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Forensic
Applications
Amphetamines in Urine, Oxidation with Periodate
for GC or GC/MS Confirmations
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Prepare Sample
To 2mL of urine add internal standard(s)*, 1mL of
100mM phosphate buffer (pH 6.0) and 1mL of
0.35 M sodium periodate
Mix/vortex
Incubate at room temperature for 20 minutes
Sample pH should be 6.0±0.5
Adjust pH accordingly with 100mM monobasic or dibasic
sodium phosphate
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) then aspirate
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
1mL of 100mM acetic acid
3mL of CH
3
OH
Dry column (5 minutes at >10"Hg)
Elute Amphetamines
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)
Concentrate Eluate
Add 30µL silylation grade DMF*** to eluate
Evaporate to 30µL at <40°C
Fluoroacylate with PFPA (PFAA)
Add 50µL PFPA (PFAA)****
Overlay with N
2
and cap
Improve derivatization by addition of 50µL PFPOH
React for 20 minutes at 70°C
Evaporate to dryness at <40°C
Quantitate
Inject 1 to 2µL onto gas chromatograph
For mass spectrometry monitor the following ions:
Analyte (TMS)
Primary Ion** Secondary Tertiary
D
5
-amphetamine*
194
92
123
Amphetamine
190
91
118
D
5
-methamphetamine*
208
92
163
Methamphetamine
204
91
160
* Suggested internal standards
** Quantitation ion
*** Part number TS-20672 (50mL vial)
**** Part number TS-65193 (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