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

Metanephrine: 4.00 µg/l – 20000 µg/l
Normetanephrine: 4.00 µg/l – 20000 µg/l
3-Methoxytyramine: 4.50 µg/l – 10000 µg/l

Different systems might show different performance data.

The information on the sample preparation presented here is abbreviated and is not sufficient for use in the laboratory.
For a detailed step by step description, please refer to the instruction manual.

Hydrolysis in reaction vials
• Pipette 200 µL sample/calibrator/control into a reaction vial.
• Add 50 µL Internal Standard Mix.
• Add 25 µL Hydrolysis Buffer.
• Close the reaction vial, mix briefly.
• Hydrolyse in a water bath for 30 min at 95 °C.
• Cool the reaction vial for at least 10 min.

Sample preparation with Sample Clean Up Columns
• Place a Sample Clean Up Column into a tube.
• Pipette 50 µL of the hydrolysate into the Sample Clean Up Column.
• Add 600 µL Neutralisation Buffer, centrifuge 5 min, discard effluent.
• Add 900 µL Wash Buffer 1, centrifuge 5 min, discard effluent.
• Add 2 x 900 µL Wash Buffer 2, centrifuge each time 1 min, discard effluent.
• Centrifuge 1 min to dryness, discard effluent.
• Place labelled amber glass autosampler vials into clean tubes and then put the Sample Clean Up Columns into the tube with the vial. Thus, the prepared sample is eluted directly into the autosampler vial.
• Pipette 500 µL Elution Buffer into each Sample Clean Up Column and centrifuge 2 min.
• Pipette 500 µL Dilution Buffer into each autosampler vial.
• Close the autosampler vials.
• Mix each vial, transfer to the autosampler.
• Inject up to 20 µL of each eluate into the LC-MS/MS system.

For the sample preparation with 96 SPE well plates, please see Sample Prep Set 80700/96.