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Forensic
Applications
Amphetamines, Opiates and Phencyclidine in Oral Fluids
Using 60mg 6mL HyperSep Retain-CX Extraction Column
(Part Number: 60107-308)
Sample Preparation
Add 100 to 500µL of oral fluid sample to a clean tube
Add internal standard(s) and let sit for 10 minutes at
room temperature
Add 800µL of 100mM phosphate buffer (pH= 6.0)
Mix/vortex for 10 seconds
Sample pH should be 6.0±0.5
Adjust pH accordingly with 100mM monobasic or
dibasic sodium phosphate
Condition HyperSep Retain-CX Extraction Column
1 x 200µL CH
3
OH
1 x 200µL DI H
2
O
1 x 200µL 100mM phosphate buffer (pH=6.0)
Apply Sample
Do not exceed 1mL/minute
Wash Column
1 x 500µL DI H
2
O
1 x 500µL 100mM acetic acid
1 x 500µL CH
3
OH
Dry column (5 minutes at >10"Hg)
Elute
1 x 800µL CH
2
Cl
2
/IPA/NH
4
OH (70:26:4)
Do not exceed 1mL/minute
NOTE:
Prepare elution solvent fresh daily; add IPA/NH
4
OH mix,
then add CH
2
Cl
2
(pH 11-12)
For amphetamines and PCP:
Add 100µL of 5% trifluoroacetic acid in methanol after 5 min
Dry for 5 min drying
Evaporate to full dryness at <40°C under a stream of N
2
Derivatize
For Amphetamines*:
Add 50µL PFPA (PFAA)
Mix/vortex
Overlay with N
2
and cap
React 20 minutes at 70°C
Evaporate to dryness at <40°C
Reconstitute with 50µL ethyl acetate
For Opiates*:
Add 200µL of a 1:1 solution of propionic anhydride/pyridine
Make fresh daily
Mix/vortex
React 60 minutes at 40°C
Evaporate to dryness at <40°C
Reconstitute with 50µL ethyl acetate
Analysis
Inject 2µL onto gas chromatograph
* Alternate derivatizations may be used
Phencyclidine does not derivatize