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

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 the hydrolysis plate
• Pipette 200 µL sample/calibrator/control into the wells of a Hydrolysis Plate.
• Add 50 µL Internal Standard Mix.
• Add 25 µL Hydrolysis Buffer.
• Seal the Hydrolysis Plate with a Pierceable Adhesive Seal and a Cover.
• Hydrolyse in the oven for 45 min at 120 °C.
• Cool the plate for at least 10 min.

Sample preparation in the 96 SPE well plate
• 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, 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 the 96 SPE Well Plate onto a new Collection Plate.
• Pipette 500 µL Elution Buffer into each well and centrifuge 2 min.
• Pipette 500 µL Dilution Buffer into each well of the Collection Plate.
• Seal the Collection Plate with a Pierceable Adhesive Seal.
• Shake the Collection Plate for 2 min, transfer to the autosampler.
• Inject up to 20 µL of each eluate into the LC-MS/MS system.

For the sample preparation with reaction vials, please see Sample Prep Set 80700/C.