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Propoxyphene and Norpropoxyphene in Urine, Blood,
Plasma/Serum and Tissue for LC/MS/MS Confirmations
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 standard*
Add 1mL of urine, blood, plasma/serum, or 1g (1:4) of
tissue homogenate
Add 2mL of 100mM phosphate buffer (pH 6.0)
Vortex and 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)
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
3mL of 100mM acetic acid
3mL of CH
3
OH
Dry column (5 minutes at >10"Hg)
Elute Propoxyphene/Norpropoxyphene
3mL of ethyl acetate/acetonitrile/ammonia
(78:20:2, v/v/v) OR
3mL of CH
2
Cl
2
/IPA/NH
4
OH (78:20:2, v/v/v)
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 to half volume under a stream of nitrogen
at <40°C
Add 100µL of 0.1% HCl in CH
3
OH
Mix/vortex
Continue evaporation to dryness at <40°C
Reconstitute
Reconstitute sample in 100µL of CH
3
OH
Analysis
Inject 5µL of sample onto LC/MS/MS system
Monitor the following ions:
Compound
MRM Transition
Propoxyphene
340.0/58.0
Propoxyphene-D11*
351.2/64.0
Norpropoxyphene
326.0/252.0
Norpropoxyphene-D5*
331.0/257.0
Flow rate: 0.35mL/min
Column temperature: ambient
Mobile phase:
Time (min)
% Acetonitrile
% of 0.1% Formic Acid
0
30
70
10
30
70
Recommended HPLC Column
Part Number
Hypersil GOLD Phenyl 3µm, 50 x 2.1mm
25903-052130
Forensic
Applications