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Quantification of vanillylmandelic acid, homovanillic acid and 5-hydroxyindoleacetic acid in urine using a dilute-and-shoot and ultra-high pressure liquid chromatography tandem mass spectrometry method

  • Eric Grouzmann EMAIL logo , Catherine Centeno and Philippe J. Eugster
Published/Copyright: May 1, 2018
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 56 Issue 9

Abstract

Background:

Urinary vanillylmandelic acid (VMA) and homovanillic acid (HVA) are biomarkers for the diagnosis and follow-up of neuroblastoma, whereas urinary 5-hydroxyindoleacetic acid (5-HIAA) is used to assess a carcinoid tumor. These analytes are conventionally analyzed in a single run by chromatography (LC) coupled with electrochemical detection (LC-ECD) using commercial kits. A rapid dilute-and-shoot LC tandem mass spectrometry (LC-MS/MS) assay was validated in order to replace the LC-ECD method and therefore improve analytical specificity and throughput.

Methods:

Sample preparation was carried out by dilution of the urine sample with a solution containing the deuterated internal standards. The separation was achieved on an ultra-high pressure LC system with MS detection using a triple quadrupole mass spectrometer. The method was validated according to the current EMA and FDA guidelines.

Results:

The full chromatographic run was achieved in 8 min. The method validation showed excellent linearity (r2>0.999 for all three analytes), precision (CV <15%), negligible matrix effect (recoveries >90%), low carryover (<1%) and LLOQ of 0.25, 0.4 and 0.4 μM for VMA, HVA and 5-HIAA, respectively. Deming fits and Bland-Altman analyses showed no significant differences between the values obtained between the two assays.

Conclusions:

The LC-MS/MS method proposed in this study is fast and robust, and the simple sample preparation saves time and avoids the additional costs of dedicated kits used for the LC-ECD assays by switching to LC-MS/MS. Additionally, the near-perfect correlation observed herein between both assays allows the previously established reference ranges to be maintained.

Keywords: carcinoid tumor; dilute-and-shoot; 5-hydroxyindoleacetic acid; neuroblastoma; UHPLC-MS/MS; urine; vanillylmandelic acid
Corresponding author: Dr. Eric Grouzmann, Laboratoire des Catecholamines et Peptides, Service de Pharmacologie Clinique, Centre Hospitalier Universitaire Vaudois, CH-1011 Lausanne, Switzerland, Phone: +41213140741, Fax: +41213147835

Acknowledgments

The authors thank Dr. Steve Bruce for the careful reading of the manuscript.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(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.

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Supplementary Material:

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2017-1120).

Received: 2017-12-01
Accepted: 2018-03-27
Published Online: 2018-05-01
Published in Print: 2018-08-28

©2018 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2017-1120
Keywords for this article
carcinoid tumor; dilute-and-shoot; 5-hydroxyindoleacetic acid; neuroblastoma; UHPLC-MS/MS; urine; vanillylmandelic acid

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. Clinical Chemistry and Laboratory Medicine continues to shine brightly in the constellation of laboratory medicine
  4. The Theranos saga and the consequences
  5. Innovative approaches in diabetes diagnosis and monitoring: less invasive, less expensive… but less, equally or more efficient?
  6. Reviews
  7. Exploring the microbiota to better understand gastrointestinal cancers physiology
  8. Linking type 2 diabetes and gynecological cancer: an introductory overview
  9. Mini Reviews
  10. MicroRNAs as predictive biomarkers of response to tyrosine kinase inhibitor therapy in metastatic renal cell carcinoma
  11. Salivary biomarkers and cardiovascular disease: a systematic review
  12. Opinion Paper
  13. The meteoric rise and dramatic fall of Theranos: lessons learned for the diagnostic industry
  14. General Clinical Chemistry and Laboratory Medicine
  15. Uncertainty evaluation in clinical chemistry, immunoassay, hematology and coagulation analytes using only external quality assessment data
  16. Measurement uncertainty and metrological traceability of whole blood cyclosporin A mass concentration results obtained by UHPLC-MS/MS
  17. Computer-assisted interventions in the clinical laboratory process improve the diagnosis and treatment of severe vitamin B12 deficiency
  18. Trueness, precision and stability of the LIAISON 1-84 parathyroid hormone (PTH) third-generation assay: comparison to existing intact PTH assays
  19. Fibroblast growth factor 23 and renal function among young and healthy individuals
  20. Optimizing charge state distribution is a prerequisite for accurate protein biomarker quantification with LC-MS/MS, as illustrated by hepcidin measurement
  21. Quantification of human complement C2 protein using an automated turbidimetric immunoassay
  22. EE score: an index for simple differentiation of homozygous hemoglobin E and hemoglobin E-β0-thalassemia
  23. Reference Values and Biological Variations
  24. Algorithm on age partitioning for estimation of reference intervals using clinical laboratory database exemplified with plasma creatinine
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  26. Cancer Diagnostics
  27. Quantification of vanillylmandelic acid, homovanillic acid and 5-hydroxyindoleacetic acid in urine using a dilute-and-shoot and ultra-high pressure liquid chromatography tandem mass spectrometry method
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