82 / 144
82
Forensic
Applications
Cocaine and Benzoylecgonine and Cocaethylene in Serum, Plasma
Whole Blood, Urine and Tissue
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 whole blood, serum/plasma, urine, or 1g
tissue homogenate (1:4)
Add 2mL of 100mM phosphate buffer (pH 6)
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 HCl
1 x 3mL CH
3
OH Dry column (5 minutes at >10"Hg)
Elute
1 x 3mL ethyl acetate;
ACN:ammonia(78:20:2 v/v)
Collect eluate at 1 to 2mL/minute
Evaporate eluates to dryness under a gentle stream
of nitrogen
Analysis
Reconstitute sample in 50µL of CH
3
OH
Inject 5µL on to LC/MS
Compound
MRM Transition
Cocaine
304.2/182.3
Cocaine-D3*
307.2/185.2
Benzoylecgonine
290.1/168.0
Benzoylecgonine-D8*
298.2/171.3
Cocaethylene
318.2/196.2
Cocaethylene-D8*
326.2/204.2
* Internal standards: Cocaine-D3, Benzoylecgonine-D8, Cocaethylene-D8
Recommended HPLC Column
Part Number
Hypersil GOLD 3µm, 150 x 2.1mm
25003-152130