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20.03.2019 | Original Article

A new method for high-resolution and high-precision analysis of flunitrazepam and 7-aminoflunitrazepam in human body fluids using a Monolithic SPE SpinTip and UPLC–Q-ToF–MS

verfasst von: Masaya Fujishiro, Ai Noguchi, Xiao-Pen Lee, Chika Hasegawa, Takeshi Kumazawa, Shota Miyazaki, Ayako Kuriki, Mari Hashimoto, Keizo Sato, Takaaki Matsuyama

Erschienen in: Forensic Toxicology | Ausgabe 2/2019

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Abstract

Purpose

Forensic toxicological analyses of drugs and their metabolites in human specimens usually require extractive pretreatment for successful analysis of substances from the matrix. In the present study, a high-throughput method was developed to analyze flunitrazepam, 7-aminoflunitrazepam, 7-acetamidoflunitrazepam, 7-acetamido-3-hydroxyflunitrazepam, and 3-hydroxyflunitrazepam in human plasma and urine samples using a new Monolithic C₁₈ gel-packed SpinTip and ultra-performance liquid chromatography (UPLC)–quadrupole time-of-flight (Q-ToF) mass spectrometry (MS).

Methods

Plasma (20 µL) or urine (100 µL) samples spiked with each component were extracted using a Monolithic C₁₈ SPE SpinTip and quantified by UPLC–Q-Tof–MS with positive-ion electrospray ionization (ESI).

Results

Good separation, with clear peak shapes of flunitrazepam and its metabolites, was achieved within an analysis time of 6 min, including the extraction time. Recoveries of flunitrazepam and 7-aminoflunitrazepam for plasma and urine samples were 93.5–118% and 97.7–109%, respectively. The regression equations for flunitrazepam and 7-aminoflunitrazepam showed excellent linearity in the range of 0.5–250 ng/mL for plasma and 0.4–500 ng/mL for urine, with detection limits of 0.2–0.5 ng/mL. Intra- and inter-day coefficients of variations for two drugs are smaller than 13.5%. The accuracy of quantitation was 89–110%.

Conclusions

The method was successfully applied to determine the level of flunitrazepam and its metabolites in human plasma and urine, respectively, after oral administration to a volunteer.

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Titel: A new method for high-resolution and high-precision analysis of flunitrazepam and 7-aminoflunitrazepam in human body fluids using a Monolithic SPE SpinTip and UPLC–Q-ToF–MS
verfasst von: Masaya Fujishiro
Ai Noguchi
Xiao-Pen Lee
Chika Hasegawa
Takeshi Kumazawa
Shota Miyazaki
Ayako Kuriki
Mari Hashimoto
Keizo Sato
Takaaki Matsuyama
Publikationsdatum: 20.03.2019
Verlag: Springer Singapore
Erschienen in: Forensic Toxicology / Ausgabe 2/2019
Print ISSN: 1860-8965
Elektronische ISSN: 1860-8973
DOI: https://doi.org/10.1007/s11419-019-00471-4

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A new method for high-resolution and high-precision analysis of flunitrazepam and 7-aminoflunitrazepam in human body fluids using a Monolithic SPE SpinTip and UPLC–Q-ToF–MS

Abstract

Purpose

Methods

Results

Conclusions

Neu im Fachgebiet Rechtsmedizin

Molekularpathologische Untersuchungen im Wandel der Zeit

Vergleichende Pathologie in der onkologischen Forschung

Gastrointestinale Stromatumoren

Personalisierte Medizin in der Onkologie

Live-Webinar "Chronische Unterbauch- und Viszeralschmerzen in der Praxis managen"

Springer Medizin

Abstract

Purpose

Methods

Results

Conclusions

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