Determination of buprenorphine and norbuprenorphine in whole blood by liquid chromatography-mass spectrometry.
A sensitive and highly specific method for the determination of buprenorphine and norbuprenorphine in postmortem and hemolyzed whole blood using combined liquid chromatography and mass spectrometry (LC-MS) with electrospray ionization was developed. After enzymatic hydrolysis and deproteinization with acetonitrile, Extrelut-3 cartridges were used for a preliminary basic sample cleanup. Elution by toluene-ether (50:50, v/v) was followed by an acid wash with 0.05M H3PO4 and a basic re-extraction into ether. A Nucleosil C18 (150 x 1-mm i.d.) reversed-phase column was used for the chromatographic separation together with a mixture of 2mM ammonium formate buffer (pH 3) and acetonitrile (55:45, v/v) as the mobile phase. Recoveries were between 56 and 60%, and detection limits were 0.05 ng/mL for both analytes. The coefficient of variation for repeatability was lower than 4%. Limits of quantitation were 0.1 ng/mL for buprenorphine and norbuprenorphine. Reproducibility was good, and linearity was excellent in the range of 0.1 to 100 ng/mL (r > 0.9999, n = 7).
Hoja
H
H
Department of Pharmacology and Toxicology, University Hospital, Limoges, France.
Marquet
P
P
Verneuil
B
B
Lotfi
H
H
Dupuy
J L
JL
Lachâtre
G
G
D000328:Adult; D002047:Buprenorphine / Q000031:analogs & derivatives* / Q000097:blood* / Q000652:urine; D002853:Chromatography, Liquid; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D009294:Narcotics / Q000097:blood* / Q000652:urine; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D009294:Narcotics; D002047:Buprenorphine; C043585:norbuprenorphine