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Forensic
Applications
Cocaine and Benzoylecgonine in Serum, Plasma and Whole Blood
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Sample Preparation
4mL of DI H
2
O
Add internal standards
1mL of sample (Serum, Plasma or Whole Blood)
Add internal standard(s)
Mix/vortex and let stand 5 minutes
Centrifuge for 10 minutes at 2,000rpm and discard pellet
Add 2mL of 100mM phosphate buffer (pH =6.0)
Mix/vortex
Sample pH should be 6.0
Adjust pH 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 1 to 2mL/minute
Wash Column
1 x 3mL DI H
2
O
1 x 2mL 100mM HCl
1 x 3mL CH
3
OH
Dry column (5 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
Analysis
Reconstitute in mobile phase for injection into HPLC
Recommended HPLC Column
Part Number
Hypersil GOLD 3µm, 150 x 4.6mm
25003-154630