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Forensic
Applications
Beta Blockers in Urine and Blood for GC/MS Confirmations
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Prepare Sample
To 1mL of urine or blood add 1mL of 100mM acetate
buffer (pH 4.5)
Add 2mL of acetate buffer (pH 4.5)
Mix/vortex
Centrifuge as appropriate
Condition Verify-CX Extraction Column
3mL of CH
3
OH then aspirate
3mL of DI H
2
O then aspirate
3mL of 100mM acetate buffer (pH 4.5) then aspirate
NOTE:
Aspirate at <3"Hg to prevent sorbent drying
Apply Sample
Load sample at 1 to 2mL/minute
Wash Column
2 x 1mL of acetone/methanol (1:1) then aspirate
Dry column (5 minutes at >10"Hg)
Elute Beta Blockers
1mL of 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)
Dry Eluate
Evaporate to dryness at <40°C
Derivatize
Derivatization solution:
Methaneboronic acid at 5mg/mL
prepared in dry ethyl acetate (use molecular sieve)
Store this solution at -20°C (freezer conditions) until use
Reaction mixture:
Add 100µL of the methaneboronic
acid solution (see above)
Mix/vortex
React for 15 minutes at 70°C
Remove from heat source to cool
NOTE:
Do not evaporate this solution
Analysis
Inject 1 to 2µL of sample
Beta Agonists in Urine
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Sample Preparation
To 1mL of 100mM acetate buffer (pH 4.5)
Add 1mL of urine
Add 2mL of 100mM acetate buffer (pH 4.5)
Mix/vortex
Centrifuge as appropriate
Condition HyperSep Verify-CX Extraction Column
1 x 3mL CH
3
OH
1 x 3mL DI H
2
O
1 x 3mL 100mM acetate buffer (pH 4.7)
NOTE:
Aspirate at <3"Hg to prevent sorbent drying
Apply Sample
Load at 1 to 2mL/minute
Wash Column
2 x 1mL acetone/methanol (1:1) aspirate
Dry column (5 minutes at >10"Hg)
Elute
1 x 1mL dichloromethane/isopropanol and ammonium
hydroxide (78:20:2)
Collect the eluate at 1 to 2mL/minute (or gravity)
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
Derivatize
Derivatization solution: Methaneboronic acid at 5mg/mL
prepared in dry ethyl acetate (use molecular sieve)
Store this solution at –20°C (freezer conditions) until use
Reaction Mixture
Add 100µL of the methaneboronic acid solution (see above)
Mix/vortex
React 15 minutes at 70°C
Remove from heat source to cool
NOTE:
Do not evaporate this solution
Analysis
Inject 1 to 2µL sample (derivatized solution)