Preservation of urine free catecholamines and their free O-methylated metabolites with citric acid as an alternative to hydrochloric acid for LC-MS/MS-based analyses
-
Mirko Peitzsch
,
Daniela Pelzel
Abstract
Background: Measurements of urinary fractionated metadrenalines provide a useful screening test to diagnose phaeochromocytoma. Stability of these compounds and their parent catecholamines during and after urine collection is crucial to ensure accuracy of the measurements. Stabilisation with hydrochloric acid (HCl) can promote deconjugation of sulphate-conjugated metadrenalines, indicating a need for alternative preservatives.
Methods: Urine samples with an intrinsically acidic or alkaline pH (5.5–6.9 or 7.1–8.7, respectively) were used to assess stability of free catecholamines and their free O-methylated metabolites over 7 days of room temperature storage. Stabilisation with HCl was compared with ethylenediaminetetraacetic acid/metabisulphite and monobasic citric acid. Catecholamines and metabolites were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Results: Free catecholamines and their O-methylated metabolites were stable in acidic urine samples over 7 days of room temperature storage, independent of the presence or absence of any stabilisation method. In contrast, free catecholamines, but not the free O-methylated metabolites, showed rapid degradation within 24 h and continuing degradation over 7 days in urine samples with an alkaline pH. Adjustment of alkaline urine samples to a pH of 3–5 with HCl or 4.8–5.4 with citric acid completely blocked degradation of catecholamines. Ethylenediaminetetraacetic acid/metabisulphite, although reducing the extent of degradation of catecholamines in alkaline urine, was largely ineffectual as a stabiliser.
Conclusions: Citric acid is equally effective as HCl for stabilisation of urinary free catecholamines and minimises hazards associated with use of strong inorganic acids while avoiding deconjugation of sulphate-conjugated metabolites during simultaneous LC-MS/MS measurements of free catecholamines and their free O-methylated metabolites.
Acknowledgements
Thanks are extended to the DFG for financial support under the grants for KFO 252 “Microenvironment of the Adrenal in Health and Disease” as well as grants EI855/1-1 and EI515/6-1. Further, we extend our thanks to Mrs. Denise Kaden for technical assistance.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organisation(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
References
1. Lenders JW, Duh QY, Eisenhofer G, Gimenez-Roqueplo AP, Grebe SK, Murad MH, et al. Pheochromocytoma and paraganglioma: an endocrine society clinical practice guideline. J Clin Endocrinol Metab 2014;99:1915–42.10.1210/jc.2014-1498Suche in Google Scholar PubMed
2. Peitzsch M, Pelzel D, Glockner S, Prejbisz A, Fassnacht M, Beuschlein F, et al. Simultaneous liquid chromatography tandem mass spectrometric determination of urinary free metanephrines and catecholamines, with comparisons of free and deconjugated metabolites. Clin Chim Acta 2013;418:50–8.10.1016/j.cca.2012.12.031Suche in Google Scholar PubMed
3. Whiting MJ. Simultaneous measurement of urinary metanephrines and catecholamines by liquid chromatography with tandem mass spectrometric detection. Ann Clin Biochem 2009;46:129–36.10.1258/acb.2008.008180Suche in Google Scholar PubMed
4. Davidson DF. Phaeochromocytoma with normal urinary catecholamines: the potential value of urinary free metadrenalines. Ann Clin Biochem 2002;39:557–66.10.1177/000456320203900603Suche in Google Scholar PubMed
5. Boyle JG, Davidson DF, Perry CG, Connell JM. Comparison of diagnostic accuracy of urinary free metanephrines, vanillyl mandelic acid, and catecholamines and plasma catecholamines for diagnosis of pheochromocytoma. J Clin Endocrinol Metab 2007;92:4602–8.10.1210/jc.2005-2668Suche in Google Scholar PubMed
6. Davidson DF, Bradshaw N, Perry CG, Lindsay R, Freel EM. Urinary free (unconjugated) metadrenalines in different hereditary forms of catecholamine-secreting phaeochromocytoma/paraganglioma. Ann Clin Biochem 2012;49:486–90.10.1258/acb.2012.012021Suche in Google Scholar PubMed
7. Eisenhofer G, Peitzsch M. Laboratory evaluation of pheochromocytoma and paraganglioma. Clin Chem 2014;60:1486–99.10.1373/clinchem.2014.224832Suche in Google Scholar PubMed
8. Boomsma F, Alberts G, Vaneijk L, Manintveld AJ, Schalekamp MA. Optimal collection and storage-conditions for catecholamine measurements in human plasma and urine. Clin Chem 1993;39:2503–8.10.1093/clinchem/39.12.2503Suche in Google Scholar
9. Miki K, Sudo A. Effect of urine pH, storage time, and temperature on stability of catecholamines, cortisol, and creatinine. Clin Chem 1998;44:1759–62.10.1093/clinchem/44.8.1759Suche in Google Scholar
10. Peaston RT, Weinkove C. Measurement of catecholamines and their metabolites. Ann Clin Biochem 2004;41:17–38.10.1258/000456304322664663Suche in Google Scholar PubMed
11. Willemsen JJ, Ross HA, Lenders JW, Sweep FC. Stability of urinary fractionated metanephrines and catecholamines during collection, shipment, and storage of samples. Clin Chem 2007;53:268–72.10.1373/clinchem.2006.075218Suche in Google Scholar
12. Roberts NB, Higgins G, Sargazi M. A study on the stability of urinary free catecholamines and free methyl-derivatives at different pH, temperature and time of storage. Clin Chem Lab Med 2010;48:81–7.10.1515/CCLM.2010.017Suche in Google Scholar
13. Simonin J, Gerber-Lemaire S, Centeno C, Seghezzi C, Iglesias K, Abid K, et al. Synthetic calibrators for the analysis of total metanephrines in urine: revisiting the conditions of hydrolysis. Clin Chim Acta 2012;413:998–1003.10.1016/j.cca.2012.02.023Suche in Google Scholar
14. Moleman P. Preservation of urine samples for assay of catecholamines and their metabolites. Clin Chem 1985;31:653–54.10.1093/clinchem/31.4.653aSuche in Google Scholar
15. Lu YP, Ho CS, Mak TW. Formate buffer as preservative for urinary free catecholamine measurement. Ann Clin Biochem 2004;41:39–42.10.1258/000456304322664672Suche in Google Scholar
16. Lee ZS, Critchley JA. Simultaneous measurement of catecholamines and kallikrein in urine using boric acid preservative. Clin Chim Acta 1998;276:89–102.10.1016/S0009-8981(98)00105-3Suche in Google Scholar
17. Gouarne C, Foury A, Duclos M. Critical study of common conditions of storage of glucocorticoids and catecholamines in 24-h urine collected during resting and exercising conditions. Clin Chim Acta 2004;348:207–14.10.1016/j.cccn.2004.05.023Suche in Google Scholar PubMed
Supplemental Material
The online version of this article (DOI: 10.1515/cclm-2015-0044) offers supplementary material, available to authorized users.
©2016 by De Gruyter
Artikel in diesem Heft
- Frontmatter
- Editorial
- Building a bridge to safe diagnosis in health care. The role of the clinical laboratory
- Capillary electrophoresis for the screening and diagnosis of inherited hemoglobin disorders. Ready for prime time?
- Reviews
- Hyperuricemia as risk factor for coronary heart disease incidence and mortality in the general population: a systematic review and meta-analysis
- Towards biomarker-based tests that can facilitate decisions about prevention and management of preeclampsia in low-resource settings
- Mini Review
- Clinical relevance of sST2 in cardiac diseases
- General Clinical Chemistry and Laboratory Medicine
- Preservation of urine free catecholamines and their free O-methylated metabolites with citric acid as an alternative to hydrochloric acid for LC-MS/MS-based analyses
- Performance characteristics of the VIDAS® 25-OH Vitamin D Total assay – comparison with four immunoassays and two liquid chromatography-tandem mass spectrometry methods in a multicentric study
- Differentiation of acute pyelonephritis from other febrile states in children using urinary neutrophil gelatinase-associated lipocalin (uNGAL)
- Early postoperative C-terminal agrin fragment (CAF) serum levels predict graft loss and proteinuria in renal transplant recipients
- Homocitrulline: a new marker for differentiating acute from chronic renal failure
- Hematology and Coagulation
- Development of hemoglobin typing control materials for laboratory investigation of thalassemia and hemoglobinopathies
- Comparison of capillary electrophoresis and high performance liquid chromatography for detection and quantification of hemoglobin New York
- Prevalence of hemostatic alterations in patients with recurrent spontaneous subconjunctival hemorrhage
- Reference Values and Biological Variations
- Reference intervals for 24 laboratory parameters determined in 24-hour urine collections
- First trimester PAPP-A2, PAPP-A and hCGβ in small-for-gestational-age pregnancies
- Cancer Diagnostics
- Clinical utility of one versus two faecal immunochemical test samples in the detection of advanced colorectal neoplasia in symptomatic patients
- Cardiovascular Diseases
- Time from symptom onset influences high-sensitivity troponin T diagnostic accuracy for the diagnosis of acute myocardial infarction
- Prognostic significance of serum albumin level changes in acute ischemic stroke: the role of biological and analytical variation
- Correlates of serum hepcidin levels and its association with cardiovascular disease in an elderly general population
- Infectious Diseases
- Usefulness of several biomarkers in the management of septic patients: C-reactive protein, procalcitonin, presepsin and mid-regional pro-adrenomedullin
- Diabetes
- Performance of strip-based glucose meters and cassette-based blood gas analyzer for monitoring glucose levels in a surgical intensive care setting
- Accuracy and precision assessment of a new blood glucose monitoring system
- Acknowledgment
- Acknowledgment
- Letter to the Editors
- Glypican-1 as a highly sensitive and specific pancreatic cancer biomarker
- Platelet microRNAs are not modulated by systemic heparin in acute coronary syndromes
- Concomitant appearance of two false positive peaks with a true monoclonal one in a patient with plasma cell myeloma
- Anti-streptavidin interferences in Roche thyroid immunoassays: a case report
- Evaluation of the new Methotrexate CMIA assay on the Architect i2000SR
- Liposomal interference on Sysmex XN-series body fluid mode
- Development of an immunomagnetic beads-based test and its application in influenza surveillance
Artikel in diesem Heft
- Frontmatter
- Editorial
- Building a bridge to safe diagnosis in health care. The role of the clinical laboratory
- Capillary electrophoresis for the screening and diagnosis of inherited hemoglobin disorders. Ready for prime time?
- Reviews
- Hyperuricemia as risk factor for coronary heart disease incidence and mortality in the general population: a systematic review and meta-analysis
- Towards biomarker-based tests that can facilitate decisions about prevention and management of preeclampsia in low-resource settings
- Mini Review
- Clinical relevance of sST2 in cardiac diseases
- General Clinical Chemistry and Laboratory Medicine
- Preservation of urine free catecholamines and their free O-methylated metabolites with citric acid as an alternative to hydrochloric acid for LC-MS/MS-based analyses
- Performance characteristics of the VIDAS® 25-OH Vitamin D Total assay – comparison with four immunoassays and two liquid chromatography-tandem mass spectrometry methods in a multicentric study
- Differentiation of acute pyelonephritis from other febrile states in children using urinary neutrophil gelatinase-associated lipocalin (uNGAL)
- Early postoperative C-terminal agrin fragment (CAF) serum levels predict graft loss and proteinuria in renal transplant recipients
- Homocitrulline: a new marker for differentiating acute from chronic renal failure
- Hematology and Coagulation
- Development of hemoglobin typing control materials for laboratory investigation of thalassemia and hemoglobinopathies
- Comparison of capillary electrophoresis and high performance liquid chromatography for detection and quantification of hemoglobin New York
- Prevalence of hemostatic alterations in patients with recurrent spontaneous subconjunctival hemorrhage
- Reference Values and Biological Variations
- Reference intervals for 24 laboratory parameters determined in 24-hour urine collections
- First trimester PAPP-A2, PAPP-A and hCGβ in small-for-gestational-age pregnancies
- Cancer Diagnostics
- Clinical utility of one versus two faecal immunochemical test samples in the detection of advanced colorectal neoplasia in symptomatic patients
- Cardiovascular Diseases
- Time from symptom onset influences high-sensitivity troponin T diagnostic accuracy for the diagnosis of acute myocardial infarction
- Prognostic significance of serum albumin level changes in acute ischemic stroke: the role of biological and analytical variation
- Correlates of serum hepcidin levels and its association with cardiovascular disease in an elderly general population
- Infectious Diseases
- Usefulness of several biomarkers in the management of septic patients: C-reactive protein, procalcitonin, presepsin and mid-regional pro-adrenomedullin
- Diabetes
- Performance of strip-based glucose meters and cassette-based blood gas analyzer for monitoring glucose levels in a surgical intensive care setting
- Accuracy and precision assessment of a new blood glucose monitoring system
- Acknowledgment
- Acknowledgment
- Letter to the Editors
- Glypican-1 as a highly sensitive and specific pancreatic cancer biomarker
- Platelet microRNAs are not modulated by systemic heparin in acute coronary syndromes
- Concomitant appearance of two false positive peaks with a true monoclonal one in a patient with plasma cell myeloma
- Anti-streptavidin interferences in Roche thyroid immunoassays: a case report
- Evaluation of the new Methotrexate CMIA assay on the Architect i2000SR
- Liposomal interference on Sysmex XN-series body fluid mode
- Development of an immunomagnetic beads-based test and its application in influenza surveillance
