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Vanillylmandelic Acid, Homovanillic Acid, and 5-Hydroxyindoleacetic Acid in Human Urine (Quantitative Control Level 1) on Raptor Biphenyl

LC_CF0661
PeakstR (min)Conc.
(µg/mL)
Precursor IonProduct Ion
1.4-Hydroxy-3-methoxymandelic acid-D3 (VMA-D3)0.900.83200.1139.9
2.Vanillylmandelic acid (VMA)0.922.12–3.90*197.0137.9
3.5-Hydroxyindole-4,6,7-D3-3-acetic-D2 acid (5-HIAA-D5)1.800.83195.1148.0
4.5-Hydroxyindole-3-acetic acid (5-HIAA)1.822.69–4.49*190.0145.9
5.4-Hydroxy-3-methoxyphenyl-D3-acetic-D2 acid (HVA-D5)2.100.83186.1127.0
6.Homovanillic acid (HVA)2.121.33–2.46*181.0121.9
*These concentration ranges were supplied by the vendor (see notes).
ColumnRaptor Biphenyl (cat.# 9309512)
Dimensions:100 mm x 2.1 mm ID
Particle Size:5 µm
Pore Size:90 Å
Guard Column:Raptor Biphenyl guard column cartridge 5 mm, 2.1 mm ID, 5 µm (cat.# 930950252)
Temp.:30 °C
Sample
Diluent:Water
Inj. Vol.:5 µL
Mobile Phase
A:0.1% Formic acid, 5 mM ammonium formate in water
B:Methanol
Time (min)Flow (mL/min)%A%B
0.000.58515
3.000.52080
3.010.58515
5.000.58515
DetectorMS/MS
Ion Mode:ESI-
Mode:MRM
InstrumentUHPLC
NotesThe human urine sample is the Bio-Rad Lyphochek quantitative urine control, level 1. A 40 μL aliquot of urine was mixed with 360 μL of water and 10 μL of internal standard solution (33.3 μg/mL in methanol) in a Thomson 0.45 μm PVDF filter vial and injected for analysis after filtration.