VMA, HVA, 5-HIAA in Urine - HPLC

Order No.: 1000/B, for 100 tests
Parameters:
5-Hydroxyindoleacetic Acid (5-HIAA), Homovanillic Acid (HVA), Vanillylmandelic Acid (VMA)

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

5-Hydroxyindoleacetic Acid (5-HIAA)

Homovanillic Acid (HVA)

Vanillylmandelic Acid(VMA)

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.

More Information
Number of Tests 100
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 %
Analysis Time < 18 min
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.
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

Injection Volume 10-20 µl
Flow rate 1 ml/min
Column temperature ambient (~25 °C)
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.
Method of Analysis HPLC
Parameter 5-Hydroxyindoleacetic Acid (5-HIAA), Homovanillic Acid (HVA), Vanillylmandelic Acid (VMA)
Complete Specification
Linearity: up to 75 mg/l (VMA)
up to 50 mg/l (HVA)
up to 75 mg/l (5-HIAA)
Limit of quantification: 0.5 mg/l (VMA, HVA, 5-HIAA)
Intra-assay: CV ~ 4 %
Inter-assay: CV ~ 6 %
Recovery: 85 % (VMA)
83 % (HVA)
106 % (5-HIAA)
Run time: aproxx. 20 min

HPLC parameters

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

Injection volume: 10-20 µl
Flow rate: 0.8-1.3 ml/min
Potential: aproxx. +700 to +850 mV
Column temperature: ambient (~25 °C)
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
  1. 1011 HPLC mobile phase VMA HVA 5-HIAA urine
    Mobile Phase
    Order no.: 1011
    VMA, HVA and 5-HIAA in Urine - HPLC
    Details
  2. 1011 HPLC mobile phase VMA HVA 5-HIAA urine
    Mobile Phase, 10 x 1000 ml
    Order no.: 1012
    VMA, HVA and 5-HIAA in Urine - HPLC
    Details
  3. 1003-B HPLC urine calibration standard VMA HVA 5-HIAA
    Calibration Standard
    Order no.: 1003/B
    VMA, HVA, 5-HIAA in Urine - HPLC
    Details
  4. 1004-B HPLC internal standard urine
    Internal Standard (iso-VMA)
    Order no.: 1004/B
    VMA, HVA and 5-HIAA in Urine - HPLC
    Details
  5. 1005 HPLC wash buffer 1 VMA HVA 5-HIAA urine
    Wash Buffer I
    Order no.: 1005
    VMA, HVA and 5-HIAA in Urine - HPLC
    Details
  6. 1006 HPLC wash buffer 2 VMA HVA 5-HIAA urine
    Wash Buffer II
    Order no.: 1006
    VMA, HVA and 5-HIAA in Urine - HPLC
    Details
  7. 1077 HPLC elution buffer VMA HVA 5-HIAA urine
    Elution Buffer
    Order no.: 1077
    VMA, HVA and 5-HIAA in Urine - HPLC
    Details
  8. 1013 HPLC finisher VMA HVA 5-HIAA urine
    Finisher
    Order no.: 1013
    VMA, HVA and 5-HIAA in Urine - HPLC
    Details
  9. 1008 HPLC sample clean up columns VMA HVA 5-HIAA urine
    Sample Clean Up Columns
    Order no.: 1008
    VMA, HVA and 5-HIAA in Urine - HPLC
    Details
  1. 1009 HPLC VMA HVA 5-HIAA urine calibration standard
    VMA, HVA, 5-HIAA Urine Calibration Standard
    Order no.: 1009
    VMA, HVA, 5-HIAA in Urine - HPLC
    Details
  2. 1003-B HPLC urine calibration standard VMA HVA 5-HIAA
    Calibration Standard
    Order no.: 1003/B
    VMA, HVA, 5-HIAA in Urine - HPLC
    Details
  1. 0040 0050 HPLC endocrine urine controls
    Endocrine Urine Controls
    Order no.: 0040/0050
    Details
  1. 1100-B HPLC column VMA HVA 5-HIAA urine
    HPLC column
    Order no.: 1100/B
    VMA, HVA and 5-HIAA in Urine
    Details
  2. 1099 HPLC Potential Optimisation Mix (VMA, iso-VMA)
    Potential Optimisation Mix (VMA, iso-VMA)
    Order no.: 1099
    VMA, HVA and 5-HIAA in Urine - HPLC
    Details
  3. 51303-B HPLC internal standard urine HICA 5-HIAA
    Internal Standard (HICA), 5-HIAA
    Order no.: 51303/B
    VMA, HVA and 5-HIAA in urine - HPLC
    Details
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