Journal article
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.
- Grouzmann E Laboratoire des Catecholamines et Peptides, Service de Pharmacologie Clinique, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland.
- Centeno C Laboratoire des Catecholamines et Peptides, Service de Pharmacologie Clinique, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland.
- Eugster PJ Laboratoire des Catecholamines et Peptides, Service de Pharmacologie Clinique, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland.
- 2018-05-01
Published in:
- Clinical chemistry and laboratory medicine. - 2018
5-hydroxyindoleacetic acid
UHPLC-MS/MS
carcinoid tumor
dilute-and-shoot
neuroblastoma
urine
vanillylmandelic acid
Biomarkers
Calibration
Carcinoid Tumor
Chromatography, High Pressure Liquid
Homovanillic Acid
Humans
Hydroxyindoleacetic Acid
Limit of Detection
Neuroblastoma
Tandem Mass Spectrometry
Vanilmandelic Acid
English
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.
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.
- Language
-
- English
- Open access status
- closed
- Identifiers
-
- DOI 10.1515/cclm-2017-1120
- PMID 29708876
- Persistent URL
- https://sonar.ch/global/documents/106407
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