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LC/MS (TOF) Analysis of Metanephrine and Normetanephrine in Urine on Ascentis® Express OH5 after Sold Phase Extraction (SPE) using Supel™-Select SCX

LC/MS (TOF) Analysis of Metanephrine and Normetanephrine in Urine on Ascentis® Express OH5 after Sold Phase Extraction (SPE) using Supel™-Select SCX

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)
column temp. 35 C
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
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.
Categories Analytical Chromatography, SPE Applications, Catecholamines
Featured Industry Clinical
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC, Supel is a trademark of Sigma-Aldrich Co. LLC
suitability application for LC-MS

Materials

     
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