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Forensic
Applications
Delta 9-THC, Delta 9-Hydroxy THC, Carboxy-Delta-9-THC in Whole Blood
Using 200mg 6mL HyperSep Verify-AX Extraction Column
(Part Number: 60108-730)
Sample Preparation
1 to 2mL whole blood
Add internal standards* prepared in alcohol
Add drop-wise 2mL ice cold ACN
Mix thoroughly and centrifuge
Decant ACN into a clean tube
Evaporate ACN under a stream or air or nitrogen
to 200µL
Add 2mL distilled H
2
O (pH~6.0-7.0)
NOTE:
The ACN should be cold and added slowly to prevent
precipitation
Apply Sample
Load sample directly to column without any
preconditioning
Wash Column
Wash with 1mL (84/15/1) water/ACN/NH
4
OH
Dry column thoroughly under vacuum
NOTE:
It is important to dry the column properly to achieve the
highest recovery of compounds
Elute
1 x 3mL hexane/ethyl acetate/glacial acetic acid (49:49:2)
Collect at 1 to 2mL/minute
Evaporate fraction(s) to dryness under stream of dry air
or nitrogen at <40°C
Derivatize
Add 50µL ethyl acetate
Mix/vortex
Add 50µL BSTFA (with 1% TMCS)***
React 20 minutes at 70°C
Remove from heat source to cool
NOTE:
Do not evaporate BSTFA
Analysis
Inject 2µL onto GC/MS
For mass spectrometry monitor the following ions:
Derivatization procedure:
Derivatizing
THC {T-005}**
THC-OH {H-041}**
THC-COOH {T-006}**
Agent
(D3 THC) {T-003}**
(D3 THC-OH) {H-027}**
(D9 THC-COOH) {T007}**
BSTFA
371, 343, 386
371, 459, 474
371, 473, 488
(374, 346, 389)
(374, 462, 477)
(380, 479, 497)
* Suggested internal standard for GC/MS: D9-Carboxy-delta 9-THC, D3-Hyroxy- delta 9-THC, D3-delta 9-THC
** Ions common to deuterated delta-9 THC and non-deuterated compounds
*** Part number TS-38831
Recommended GC Column
Part Number
TraceGOLD TG-35MS 30m x 0.25m x 0.25µm
26094-1420