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A rapid and simultaneous analysis of theophylline and its metabolites in human whole blood by ultra-performance liquid chromatography-tandem mass spectrometry

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Abstract

A rapid and sensitive method for analysis of theophylline and its six metabolites in human whole blood by ultra-performance liquid chromatography (UPLC)–tandem mass spectrometry (MS/MS) using 3-isobutyl-1-methylxanthine as internal standard is presented. After pretreatment of a blood sample by liquid–liquid extraction with acetonitrile, theophylline and its metabolites were analyzed by UPLC with a Poroshell 120 EC-C18 column and Acquity TQD tandem quadrupole mass spectrometer equipped with an electrospray ionization interface. The calibration curves for theophylline and its metabolites spiked into human blood showed equally good linearities in the microgram per milliliter range. The detection limits (signal-to-noise ratio 3) were as low as 0.1–1.0 ng/ml. The method gave satisfactory recovery rates, accuracy, and precision for quality control samples spiked with theophylline and its metabolites. To further validate the present method, 100 mg of theophylline was orally administered to a healthy male volunteer and the concentrations of theophylline and its metabolites in blood were measured 0.5, 1, 2, 4, and 8 h after dosing. The concentrations of theophylline in human blood were between 0.82 and 2.38 μg/ml. The present method should be very useful in forensic and clinical toxicology and pharmacokinetic studies.

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Acknowledgments

This work was supported by the Aikeikai Foundation.

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Authors and Affiliations

  1. Department of Legal Medicine, Aichi Medical University School of Medicine, Nagakute, Aichi, 480-1195, Japan

    Tadashi Ogawa, Hideki Hattori, Masae Iwai, Kenjiro Ito & Hiroshi Seno

  2. Department of Emergency and Critical Care Medicine, Aichi Medical University Graduate School of Medicine, Nagakute, Aichi, 480-1195, Japan

    Kenya Iwakura

  3. Department of Legal Medicine and Bioethics, Nagoya University Graduate School of Medicine, Nagoya, 466-8550, Japan

    Akira Ishii

  4. Hamamatsu University School of Medicine, Hamamatsu, 431-3192, Japan

    Osamu Suzuki

Authors
  1. Tadashi Ogawa
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  2. Hideki Hattori
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  3. Masae Iwai
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  4. Kenjiro Ito
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  5. Kenya Iwakura
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  6. Akira Ishii
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  7. Osamu Suzuki
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  8. Hiroshi Seno
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Correspondence to Tadashi Ogawa.

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Ogawa, T., Hattori, H., Iwai, M. et al. A rapid and simultaneous analysis of theophylline and its metabolites in human whole blood by ultra-performance liquid chromatography-tandem mass spectrometry. Forensic Toxicol 30, 142–148 (2012). https://doi.org/10.1007/s11419-012-0133-x

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  • DOI: https://doi.org/10.1007/s11419-012-0133-x

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