HPLC-DAD determination of opioids, cocaine and their metabolites in plasma.
High performance liquid chromatography with diode array detection (HPLC-DAD) was used to develop a method for the simultaneous determination of morphine, codeine, 6-acetylmorphine (6AM), cocaine, benzoylecgonine (BEG), cocaethylene, methadone and its metabolite, 2-ethylidene-1,5-dimethyldiphenylpyrrolidine (EDDP), in plasma. Following solid-phase extraction with Bond Elut Certify cartridges, chromatography was performed on an X-Terra RP8 column (250 mm x 4.6 mm i.d., 5 microm particle size), using acetonitrile-phosphate buffer pH 6.53 as mobile phase and elution in the gradient mode. The detector response was linear at concentrations over the range 0.1-10 microg/mL in plasma, and the correlation coefficients for the eight drugs studied were all higher than 0.99. The average extraction recoveries from plasma ranged from 60% for BEG to 95% for methadone. The precision was acceptable, with coefficients of variation oscillating between 2.55% and 6.45%. The accuracy was found to be within satisfactory limits (+/- 8.1%). Finally, the method was applied to 21 plasma samples from fatal overdoses, obtaining positive results for two or more drugs.
Fernández
P
P
Institute of Legal Medicine, Forensic Toxicology Service, Faculty of Medicine, University of Santiago de Compostela, C/San Francisco s/n, 15782 Santiago de Compostela, Spain.
Morales
L
L
Vázquez
C
C
Bermejo
A M
AM
Tabernero
M J
MJ
D000701:Analgesics, Opioid / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003042:Cocaine / Q000031:analogs & derivatives / Q000097:blood*; D003061:Codeine / Q000097:blood; D018765:Dopamine Uptake Inhibitors / Q000097:blood*; D005260:Female; D005554:Forensic Medicine; D006801:Humans; D008297:Male; D008691:Methadone / Q000097:blood; D009020:Morphine / Q000097:blood; D009022:Morphine Derivatives / Q000097:blood; D011759:Pyrrolidines / Q000097:blood
D000701:Analgesics, Opioid; D018765:Dopamine Uptake Inhibitors; D009022:Morphine Derivatives; D011759:Pyrrolidines; C005618:benzoylecgonine; D009020:Morphine; C066444:cocaethylene; D003042:Cocaine; C026979:6-O-monoacetylmorphine; D008691:Methadone; D003061:Codeine; C010724:2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine