Metanephrines in Urine/Combined analysis - HPLC

Metanephrines in Urine/Combined analysis - HPLC

Order no.: 2020/COMBI
100 Tests
  • 3-Methoxytyramine
  • Metanephrine
  • Normetanephrine

Short analysis time < 10 min
Only 1 HPLC column and 1 Mobile Phase for 2020/COMBI and 6000/COMBI

Description

Clinical relevance

In addition to catecholamines, their metabolic products such as metanephrine and normetanephrine are also important indicators in the diagnosis of various tumours. They have to be determined in addition to the parameters VMA and HVA in order to prevent the risk of false-positive results.

 

Product advantages

  • Short analysis time of less than 10 minutes
  • No changes to setup are needed due to identical column and mobile phase

 

Chromsystems offers an exceptionally sensitive and reliable combined method for detecting pheochromocytomas with an analysis time of less than 10 minutes in the analysis of metanephrine with catecholamines in urine (6000/COMBI). The highly selective separation column and the optimised mobile phase are used for the analysis of both catecholamines and metanephrines, so no change of the chromatographic system is required. The established specific sample preparation using solid phase extraction guarantees simple and safe handling and reproducible results.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

3-Methoxytyramine, Metanephrine, Normetanephrine

Analysis Time

< 10 min

Limit of quantification

4 µg/l

Linearity

4–5000 μg/l

Recovery

93-95 %

Intraassay

CV = 0.6-2.6 %

Interassay

CV = 1.6-2.7 %

Column temperature

30 °C

Flow rate

1.3 ml/min

Specimen

24 h urine (containing 10 ml 25 % HCl)

Sample Preparation

Acidic Hydrolysis

  • Add 100 µl Internal Standard to 1 ml urine in a hydrolysis tube.
  • Adjust pH value to 0.8–1.0, incubate for 30 min at 90–100 °C, then cool down.

Neutralisation

  • Add 6 ml Neutralisation Buffer to hydrolysed urine.

Extraction

  • Apply the sample on the Sample Clean Up Column, discard effluent.
  • Successively wash with 10 ml HPLC water, then add 5 ml Wash Buffer. Discard effluent each time.

Elution

  • Apply 5 ml Elution Buffer on the Sample Clean Up Column and collect the eluate.
  • Add 100 µl Stabilisation Reagent. Mix eluate thoroughly.

HPLC Analysis

  • Inject 10 µl into the HPLC system.

 

Sample Stability

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

Injection Volume

10 µl

Additional Info

For the combined HPLC analysis of catecholamines and metanephrines any isocratic HPLC system with electrochemical detector can be used.

Potential

+770/+840 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.: 2031/COMBI

Mobile Phase, 10 x 1000 ml


Order no.: 2032/COMBI

Calibration Standard


Order no.: 2023

Internal Standard


Order no.: 2024

Neutralisation Buffer


Order no.: 2025

Wash Buffer


Order no.: 2026

Elution Buffer


Order no.: 2027

Stabilisation Reagent


Order no.: 2029/COMBI

Controls

Controls available separately

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