LC/MS (TOF) Analysis of Metanephrine and Normetanephrine in Urine on Ascentis® Express OH5 after Sold Phase Extraction (SPE) using Supel™-Select SCX
Materials
related product
used together
CONDITIONS
sample preparation
SPE (Solid Phase Extraction)
sample/matrix
human urine spiked at 20 ng/mL each analyte
SPE well plate
Supel-Select SCX 96-well plate, 30 mg/well (575664-U)
condition
1 mL 1% formic acid in acetonitrile, 1 mL water
sample addition
1 mL spiked urine
washing
2 mL water, 1 mL methanol:water (25:75)
elution
1 mL 10% ammonium hydroxide in acetonitrile
eluate post-treatment
evaporate and reconstitute in 1 mL acetonitrile
column
Ascentis Express OH5, 5 cm x 2.1 mm I.D., 2.7 μm particles (53749-U)
mobile phase
[A] 5 mM ammonium formate water; [B] 5 mM ammonium formate in acetonitrile; (15:85, A:B)
flow rate
0.3 mL/min
column temp.
35 °C
detector
LC/MS TOF, m/z180.1015, 166.0858
Description
Analysis Note
Metanephrine and normetanephrine are metabolites of epinephrine (adrenaline). Their urinary levels are used in the diagnosis of pheochromocytoma. Here, the analysis of metanephrine and normetanephrine in human urine samples was conducted using Supel-Select SCX to extract the compounds from urine followed by HILIC analysis using Ascentis Express OH5. Analytes were detected in MS/TOF by monitoring their water-loss ions. High purity solvents provided clean, robust operation. Cerilliant CRMs provided reliable quantification.
Legal Information
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany
Supel is a trademark of Sigma-Aldrich Co. LLC