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.

• 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.