Sample Prep Set Total Metanephrines (free + conjugated) with 96 SPE Well Plate

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

• Pipette 200 μl sample/calibrator/MassCheck^® control into each well of the Hydrolysis Plate.
• Add 50 μl Internal Standard Mix + 25 μl Hydrolysis Buffer.
• Seal Hydrolysis Plate with pierceable adhesive seal and cover.
• Hydrolyse in the oven for 45 min at 120 °C.
• Cool for at least 10 min at +2 to +8 °C.
• Place a 96 SPE Well Plate onto a Collection Plate.
• Pipette 50 μl hydrolysate into each well of the 96 SPE Well Plate.
• 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.
• Place 96 SPE Well Plate onto a new Collection Plate.
• Pipette 500 μl Elution Buffer into each well and centrifuge 2 min at 100 x g.
• Pipette 500 μl Dilution Buffer into each well of the Collection Plate.
• Seal with adhesive seal.
• Shake for 2 min at 900 rpm, transfer to autosampler.
• Inject up to 20 μl into the LC-MS/MS system.