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Forensic
Applications
Fentanyl and Analogues 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 standards*
Add 1 to 5mL of urine, blood, plasma/serum, or 1g (1:4)
of tissue homogenate
Mix/vortex
Add 2mL of 100mM phosphate buffer (pH 6.0)
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 Fentanyls
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
Evaporate to dryness at <40°C
Reconstitute with 50µL ethyl acetate
Quantitate
Inject 1 to 2µL onto gas chromatograph
For mass spectrometry, monitor the following ions:
Compound
Primary Ion** Secondary
Tertiary
Fentanyl
245
146
189
D
5
-Fentanyl*
250
151
194
α
-Methylfentanyl
259
203
146
Para-Fluorofentanyl
263
164
207
3-Methylfentanyl
259
160
203
Thienfentanyl
245
146
189
Sufentanil
289
140
Carfentanil
303
187
Lofentanil
317
201
289
Alfentanil
289
268
194
* 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