Catecholamines in Urine - HPLC

Order No.: 6000, for 100 tests
Parameters:
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

Adrenaline (Epinephrine)

Dopamine

Noradrenaline

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.

More Information
Number of Tests 100
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 %

Analysis Time < 15 min
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.
Specimen 24 h urine (containing 10 ml 25 % HCl) or spontaneous urine (refer the data to urine creatinine).
Injection Volume 20 µl
Flow rate 0.8-1.3 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 Adrenaline (Epinephrine), Dopamine, Noradrenaline
Complete Specification
Linearity: up to 1000 µg/l
Limit of quantification: 3 µg/l
Intra-assay: CV < 5 %
Inter-assay: CV < 6 %
Recovery: 69-81 %
Run time: approx. 15 min

HPLC parameters

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

Injection volume: 20 µl
Flow rate: 0.8-1.3 ml/min
Potential: approx. 400-500 mV
Column temperature: ambient (~25 °C)
Additional Info

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

Potential approx. 400-500 mV
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  2. 5001 HPLC mobile phase catecholamines plasma urine
    Mobile Phase, 10 x 1000 ml
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  3. 6003 HPLC urine calibration standard catecholamines
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  4. 6004 HPLC internal standard catecholamines urine
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    Order no.: 6004
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  5. 6055 HPLC neutralisation buffer catecholamines urine
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    Order no.: 6055
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    Details
  6. 6006 HPLC elution buffer catecholamines urine
    Elution Buffer
    Order no.: 6006
    Catecholamines in Urine - HPLC
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  7. 6007 HPLC sample clean up columns catecholamines urine
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  1. 6009 HPLC Calibration Standard Catecholamines Urine
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  2. 6003 HPLC urine calibration standard catecholamines
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  1. 6100 HPLC column catecholamines urine
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