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Forensic
Applications
Duloxetine in Blood and Urine
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Sample Preparation
1mL of 100mM phosphate buffer (pH= 6)
Add internal standard*
Add 1mL of blood or urine
Add 2mL of 100 phosphate buffer (pH= 6)
Mix/vortex
Sample pH should be 6.0±0.5
Adjust pH with 100mM monobasic or dibasic
sodium phosphate
Mix/vortex
Centrifuge
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)
NOTE:
Aspirate at <3"Hg to prevent sorbent drying out
Apply Sample
Load sample at 1 to 2mL/minute
Wash Column
1 x 3mL DI H
2
O
1 x 3mL 100mM acetic acid
1 x 3mL CH
3
OH
Dry column (5 minutes at >10"Hg)
Elute
1 x 3mL dichloromethane/isopropanol/ammonia (78:20:2 v/v)
Collect eluate at 1 to 2mL/minute
Evaporate eluate under a gentle stream of nitrogen <40ºC
Analysis
Reconstitute sample in 200µL of 0.1% formic acid
Inject 5µL onto LC/MS
Compound
MRM Transition
Ethyl Morphine*
314.2/152.2
Duloxetine
298.1/44.1
* Internal standard: Ethyl Morphine