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Forensic
Applications
Meperidine and Normeperidine in Blood, Plasma, Serum and Urine
Using 200mg 3mL HyperSep Aminopropyl Extraction Column
(Part Number: 60108-425)
Sample Preparation
1mL of 100mM phosphate buffer (pH 6)
Add internal standard*
Add 1mL of whole blood, serum/plasma, urine
Add 2mL of 100mM phosphate buffer (pH 6)
Mix/vortex
Centrifuge
Condition HyperSep Aminopropyl 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 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 onto LC/MS
Mobile phase:
Time %
ACN %
0.1% Formic Acid
0
90
10
5
30
70
6
90
10
10
90
10
Flow rate: 0.35mL/minute
Column temperature: ambient
Detector: Thermo Scientific TSQ Triple Quadrupole
Compound
MRM Transition
Meperidine
248.2/220.0
Meperidine-D4*
252.2/224.1
Normeperdidine
234.1/160.0
Normeperidine-D4*
238.1/164.0
* Suggested internal standard: Meperidine-D4 and Normeperidine-D4
Recovery: >90% (n=10)
LOD: 10ng/mL
Recommended HPLC Column
Part Number
Hypersil GOLD 3µm, 150 x 2.1mm
25003-152130