Direct Injection LC-MS-MS Analysis of Opiates, Methamphetamine, Buprenorphine, Methadone and Their Metabolites in Oral Fluid from Substitution Therapy Patients.
A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS-MS) method was developed, validated and applied to simultaneous analysis of oral fluid samples for the following 10 analytes: methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), buprenorphine, norbuprenorphine, morphine, codeine, 6-acetylmorphine, 6-acetylcodeine, amphetamine, and methamphetamine. The oral fluid sample was briefly centrifuged and the supernatant was directly injected into the LC-MS-MS system operated under reverse-phase chromatography and electrospray ionization (ESI). Deuterated analogs of the analytes were adopted as the internal standards and found to be effective (except for buprenorphine) to compensate for potential matrix effects. Each analytical run took <10 min. Linearity range (r(2) > 0.99) established for buprenorphine and the other nine analytes were 5-100 and 1-100 ng/mL. Intra- and interday precision (% CV) ranges for the 10 analytes were 0.87-12.2% and 1.27-12.8%, while the corresponding accuracy (%) ranges were 91.8-113% and 91.9-111%. Limits of detection and quantitation established for these 10 analytes were in the ranges of 0.1-1.0 and 0.25-1.0 ng/mL (5 ng/mL for buprenorphine). The method was successfully applied to the analysis of 62 oral fluid specimens collected from patients participating in methadone and buprenorphine substitution therapy programs. Analytical results of methadone and buprenorphine were compared with data derived from GC-MS analysis and found to be compatible. Overall, the direct injection LC-MS-MS method performed well, permitting rapid analysis of oral fluid samples for simultaneous quantification of methadone, buprenorphine, opiate and amphetamine drug categories without extensive sample preparation steps.
Department of Forensic Toxicology, Institute of Forensic Medicine, Ministry of Justice, Taipei, Taiwan.
Hsiu-Chuan
HC
Liu
Department of Forensic Toxicology, Institute of Forensic Medicine, Ministry of Justice, Taipei, Taiwan.
Hsi-Tzu
HT
Lee
Department of Medical Laboratory Science and Biotechnology, Fooyin University, Kaohsiung, Taiwan.
Ya-Ching
YC
Hsu
Department of Medical Laboratory Science and Biotechnology, Fooyin University, Kaohsiung, Taiwan.
Mei-Han
MH
Huang
Department of Justice Sciences, University of Alabama at Birmingham, Birmingham, AL, USA.
Ray H
RH
Liu
Department of Psychiatry, E-Da Hospital, Kaohsiung, Taiwan.
Tai-Jui
TJ
Chen
Department of Forensic Toxicology, Institute of Forensic Medicine, Ministry of Justice, Taipei, Taiwan dllin@mail.moj.gov.tw.
Dong-Liang
DL
Lin
D000697:Central Nervous System Stimulants / Q000032:analysis*; D002853:Chromatography, Liquid; D056148:Chromatography, Reverse-Phase; D006801:Humans; D009294:Narcotics / Q000032:analysis*; D053610:Opiate Alkaloids / Q000032:analysis*; D058850:Opiate Substitution Treatment; D012463:Saliva / Q000737:chemistry*; D015813:Substance Abuse Detection / Q000379:methods*; D013624:Taiwan; D053719:Tandem Mass Spectrometry
D000697:Central Nervous System Stimulants; D009294:Narcotics; D053610:Opiate Alkaloids