Catecholamines in Plasma - HPLC

Catecholamines in Plasma - HPLC

Order no.: 5000
200 Tests
  • Adrenaline (Epinephrine)
  • Dopamine
  • Noradrenaline

Easy and safe handling
All components are ready-for-use
High sensitivity
Low analysis costs

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 established kit allows the standard routine analysis of adrenaline, noradrenaline and dopamine in serum/plasma 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.

For the HPLC analysis of catecholamines in urine, there is another method (# 6000) available.

 

Kit content

Mobile Phase, 1000 ml
Extraction Buffer, 100 ml
Catecholamines Calibration Standard, 10 ml
Internal Standard, 10 ml
Wash Buffer, 300 ml
Elution Buffer, 25 ml
Sample Clean Up Columns, 50 pcs.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

200

Parameter

Adrenaline (Epinephrine), Dopamine, Noradrenaline

Analysis Time

< 20 min

Limit of quantification

Adrenaline, Noradrenaline: 18-43 ng/l
Dopamine: 81 ng/l

Linearity

up to 2000 ng/l

Recovery

> 70 % (dopamine > 40 %)

Intraassay

CV < 3.8 %

Interassay

CV < 6 % (dopamine ~13 %)

Column temperature

ambient (~25 °C)

Flow rate

0.8-1.3 ml/min

Specimen

Plasma

Pre-analytic Treatment

Sample drawing

  • Blood samples should be drawn from the recumbent patient.
  • 20 µl stabilisation solutuion (900 mg EGTA, 700 mg glutathione in 0,5 M NaOH ad 8 ml, pH 7-7.5 with H2O ad 10 ml) is required per ml blood.
  • Mix, centrifuge.

Sample Preparation

  • Add 500 µl Extraction Buffer to a sample clean up cartridge, shake briefly.
  • Apply 1 to 1.5 ml plasma and 50 µl Internal Standard.
  • Mix for 10 min by inverting.
  • Remove the bottom plug and remove supernatant by vacuum or centrifugation, discard effluent.
  • 3 times: apply 1 ml Wash Buffer, shake briefly, centrifuge (1 min at 2000 rpm, after the last wash step 2 min at 4000 rpm), discard effluent.
  • Mount the plastic tube to the outlet of the cartridge, add 120 µl Elution Buffer, shake briefly, incubate for 5 min. Vortex 30 s.
  • Centrifuge for 1 min at 2000 rpm into the attached plastic tube. Collect eluat and mix well.
  • Inject 20-50 µl into the HPLC system.

Sample Stability

At ambient temperature up to 2 hours; below -18 °C up to 9 months

Injection Volume

20-50 µl

Additional Info

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

Potential

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

Mobile Phase


Order no.: 5001

Extraction Buffer


Order no.: 5011

Internal Standard


Order no.: 5004

Wash Buffer


Order no.: 5005

Elution Buffer


Order no.: 5006

Controls

Controls available separately

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