Forensic Toxicology

, Volume 37, Issue 1, pp 121–131 | Cite as

Simultaneous determination of selegiline, desmethylselegiline, R/S-methamphetamine, and R/S-amphetamine in oral fluid by LC/MS/MS

  • Lizhu Chen
  • Yingjia Yu
  • Yang Wang
  • Ping Xiang
  • Gengli Duan
Original Article



It is crucial for forensic analysis to differentiate clinical use from illegal abuse. Selegiline (SG) is mainly metabolized to desmethylselegiline (DM-SG), R-(−)-methamphetamine (R-MA) and R-(−)amphetamine (R-AM); while abused methamphetamine and amphetamine mainly contain the S-(+)-form. The aim of this study was to simultaneously determine SG, DM-SG, R/S-MA, and R/S-AM in human oral fluid (OF) and to differentiate clinical use from illegal use. We also aim to apply the present method to the OF samples from authentic cases in forensic toxicology.


Liquid–liquid extraction and liquid chromatography-tandem mass spectrometry operating in positive ion multiple-reaction monitoring mode were utilized. The chromatographic system consisted of a ChirobioticTM V2 column (2.1 mm × 250 mm, 5 μm) and a mobile phase of methanol containing 0.1% (v/v) glacial acetic acid and 0.02% (v/v) ammonium hydroxide.

Results and conclusions

The calibration curves were linear from 1 to 100 ng/mL, and r > 0.995 for all analytes with imprecisions ≤ 15% and accuracy between 85 and 115%. Extraction recoveries ranged from 46.3 to 104.7% with coefficient of variation (CV) ≤ 10.3% and matrix effects (MEs) ranged from 47.4 to 114.8% with CV ≤ 12.9%. The lower limit of quantification was 0.2 ng/mL for SG and DM-SG and 1.0 ng/mL for S-MA, R-MA, S-AM and R-AM. The present method is simple, rapid (accomplished in 12 min), sensitive, and validated by authentic samples.


Oral fluid Chiral analysis R/S-methamphetamine R/S-amphetamine Selegiline Desmethylselegiline 



The authors are grateful to the Ministry of Science and Technology (2016YFC0800704), the National Natural Science Foundation of China (81772022, 81871531), the Science and Technology Commission of Shanghai Municipality (17DZ2273200/16DZ2290900), and Ministry of Finance, PR China (GY2017G-1) for their financial support of this study.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in this study involving human participants were in accordance with the ethical standards of the international and/or national committee and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.


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Copyright information

© Japanese Association of Forensic Toxicology and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Pharmaceutical Analysis, School of PharmacyFudan UniversityShanghaiChina
  2. 2.Department of Forensic Toxicology, Ministry of Justice, Shanghai Key Laboratory of Forensic MedicineAcademy of Forensic SciencesShanghaiChina

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