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Phencyclidine in Urine, Blood, Plasma/Serum and Tissue
for GC or GC/MS Confirmations
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Prepare Sample
To 1mL of 100mM phosphate buffer (pH 6.0) add
internal standard(s)*
Add 1mL of urine, blood, plasma/serum or 1g (1:4) of
tissue homogenate
Mix/vortex
Sample pH should be 6.0±0.5
Adjust pH accordingly with 100mM monobasic or dibasic
sodium phosphate
Centrifuge as appropriate
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
3mL of 100mM acetic acid
3mL of CH
3
OH
Dry column (5 minutes at >10"Hg)
Elute Phencyclidine
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 <40°C
Remove immediately upon completion
Reconstitute with 100µL ethyl acetate
Quantitate
Inject 1 to 2µL onto gas chromatograph
For mass spectrometry, monitor the following ions:
Compound
Primary Ion** Secondary
Tertiary
D
5
-phencyclidine*
205
96
247
Phencyclidine
200
91
242
* Suggested internal standard
** Quantitation ion
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
Forensic
Applications
Phencyclidine in Urine, Blood and Plasma/Serum
for LC/MS Confirmations
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Prepare Sample
To 1mL of 100mM phosphate buffer (pH 6.0) add
internal standard(s)*
Add 1mL of urine, blood, plasma/serum
Add 2mL of 100mM phosphate buffer (pH 6.0)
Mix/vortex
Sample pH should be 6.0±0.5
Adjust pH accordingly with 100mM monobasic or dibasic
sodium phosphate
Mix/vortex and centrifuge as appropriate
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
3mL of 100mM acetic acid
3mL of CH
3
OH
Dry column (5 minutes at >10"Hg)
Elute Phencyclidine
3mL of ethyl acetate/acetonitrile/ammonia (78:20:2, v/v/v)
Collect eluate at 1 to 2mL/min
NOTE:
Prepare elution solvent fresh daily
Evaporate
Evaporate to dryness under a stream of nitrogen at <40°C
Reconstitute
Reconstitute sample in 100µL of CH
3
OH
Analysis
Inject 5µL of sample onto LC/MS/MS system
Monitor the following ions:
Compound
MRM Transition
Phencyclidine
244.3/86.1
Phencyclidine-D5*
249.3/86.1
* Suggested internal standard
Mobile phase: acetonitrile/0.1% formic acid (33:67, v/v)
Flow rate: 0.35mL/min
Column temperature: ambient
Recommended HPLC Column
Part Number
Hypersil GOLD 3µm, 150 x 2.1mm
25003-152130