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Tetrahydrocannabinol (THC) in Oral Fluid for GC/MS Confirmation
Using 50mg 1mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-741)
Prepare Sample
Add 100 to 500µL of neat sample to a clean test tube
Add internal standard*
Vortex and let sit for 10 minutes at room temperature
Add 0.5mL of glacial acetic acid and mix/vortex for
10 seconds
Condition Verify-CX Extraction Column
200µL of CH
3
OH
200µL of DI H
2
O
200µL of 100mM HCl
Apply Sample
Load sample at 1mL/minute (do not exceed this flow rate)
Wash Column
500µL of DI H
2
O
500µL of 0.2N HCl
500µL of 100mM HCl/acetonitrile (70:30)
Dry column (1 minute at >10"Hg)
Elute THC
800µL of hexane/ethyl acetate (75:25)
Collect eluate at 1mL/min (do not exceed this rate)
Dry Eluate
Evaporate to full dryness at <40°C under a stream
of nitrogen
Derivatize
Add 25µL of BSTFA (with 1% TMCS)*** and
25µL of ethyl acetate
Overlay with nitrogen and cap
Vortex then react for 30 minutes at 70°C
Remove from heat and allow to cool
NOTE:
Do not evaporate BSTFA solution
Quantitate
Inject 2µL onto gas chromatograph
Monitor the following ions:
Compound
Primary Ion** Secondary
Tertiary
THC-TMS
371
386
303
THC D3-TMS*
374
389
318
Sample is from either a neat sample capillary tube collection, or eluted off
the cotton pad of a swab collection device with oral fluid THC buffer
* Suggested internal standard
** Quantitation ion
*** Part number TS-38831 (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
122
Forensic
Applications
THC in Oral Fluid
Using 60mg 6mL HyperSep Retain-CX Extraction Column
(Part Number: 60107-308)
Sample Preparation
Add 100 to 500µL of neat oral fluid sample to a clean tube
Add internal standard*
Mix/vortex and let sit for 10 minutes at room temperature
Add 500µL of glacial acetic acid
Mix/vortex for 10 seconds
Condition HyperSep Retain-CX Extraction Column
1 x 200µL CH
3
OH
1 x 200µL DI H
2
O
1 x 200µL 100mM HCl
Apply Sample
Do not exceed 1mL/minute
Wash Column
1 x 500µL DI H
2
O
1 x 500µL 0.2 N HCl
1 x 500µL 100mM HCl/ACN (70:30)
Dry column (1 minute at >10"Hg)
Elute
1 x 800µL ethyl acetate/hexane (25:75)
Do not exceed 1mL/minute
Evaporate at <40°C under a stream of N
2
Derivatize
Add 25µL BSTFA (with 1% TMCS)** and 25µL ethyl acetate
Overlay with N
2
and cap
Mix/vortex
React 30 minutes at 70°C
Remove from heat and allow to cool
NOTE:
Do not evaporate BSTFA solution
Analysis
Inject 2µL onto GC/MS
Monitor the following ions:
Compound
Primary Ion
Secondary
Tertiary
THC-TMS
371
386
303
THC-D3-TMS*
374
389
318
* Suggested internal standard: THC-D3-TMS
** Part number TS-38831
Recommended GC Column
Part Number
TraceGOLD TG-35MS 30m x 0.25m x 0.25µm
26094-1420