintracht Christoph H. Borchers |
| author_facet | Deema O. Qasrawi Adriano M. C. Pimenta Evgeniy V. Petrotchenko Shaun Eintracht Christoph H. Borchers |
| author_sort | Deema O. Qasrawi |
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| description | <b>Background:</b> 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–multiple-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. <b>Methods:</b> 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. <b>Results:</b> 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. <b>Conclusions:</b> 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. |
| format | Article |
| id | doaj-art-00139cf58dea45b5832749a7077d6857 |
| institution | Kabale University |
| issn | 2218-1989 |
| language | English |
| publishDate | 2025-01-01 |
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| series | Metabolites |
| spelling | doaj-art-00139cf58dea45b5832749a7077d68572025-01-24T13:41:13ZengMDPI AGMetabolites2218-19892025-01-011513010.3390/metabo15010030Multiplexed Dilute-and-Shoot Liquid Chromatography–Multiple-Reaction Monitoring Mass Spectrometry Clinical Assay for Metanephrines and Catecholamines in Human UrineDeema O. Qasrawi0Adriano M. C. Pimenta1Evgeniy V. Petrotchenko2Shaun Eintracht3Christoph H. Borchers4Segal Cancer Proteomics Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, CanadaSegal Cancer Proteomics Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, CanadaSegal Cancer Proteomics Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, CanadaDepartment of Medicine, Sir Mortimer B. Davis Jewish General Hospital, McGill University, Montreal, QC H4A 3J1, CanadaSegal Cancer Proteomics Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada<b>Background:</b> 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–multiple-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. <b>Methods:</b> 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. <b>Results:</b> 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. <b>Conclusions:</b> 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.https://www.mdpi.com/2218-1989/15/1/30liquid chromatography–multiple-reaction monitoring mass spectrometry (LC-MRM-MS)metanephrinescatecholaminesadrenal glandpheochromocytomaurine |
| spellingShingle | Deema O. Qasrawi Adriano M. C. Pimenta Evgeniy V. Petrotchenko Shaun Eintracht Christoph H. Borchers Multiplexed Dilute-and-Shoot Liquid Chromatography–Multiple-Reaction Monitoring Mass Spectrometry Clinical Assay for Metanephrines and Catecholamines in Human Urine Metabolites liquid chromatography–multiple-reaction monitoring mass spectrometry (LC-MRM-MS) metanephrines catecholamines adrenal gland pheochromocytoma urine |
| title | Multiplexed Dilute-and-Shoot Liquid Chromatography–Multiple-Reaction Monitoring Mass Spectrometry Clinical Assay for Metanephrines and Catecholamines in Human Urine |
| title_full | Multiplexed Dilute-and-Shoot Liquid Chromatography–Multiple-Reaction Monitoring Mass Spectrometry Clinical Assay for Metanephrines and Catecholamines in Human Urine |
| title_fullStr | Multiplexed Dilute-and-Shoot Liquid Chromatography–Multiple-Reaction Monitoring Mass Spectrometry Clinical Assay for Metanephrines and Catecholamines in Human Urine |
| title_full_unstemmed | Multiplexed Dilute-and-Shoot Liquid Chromatography–Multiple-Reaction Monitoring Mass Spectrometry Clinical Assay for Metanephrines and Catecholamines in Human Urine |
| title_short | Multiplexed Dilute-and-Shoot Liquid Chromatography–Multiple-Reaction Monitoring Mass Spectrometry Clinical Assay for Metanephrines and Catecholamines in Human Urine |
| title_sort | multiplexed dilute and shoot liquid chromatography multiple reaction monitoring mass spectrometry clinical assay for metanephrines and catecholamines in human urine |
| topic | liquid chromatography–multiple-reaction monitoring mass spectrometry (LC-MRM-MS) metanephrines catecholamines adrenal gland pheochromocytoma urine |
| url | https://www.mdpi.com/2218-1989/15/1/30 |
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