Method for quantification of opioids and their metabolites in autopsy blood by liquid chromatography-tandem mass spectrometry.
A method using liquid chromatography-electrospray ionization-tandem mass spectrometry was developed and validated for the determination of morphine, codeine, hydromorphone, dihydrocodeine, oxycodone, buprenorphine, and naloxone with their metabolites morphine-3-glucuronide, morphine-6-glucuronide, normorphine, 6-acetylmorphine, 6-acetylcodeine, codeine-6-glucuronide, norcodeine, hydromorphine-3-glucuronide, dihydrocodeine-6-glucuronide, dihydromorphine, dihydromorphine-3-glucuronide, dihydromorphine-6-glucuronide, oxymorphone, norbuprenorphine, buprenorphine-3-glucuronide, norbuprenorphine-3-glucuronide, and naloxone-3-glucuronide in human whole blood. Polar metabolites (glucuronides) and other analytes were extracted by SPE using Bond Elut C18. Chromatographic separation was performed on a Phenomenex Synergi reversed-phase column with gradient elution based on a mobile phase consisting of 10mM ammonium formate adjusted to pH 3 and acetonitrile. Intraday and interday precision for all analytes were between 0.6% and 13.8%, and recoveries were between 80.3% and 101.4%. Calibration curves were linear for all analytes over the concentration range 5-400 ng/mL, and correlation coefficients (R(2)) were better than 0.999. Limits of detection and quantitation were 0.16-1.2 ng/mL and 0.5-4.09 ng/mL, respectively. The method described consolidates previous work on opioids and their metabolites published in the literature and is the first to include the detection of naloxone-3-glucuronide. The method has been applied in routine postmortem cases after opiate overdose with the threefold purpose of providing interpretive information on the cause and type of death (rapid, sub-acute, or delayed death) and to distinguish heroin, morphine, and codeine users.
Al-Asmari
Ahmed I
AI
Forensic Medicine and Science Department, University of Glasgow, University Place, G12 8QQ, Scotland. Ahmadalasmari@yahoo.com
Anderson
Robert A
RA
D000701:Analgesics, Opioid / Q000097:blood* / Q000378:metabolism / Q000506:poisoning; D001344:Autopsy; D002138:Calibration; D002423:Cause of Death; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005554:Forensic Medicine; D020719:Glucuronides / Q000097:blood; D006801:Humans; D053610:Opiate Alkaloids / Q000097:blood* / Q000378:metabolism / Q000506:poisoning; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D015813:Substance Abuse Detection / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
D000701:Analgesics, Opioid; D020719:Glucuronides; D053610:Opiate Alkaloids