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Forensic
Applications
Blood GHB Extraction
Using 200mg 3mL HyperSep Retain-AX Extraction Column
(Part Number: 60107-404)
Sample Preparation
To 1mL blood sample add internal standard* and 0.5mL
of 100mM phosphate buffer (pH 6.0)
Mix/vortex
Rock for 10 minutes
Centrifuge for 10 minutes at 2700rpm
Condition HyperSep Retain-AX 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
Place centrifuge tubes into vacuum manifold for collection
The sample loading is collected
Decant sample onto column
Aspirate at about 1"Hg
After the sample is off the columns apply full vacuum for
about 15 seconds to remove any residual blood
Elute
Remove centrifuge tubes, set aside
Place clean centrifuge tubes into vacuum manifold for
collection
1 x 2mL of CH
3
OH/NH
4
OH (99:1)
Aspirate at about 1"Hg
Concentrate
Remove test tubes from vacuum manifold
Mix/vortex the sample prior to concentrating
Evaporate to dryness at 60°C using a stream of nitrogen
Sample Clean-up
Add 200µL of dimethylformamide
Add 1mL of hexane saturated with dimethylformamide
Rock for 5 minutes
Centrifuge at 5 minutes at 2700rpm
Transfer lower dimethylformamide layer to a clean test tube
Evaporate to dryness at 50°C using a stream of air or
nitrogen
Derivatize
Add 25µL ethyl acetate and 25µL BSTFA
(with 1% TMCS**)
Mix/vortex
Heat at 70°C for 30 minutes
Analysis
Inject a 1 to 2µL of the sample onto GC/MS
Compound
Primary Ion *
Secondary
Tertiary
GHB-D6-di-TMS*
239
240
241
GHB-di-TMS
233
234
235
* Suggested internal standard for GC/MS: D6-GHB
** Part number TS-38831
Recommended GC Column
Part Number
TraceGOLD TG-5SilMS 30m x 0.25m x 0.25µm
26096-1420