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Psilocin in Urine for GC or GC/MS Confirmations
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Prepare Sample
To 5mL of urine add internal standard* (Psilocin-D10-TMS)
and 2mL of 100mM phosphate buffer (pH 6.0)
Mix/vortex
Add 12,500 to 25,000 units of
β
-glucuronidase, then
mix/vortex
Place the sample into a water bath at 45°C for 90 minutes
Remove from the bath and allow to cool
Centrifuge at 3,000rpm for 10 minutes
Use the clear filtrate (discard the plug) for SPE
Condition Verify-CX Extraction Column
3mL of CH
3
OH then aspirate
3mL of DI H
2
O then aspirate
1mL of 100mM phosphate buffer (pH 6.0)
NOTE:
Aspirate at <3"Hg to prevent sorbent drying
Apply Sample
Load sample at 1 to 2mL/minute
Wash Column
3mL of DI H
2
O
2mL of 20% acetonitrile in water
1mL of 100mM acetic acid
Dry column (3 minutes at >10"Hg)
2mL of hexane
3mL of hexane/ethyl acetate (50:50)
3mL of CH
3
OH
Dry column (3 minutes at >10"Hg)
Elute Psilocin
3mL of CH
2
Cl
2
/IPA/NH
4
OH (78:20:2)
Collect eluate at 1 to 2mL/min
NOTE:
Prepare elution solvent fresh daily; add IPA/NH
4
OH mix,
then add CH
2
Cl
2
(pH 11-12)
Dry Eluate
Evaporate to dryness at <35°C
Derivatize
Add 50µL of ethyl acetate
Mix/vortex
Add 50µL of MSTFA***
React at 70°C for 30 minutes
Remove from heat
NOTE:
Do not evaporate MSTFA solution
Quantitate
Inject 1 to 2µL onto gas chromatograph
For mass spectrometry, monitor the following ions:
Compound
Primary Ion** Secondary
Tertiary
PSILOCIN-TMS
290
348
73 (291)
PSILOCIN-D10-TMS*
300
358
83 (301)
* Suggested internal standard
** Quantitation ion
*** Part number TS-48910 (10 x 1mL ampules)
Recommended GC Columns
Part Number
TRACE TR-DoA5 Column, 15m x 0.25mm x 0.25µm
26AF130P
TRACE TR-DoA35 Column, 15m x 0.20mm x 0.33µm
26AC497P
Source
Greishaber A., Moore, K. Levine, B, and Smith M. (1999, November). The detection of
psilocin in human urine. Presented at the TRI Services Meeting.
Forensic
Applications