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Forensic
Applications
Sympathomimetic Amines in Urine, Blood, Plasma/Serum and
Tissue for GC or GC/MS Confirmations
Alternative Drying Procedure – Fluoroacylate PFPA (PFAA) Derivative
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Prepare Sample
To 1mL of 100mM phosphate buffer (pH 6.0) internal
standard (s)*
Add 1mL of urine, blood, plasma/serum or 1g of (1:4)
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) 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 SMA
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
Alternate Drying Procedure – Fluoroacylate PFPA (PFAA)
Derivative
Add 50µL PFPA (PFAA)****
Overlay with N
2
and cap
Improved derivatization by addition of 50µL PFPOH
†
React for 20 minutes at 70°C
Evaporate to dryness at <40°C
Reconstitute with 100µL ethyl acetate
Quantitate
Inject 1 to 2µL onto gas chromatograph
For mass spectrometry, monitor the following ions:
Analyte (PFPA)
Primary Ion** Secondary Tertiary
D
5
-amphetamine*
194
92
123
Amphetamine
190
91
118
D
5
-methamphetamine*
208
92
163
Methamphetamine
204
91
160
Pseudoephedrine
204
160
119
Ephedrine
204
160
119
Phenylephrine
190
119
267
Methylenedioxyamphetamine 135
162
325
Methylenedioxymethamphetamine 204
162
339
* Suggested internal standards for GC/MS: D
5
-amphetamine and D
5
-methamphetamine
** Quantitation ion
*** Part number TS-20672 (50mL vial)
**** Part number TS-65193 (10 x 1mL ampules)
† Part number TS-65195 (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