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Forensic
Applications
6-Acetylmorphine in Urine
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Sample Preparation
1 x 2mL of 100mM phosphate buffer (pH= 6.0)
Add internal standard*
Mix/vortex
Add 4mL of sample
Centrifuge for 10 minutes at 2,000rpm
Discard pellet
Sample pH should be 6.0±0.5
Adjust pH to 6 with 100mM monobasic or dibasic
sodium phosphate
Condition HyperSep Verify-CX Extraction Column
1 x 3mL CH
3
OH
1 x 3mL DI H
2
O
1 x 1mL 100mM phosphate buffer (pH=6.0)
NOTE:
Aspirate at <3"Hg to prevent sorbent drying
Apply Sample
Load at 1mL/minute
Wash Column
1 x 3mL DI H
2
O
1 x 2mL 100mM acetate buffer (pH 4.5)
1 x 3mL CH
3
OH
Dry column (10 minutes at >10"Hg)
Elute
1 x 3mL CH
2
Cl
2
/IPA/NH
4
OH (78:20:2)
Collect eluate at 1 to 2mL/minute
NOTE:
Prepare elution solvent fresh daily; add IPA/NH
4
OH mix,
then add CH
2
Cl
2
(pH 11-12)
Evaporate to dryness at <40°C
Derivatize
Add 50µL ethyl acetate
Mix/vortex
Add 50µL BSTFA (with 1% TMCS)**
Overlay with N
2
and cap
Mix/vortex
React 45 minutes at 70°C
Remove from heat source to cool
NOTE:
Do not evaporate BSTFA solution
Analysis
Inject 1 to 2µL onto GC/MS
For mass spectrometry analysis monitor the following ions:
Compound
Primary Ion*** Secondary
Tertiary
D6-6-AM-TMS*
405
406
343
6-AM-TMS
399
400
340
* Suggested internal standard for GC/MS:
** Part number TS-38831
*** Quantitation ion
Recommended GC Column
Part Number
TraceGOLD TG-5SilMS 30m x 0.25m x 0.25µm
26096-1420