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Development of a rapid-fire drug screening method by probe electrospray ionization tandem mass spectrometry for human urine (RaDPi-U)

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Abstract

Drug screening tests are mandatory in the search for drugs in forensic biological samples, and immunological methods and mass spectrometry (e.g., gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry) are commonly used for that purpose. However, these methods have some drawbacks, and developing new screening methods is required. In this study, we develop a rapid-fire drug screening method by probe electrospray ionization tandem mass spectrometry (PESI-MS/MS), which is an ambient ionization mass spectrometry method, for human urine, named RaDPi-U. RaDPi-U is carried out in three steps: (1) mixing urine with internal standard (IS) solution and ethanol, followed by vortexing; (2) pipetting the mixture onto a sample plate for PESI; and (3) rapid-fire analysis by PESI-MS/MS. RaDPi-U targets 40 forensically important drugs, which include illegal drugs, hypnotics, and psychoactive substances. The analytical results were obtained within 3 min because of the above-mentioned simple workflow of RaDPi-U. The calibration curves of each analyte were constructed using the IS method, and they were quantitatively valid, resulting in good linearity (0.972–0.999) with a satisfactory lower limit of detection and lower limit of quantitation (0.01–7.1 ng/mL and 0.02–21 ng/mL, respectively). Further, both trueness and precisions were 28% or less, demonstrating the high reliability and repeatability of the method. Finally, we applied RaDPi-U to three postmortem urine specimens and successfully detected different drugs in each urine sample. The practicality of the method is proven, and RaDPi-U will be a strong tool as a rapid-fire drug screening method not only in forensic toxicology but also in clinical toxicology.

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Acknowledgements

We are very grateful to Dr. Y. Hayashi, Ms. M. Shibata, and Ms. K. Furui for their assistance with data acquisition and sample preparation, and Dr. Issey Takahashi for making a great graphical abstract.

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Authors and Affiliations

  1. Forensic Science Laboratory, Aichi Prefectural Police Headquarters, 2-1-1, Sannomaru, Naka-ku, Nagoya, 460-8502, Japan

    Kazuaki Hisatsune

  2. Shimadzu Corporation, 1, Nishinokyo-Kuwabaracho Nakagyo-ku, Kyoto, 604-8511, Japan

    Tasuku Murata & Koretsugu Ogata

  3. Nagoya City Public Health Research Institute, Sakurazaka, Moriyama-ku, Nagoya, 463-8585, Japan

    Masaru Taniguchi

  4. Department of Human Life and Environment, Kinjo Gakuin University, 2-1723 Omori, Moriyama-ku, Nagoya, 463-8521, Japan

    Tomomi Asano

  5. Geological Survey of Japan, National Institute of Advanced Industrial Science and Technology (AIST), AIST Tsukuba Central 7, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8567, Japan

    Akira Iguchi

  6. Research Laboratory On Environmentally-conscious Developments and Technologies [E-code], National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, 305-8567, Japan

    Akira Iguchi

  7. Multimodal Informatics and Wide-Data Analytics Laboratory (MiWA-Lab.), Department of Computational Systems Biology, Faculty of Biology-Oriented Science and Technology, Kindai University, 930 Nishi Mitani, Kinokawa, Wakayama, 649-6493, Japan

    Kei Zaitsu

  8. In Vivo Real-Time Omics Laboratory, Institute for Advanced Research, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8601, Japan

    Kei Zaitsu

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  1. Kazuaki Hisatsune
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  2. Tasuku Murata
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  3. Masaru Taniguchi
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  4. Tomomi Asano
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  5. Koretsugu Ogata
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  6. Akira Iguchi
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  7. Kei Zaitsu
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Contributions

Kazuaki Hisatsune: methodology, validation, formal analysis, investigation, data curation, writing — original draft, visualization. Tasuku Murata: methodology, validation, formal analysis, investigation, data curation. Masaru Taniguchi: investigation, writing — review and editing, visualization. Tomomi Asano: writing — review and editing, visualization. Koretsugu Ogata: methodology, validation, formal analysis, investigation, data curation. Akira Iguchi: data curation, writing — review and editing, visualization. Kei Zaitsu: conceptualization, methodology, writing — original draft, supervision, project administration, funding acquisition.

Corresponding authors

Correspondence to Kazuaki Hisatsune or Kei Zaitsu.

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Ethics approval

This work was approved by the Ethics Committee of Nagoya University Graduate School of Medicine (approval number: 2017-0175).

Source of biological material

Postmortem urine samples were obtained from three anonymous drug abusers by autopsy with the approval of the Ethics Committee of Nagoya University Graduate School of Medicine (approval number: 2017-0175).

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The authors declare are no competing interests.

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Hisatsune, K., Murata, T., Taniguchi, M. et al. Development of a rapid-fire drug screening method by probe electrospray ionization tandem mass spectrometry for human urine (RaDPi-U). Anal Bioanal Chem 416, 2503–2513 (2024). https://doi.org/10.1007/s00216-024-05215-x

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