5-HIAA in Urine - HPLC

5-HIAA in Urine - HPLC

Order no.: 51000
100 Tests
  • 5-Hydroxyindoleacetic acid (5-HIAA)

Easy sample preparation
No exact pH-adjustment necessary
The alternative to immunoassay methods
Compatible with the VMA/HVA/5-HIAA method

Description

Clinical relevance

A number of diseases are associated with pathological changes in serotonin metabolism. In carcinoid tumour, a malignant hyperplasia of the enterochromaffin cells of the gastrointestinal tract, there is a marked increase in serotonin release into blood plasma. This tissue hormone is metabolised enzymatically in the liver by monoamine oxidase (MAO) and aldehyde dehydrogenase (ADH) to 5-hydroxyindole-3-acetic acid (5-HIAA) and subsequently excreted via the kidney. Endocrine-active carcinoid thus results in an increased concentration of 5-HIAA in the urine. The quantitative determination of 5-HIAA in urine is therefore used for the diagnosis of the carcinoid tumour and for therapy monitoring.

 

Product advantages

  • Easy sample preparation
  • No exact pH adjustment required
  • The alternative to immunoassay methods
  • Compatible with the Chromsystems VMA/HVA/5-HIAA method

 

This kit for the determination of 5-HIAA in urine provides a rapid and reliable sample preparation procedure with only one precipitation step. The kit contains all the components in a ready-to-use form for sample preparation and HPLC analysis. A carefully optimised chromatographic separation is part of the Chromsystems complete solution. A selected HPLC column and a mobile phase optimised specifically for this separation permit a reliable quantification.

Urine samples prepared with this kit can also be analysed with column and mobile phase of the VMA/HVA/5-HIAA Chromsystems kit.

 

Kit content for 100 analyses:

Mobile Phase, 1000 ml
Internal Standard, 10 ml
Urine Calibration Standard (lyoph.), 2 x 5 ml
Precipitation Reagent, 75 ml
Reaction vials, 100 pcs. 

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

5-Hydroxyindoleacetic acid (5-HIAA)

Analysis Time

~ 9 min

Limit of quantification

0.5 mg/l

Linearity

up to at least 25 mg/l

Recovery

101 %

Intraassay

CV = 2.6-6.9 %

Interassay

CV = 4.0-4.1 %

Column temperature

ambient (~25 °C)

Flow rate

0.8 - 1.3 ml/min

Specimen

24 h-urines collected with 10 ml glacial acetic acid.

Pre-analytic Treatment

24h-urine collected with 10 ml glacial acetic acid, pH 4 - 6. Stored in the dark at +2 to +8 °C patient samples are stable up to 3 days. For longer storage periods patient samples should be frozen below -18 °C.

Sample Preparation

Place a 200 μl aliquot of the acidified urine sample in a test tube or reaction vial.
Add 100 μl Internal Standard.
Then add 700 μl Precipitation Reagent.
Mix thoroughly.
Centrifuge in a bench-top centrifuge (2 min with at least 10000 x g).
Dilute 500 μl supernatant with 500 μl ultrapure water (HPLC grade). Mix thoroughly.
Inject 10–20 μl of this solution into the HPLC system.

Sample Stability

Stored at +2 to +8 °C the samples are stable up to 3 days.

Injection Volume

10-20 µl

Additional Info

For the HPLC analysis 5-HIAA in urine any isocratic HPLC system with electrochemical detector is suitable.

Potential

+620 to +680 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

Calibrators

Calibrators available separately

Controls

Controls available separately