Sample Prep Set Total Metanephrines (free + conjugated) with Sample Clean Up Columns

Sample Prep Set Total Metanephrines (free + conjugated) with Sample Clean Up Columns

Order no.: 80700/C
96 Tests
  • 3-Methoxytyramine
  • Metanephrine
  • Normetanephrine

Isotopically labelled internal standard for each analyte
6PLUS1® Multilevel Calibrator Set and MassCheck® controls

Description

MassChrom® Biogenic Amines/Metabolites in Urine – LC-MS/MS
With Sample Clean Up Columns, for manual sample prep procedure

 

Note: In addition to this Sample Prep Set the products of the Chromatographic Platform (Analytical Column, Mobile Phases A and B, Rinsing Solution) are required.

 

Clinical relevance

This Sample Prep Set allows the quantitative determination of total metanephrine, normetanephrine and 3-methoxytyramine in patient urine samples by LC-MS/MS.
One typical application for this kit is the analysis of patient samples with a suspicion of catecholamine-secreting tumours.

 

Product advantages

  • Isotopically labelled internal standard for each analyte
  • 6PLUS1® Multilevel Calibrator Set and MassCheck®controls ensure high precision of results.


This Sample Prep Set encompasses all required components for an efficient and fast sample preparation.

There is also a method using 96 SPE well plates (80700/96) as well as the corresponding fully automated workflow solution available for laboratories with a higher sample throughput.

Chromatogram

Technical Data

Method of Analysis

LC-MS/MS

Number of Tests

96

Parameter

3-Methoxytyramine, Metanephrine, Normetanephrine

Analysis Time

4 min

Linearity

Metanephrine, Normetanephrine up to 20 mg/l
3-Methoxytyramine up to 10 mg/l

Recovery

92–107 %

Intraassay

CV = 2.5–5.6 %

Interassay

CV = 3.2–4.9 %

Specimen

24h urine or spontaneous urine

Pre-analytic Treatment

24 h urine is collected in a suitable container with 10 ml of 10–25 % HCl.

Sample Preparation

  • Pipette 200 μl sample/calibrator/MassCheck® control into a reaction vial.
  • Add 50 μl Internal Standard Mix+ 25 μl Hydrolysis Buffer.
  • Close reaction vial, mix briefly.
  • Hydrolyse for 30 min at 95 °C.
  • Cool for at least 10 min at +2 to +8 °C.
  • Pipette 50 μl of the hydrolysate into a Sample Clean Up Column.
  • Add 600 μl Neutralisation Buffer.
  • Centrifuge 5 min at 150 x g, discard effluent.
  • Add 0.9 ml Wash Buffer 1, centrifuge 5 min at 150 x g, discard effluent.
  • Add 2 x 0.9 ml Wash Buffer 2, centrifuge each time 1 min at 150 x g, discard effluent.
  • Centrifuge 1 min at 2000 x g to dryness, discard effluent.
  • Change collection vials. (Use glass vials, see instruction manual).
  • Pipette 500 μl Elution Buffer into each Sample Clean Up Column and centrifuge 2 min at 100 x g.
  • Pipette 500 μl Dilution Buffer into each glass vial.
  • Close glass vials.
  • Vortex, transfer to autosampler.
  • Inject up to 20 μl into the LC-MS/MS system.

Sample Stability

Samples are stable for at least 7 days at +2°C to +8°C in amber coloured vials. For longer storage periods keep samples frozen below -18°C.

Injection Volume

≤ 20 µl

Ionisation

ESI positive

MS/MS-Mode

MRM

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

Calibrators

Controls

Controls available separately