6000 HPLC Kit Catecholamines Urine

Catecholamines in Urine - HPLC

Order no.: 6000
100 Tests
  • Adrenaline (Epinephrine)
  • Dopamine
  • Noradrenaline

Easy and safe handling
High resolution
Neutralisation Buffer with integrated indicator for easy pH adjustment
Combined analysis of urinary metanephrines possible

Description

Clinical relevance

The catecholamines adrenaline (epinephrine), noradrenaline (norepinephrine) and dopamine are biogenic amines, which play a central role in the body as hormones and neurotransmitters. Adrenaline and noradrenaline are produced in the adrenal medulla and the sympathetic nervous system. Dopamine is a neurotransmitter in the brain and acts as a local messenger substance in the peripheral nervous system.

The determination of catecholamines has clinical significance in the diagnosis of pheochromocytomas and other tumours affecting the nervous system. These diseases result in the production of surplus catecholamines, which are then excreted in the urine. 24-hour urine collection is the preferred method for tumour screening. The plasma concentration is determined to localise the tumour or to perform pharmacological function tests. Furthermore, the concentrations of adrenaline and noradrenaline are important indicators for cardiac insufficiency, cardiac diseases and arteriosclerosis.

 

Product advantages

  • Simple and safe handling
  • Low detection limits
  • Low analysis costs
  • Ready-to-use components

 

Our kit allows the standard routine analysis of adrenaline, noradrenaline and dopamine in urine with an isocratic HPLC system and electrochemical detector. A very specific sample preparation using solid phase extraction guarantees simple and safe handling of the samples as well as reproducible results.

The kits 2020/COMBI and 6000/COMBI are available as alternative methods that allow the combined determination of catecholamines and metanephrine with one column.

For the HPLC analysis of catecholamines in plasma, there is another method (# 5000) available.

 

Kit content

Mobile Phase, 1000 ml
Catecholamines Calibration Standard, 10 ml
Internal Standard, 10 ml
Neutralisation Buffer, 300 ml
Elution Buffer, 300 ml
Sample Clean Up Columns, 100 pcs.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

Adrenaline (Epinephrine), Dopamine, Noradrenaline

Analysis Time

< 15 min

Limit of quantification

1.6 - 3,6 µg/l

Linearity

up to 500 µg/l

Recovery

68-80 % (dopamine: 91 %)

Intraassay

CV < 3 %

Interassay

CV < 4.1 %

Column temperature

ambient (~25 °C)

Flow rate

0.8-1.3 ml/min

Specimen

24 h urine (containing 10 ml 25 % HCl) or spontaneous urine (refer the data to urine creatinine).

Sample Preparation

  • Mix 3 ml urine with 100 µl Internal Standard and 6 ml Neutralisation Buffer.
  • Add 2 N NaOH until the colour changes from yellow to green or green-grey (see figure).

 

  • Resuspend sample clean up column and allow buffer to flow out.
  • Apply entire urine dilution to the column, discard effluent.
  • Fill up 2 x to the funnel rim with distilled water, allow running through completely. Discard effluent.
  • Apply 6 ml Elution Buffer to the column. Collect eluate.
  • Acidify the eluate with 180 µl 5 N HCl (30 µl 5 N HCl per ml eluate).
  • Inject 20 µl into the HPLC system.

Sample Stability

At +2 to +8 °C at least 5 days. Aliquot and freeze for longer storage.

Injection Volume

20 µl

Additional Info

For the HPLC analysis of catecholamines any isocratic HPLC system with electrochemical detector is suitable.

Potential

approx. 400-500 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