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Psilocin and Psilocybin in Urine on Raptor Biphenyl by Restek's 'Big Pain' Method (CFAR2309-UNV)

LC_CF0772
PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct Ion 1Product Ion 2
1.Psilocybin2.09500285.1205.1240.0
2.Psilocin-D102.74200215.166.1-
3.Psilocin2.78500205.1160.1115.0
ColumnRaptor Biphenyl (cat.# 9309A12)
Dimensions:100 mm x 2.1 mm ID
Particle Size:2.7 µm
Pore Size:90 Å
Guard Column:Raptor Biphenyl EXP guard column cartridge 5.0 mm, 2.1 mm ID, 2.7 µm (cat.# 9309A0252)
Temp.:30 °C
Sample
Diluent:Water, 0.1% formic acid + 2 mM ammonium formate
Conc.:500 ng/mL
Inj. Vol.:5 µL
Mobile Phase
A:Water, 0.1% formic acid + 2 mM ammonium formate
B:Methanol, 0.1% formic acid + 2 mM ammonium formate
Time (min)Flow (mL/min)%A%B
0.000.6955
9.000.60100
10.000.60100
10.010.6955
12.000.6955
DetectorMS/MS
Ion Source:Electrospray
Ion Mode:ESI+
Mode:MRM
InstrumentHPLC
NotesA 500 ng/mL standard mix of psilocin and psilocybin was prepared in pooled urine. A 50 µL aliquot was taken from the standard and mixed with 10 µL of internal standard (psilocin-D10, 20 µg/mL) and 100 µL of methanol. The mixture was vortexed at 3000 rpm for 10 seconds and centrifuged at 4300 rpm for 10 minutes at 10 °C. After centrifugation, 100 µL of the supernatant was diluted with 900 µL (20-fold dilution) of water containing 0.1% formic acid and 2 mM ammonium formate (mobile phase A) and injected for LC-MS/MS analysis.