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Forensic
Applications
Cocaine and its Metabolites from Meconium
for GC or GC/MS Confirmations
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Prepare Sample
Vortex 0.5 to 1g of meconium with 2mL of CH
3
OH
Centrifuge and transfer the supernatant to a clean tube
To each tube add 3mL of 100mM phosphate buffer
(pH 6.0), internal standard and vortex
Matrix must be more aqueous than organic for good
extraction to occur
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
Allow to dry
Wash Column
3mL of DI H
2
O
2mL of 100mM HCl
3mL of CH
3
OH
Dry column (5 minutes at >10"Hg)
Elute Cocaine and Benzoylecgonine
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)
Evaporate
Evaporate the elution solvent to dryness without heating
Derivatize
Add 50µL ethyl acetate and 50µL BSTFA
(with 1% TMCS***)
Overlay with N
2
and cap
Mix/vortex
React for 20 minutes at 70°C
Remove from heat source to cool
NOTE:
Do not evaporate BSTFA solution
Quantitate
Inject 1 to 2µL onto gas chromatograph
For mass spectrometry, monitor the following ions (GC/MS):
Compound
Primary Ion** Secondary
Tertiary
D
3
-Cocaine*
185
201
306
Cocaine
182
198
303
D
3
-Benzoylecgonine-TMS*
243
259
364
Benzoylecgonine-TMS
240
256
361
* Suggested internal standards for GC/MS: D
3
-Cocaine and D
3
-Benzoylecgonine
** 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