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Catecholamines and Metabolites in Human Urine on Raptor Biphenyl by LC-MS/MS

LC_CF0703
PeakstR (min)Precursor IonProduct Ion
1.Norepinephrine-d61.14158.16111.26
2.Norepinephrine1.14152.07106.91
3.Epinephrine-d61.32172.16111.86
4.Epinephrine1.33166.07135.00
5.Normetanephrine-d31.51169.00136.96
6.Normetanephrine1.52166.00148.99
PeakstR (min)Precursor IonProduct Ion
7.Dopamine-d41.55141.1694.69
8.Dopamine1.56136.9764.66
9.Metanephrine-d31.94183.00151.15
10.Metanephrine1.96179.94165.01
11.3-Methoxytyramine-d42.42155.07122.93
12.3-Methoxytyramine2.45151.0091.02
ColumnRaptor Biphenyl (cat.# 9309A62)
Dimensions:150 mm x 2.1 mm ID
Particle Size:2.7 µm
Pore Size:90 Å
Temp.:30 °C
Sample
Conc.: Endogenous level
Inj. Vol.:2 µL
Mobile Phase
A:Water, 0.2% formic acid
B:Methanol, 0.2% formic acid
Time (min)Flow (mL/min)%A%B
0.000.3955
2.500.37525
3.000.3595
3.010.3955
5.000.3955
DetectorMS/MS
Ion Mode:ESI+
Mode:MRM
InstrumentUHPLC
NotesSolid Phase Extraction Procedure:
A 200 µL aliquot of urine sample (Bio-Rad Lyphochek quantitative urine control, normal level) was mixed with 10 µL (1 µg/mL in methanol) internal standard solution and 600 μL of 250 mM ammonium acetate solution. The mixture was loaded onto the EVOLUTE EXPRESS WCX 96-well plate (30 mg), washed with 1 mL water and 1 mL methanol:acetonitrile (60:40), eluted with 200 µL of 5% formic acid in water:methanol (95:5), and injected (2 μL) for analysis.