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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.
