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Forensic
Applications
Basic Drugs in Urine, Blood, Plasma/Serum and Tissue
for HPLC Analysis
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Prepare Sample
To 1mL of 100mM phosphate buffer (pH 6.0) add
internal standards
Mix/vortex
Add 1 to 5mL of urine, 1mL of blood, plasma or serum,
or 1g (1:4) tissue homogenate
Mix/vortex
Add 2mL of 100mM phosphate buffer (pH 6.0)
Sample pH should be 6.0±0.5; adjust sample pH accordingly
with 100mM monobasic or dibasic sodium phosphate
Centrifuge as appropriate
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
Wash Column
3mL of DI H
2
O
1mL of 100mM acetic acid
3mL of methanol
Dry column (5 minutes at >10"Hg)
Elute Bases
2mL of CH
4
OH/NH
4
OH (98:2)
Collect eluate at 1 to 2mL/minute
NOTE:
Prepare elution solvent daily
Extract
To the eluate add 2.0mL of DI H
2
O and 500µL of
methylene chloride
Mix/vortex
Centrifuge at 2,000rpm for 10 minutes
Transfer organic lower layer to a clean test tube
Evaporate
Evaporate to dryness at <40°C
Quantitate
Reconstitute in mobile phase and inject onto the
HPLC system