Metanephrines in Urine - HPLC

Metanephrines in Urine - HPLC

Order no.: 2020
100 Tests
  • 3-Methoxytyramine
  • Metanephrine
  • Normetanephrine
No pH adjustment after hydrolysis
Only 1 sample clean up column required
Reliable separation of interferences

Description

Clinical relevance

The biogenic amines normetanephrine, metanephrine, and 3-methoxytyramine are the primary metabolites produced in catecholamine metabolism. They are important parameters in diagnosing certain tumours in the adrenal medulla such as pheochromocytomas, neuroblastomas, ganglioneuromas and melanoblastomas. They have to be determined in addition to adrenaline, noradrenaline, dopamine, VMA and HVA in order to lower the risk of false-positive results.

 

Product advantages

  • Only 1 sample clean up column required
  • Effective removal of interferences
  • Combined catecholamine/metanephrine analysis in urine is possible

 

This kit enables a reliable routine HPLC determination of metanephrine, normetanephrine, and 3-methoxytyramine in urine. The conjugated metanephrines are reliably released using acid hydrolysis. The pH level required for solid phase extraction can be adjusted very easily using the colour change indicator by adding the neutralisation buffer. The chromatographic determination is run on an isocratic HPLC system with an electrochemical detector. Optimised column and mobile phase ensure the reliable separation of possible interfering substances.

 

Kit content

Mobile Phase, 1000 ml
Calibration Standard, 10 ml
Internal Standard, 10 ml
Neutralisation Buffer, 300 ml
Wash Buffer, 250 ml
Elution Buffer, 250 ml
Sample Clean Up Columns, 100 pcs.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

3-Methoxytyramine, Metanephrine, Normetanephrine

Analysis Time

aproxx. 14 min

Limit of quantification

5 µg/l

Linearity

up to 5000 µg/l

Recovery

~ 85 %

Intraassay

CV < 4 %

Interassay

CV < 6 %

Column temperature

ambient (~25 °C)

Flow rate

1.0-1.2 ml/min

Specimen

24 h urine (containing 25 % HCl); spontaneous urine (refer the data to urine creatinine)

Pre-analytic Treatment

Specimens

  • 24 h urine (containing 25 % HCl)
  • Spontaneous urine (refer the data to urine creatinine)

Stability of specimens

  • At +2 to +8 °C at least 5 days

Aliquot and freeze for longer storage.

Sample Preparation

  • Acidic hydrolysis: Pipette 100 µl Internal Standard into a hydrolyis tube, add 1 ml urine, adjust pH with 2 N HCl to 0.8 to 1.0. Incubate in the sealed tube at 90 to 100 °C for 30 min, then cool down immediately.
  • Neutralisation: Add 6 ml Neutralisation Buffer to the hydrolysed urine. If the sample colour does not change to purple, add 2 N NaOH until colour changes.
  • Extraktion: Apply the entire sample to the sample clean up column, discard eluate.
  • Washing: Apply 10 ml distilled water and then 5 ml Wash Buffer, discard eluates.
  • Elution: Apply 5 ml Elution Buffer to the sample clean up column and collect eluate. Mix each eluate briefly.
  • HPLC: Add 30 µl glacial acetic acid per ml eluate. Inject 20 µl of the acidified eluate 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

HPLC parameters

For the Chromsystems HPLC analysis of metanephrines in urine any isocratic HPLC system with electrochemical detector is suitable.

Potential

aproxx. +780 to +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.: 2021

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

Calibrators

Calibrators available separately

Calibration Standard


Order no.: 2023

Controls

Controls available separately