VMA, HVA, 5-HIAA in Urine - HPLC

VMA, HVA, 5-HIAA in Urine - HPLC

Order no.: 1000/B
100 Tests
  • 5-Hydroxyindoleacetic acid (5-HIAA)
  • Homovanillic acid (HVA)
  • Vanillylmandelic acid (VMA)

No pH-adjustment necessary
Suitable for automated sample preparation
2 Internal standards available

Description

Clinical Relevance

Neuroblastoma is a neoplastic disease that is among the most common malignant cancers occurring during childhood. It is a neuroendocrine tumour resulting in higher excretion of the catecholamine metabolites vanillylmandelic acid (VMA) and homovanillic acid (HVA) in the urine. Therefore verification of these parameters is critical in the diagnosis of neuroblastoma.

In contrast, increased concentrations of hydroxyindoleacetic acid (5-HIAA) detected in the urine are an indication of a carcinoid tumour. The malignant reproduction of enterochromaffin cells in the gastrointestinal tract leads to increased production of the tissue hormone serotonin, whose primary metabolite is 5-HIAA. 

 

Product  advantages

  • Simple and reliable sample preparation
  • Two internal standards if desired
  • Automated sample preparation for Gilson® ASPECTM is available

 

This kit enables routine HPLC determination of VMA, HVA and 5-HIAA in urine. Simple and safe operation guarantees reliable and reproducible results. An additional, optional internal standard (HICA) is also available for especially high precision in the determination of 5-HIAA.

 

Kit content

Mobile Phase, 1000 ml
VMA, HVA, 5-HIAA Calibration Standard (lyoph.), 5 x 1 ml
Internal Standard (iso-VMA), 100 ml
Wash Buffer I, 300 ml
Wash Buffer II, 300 ml
Elution Buffer, 200 ml
Finisher, 10 ml
Sample Clean Up Columns, 50 pcs.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

5-Hydroxyindoleacetic acid (5-HIAA), Homovanillic acid (HVA), Vanillylmandelic acid (VMA)

Analysis Time

< 18 min

Limit of quantification

VMA, HVA, 5-HIAA: 0.5 mg/l

Linearity

VMA: up to 78 mg/l
HVA: up to 51 mg/l
5-HIAA: up to 78 mg/l

Recovery

VMA: 70 %; HVA: 82 %; 5-HIAA: 66 %

Intraassay

CV < 4 %

Interassay

CV < 5 %

Column temperature

ambient (~25 °C)

Flow rate

1 ml/min

Specimen

24 h urine; spontaneous urine (refer the data to urine creatinine)

Pre-analytic Treatment

Determination of VMA/HVA

Collect urine in 10 ml HCl 25 %, pH < 4

 

Determination of 5-HIAA

Collect urine in 10 ml glacial acetic acid, pH 4 - 6

Sample Preparation

  • Extraction: Add 1 ml Internal Standard to 50 µl urine and mix. Apply entire sample to the sample clean up column, draw through by vacuum or centrifugation, discard effluent.
  • Washing: Apply 3 ml Wash Buffer I to the sample clean up column, discard effluent. Apply 2 times 3 ml Wash Buffer II, discard effluents.
  • Elution: Apply 2.0 ml Elution Buffer to the sample clean up column, collect eluate. Add 100 µl Finisher and mix briefly.
  • Inject 10–20 µl into the HPLC system.

Sample Stability

VMA, HVA: at +2 to +8 °C at least 5 days. 5-HIAA: at +2 to +8 °C in the dark max. 3 days. Aliquot and freeze below -18 °C for longer storage.

Injection Volume

10-20 µl

Additional Info

For the HPLC analysis of VMA, HVA and 5-HIAA in urine any isocratic HPLC system with electrochemical detector is suitable.

Potential

approx. +760 mV

Please note

The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.

Kit Components

Components available separately

Mobile Phase


Order no.: 1011

Wash Buffer I


Order no.: 1005

Wash Buffer II


Order no.: 1006

Elution Buffer


Order no.: 1077

Finisher


Order no.: 1013

Calibrators

Calibrators available separately

Controls

Controls available separately