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Microextraction combined with microderivatization for drug monitoring and protein modification analysis from limited blood volume using mass spectrometry

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Abstract

In the clinic, ethosuximide is commonly used to treat generalized absence seizures but has recently been repurposed for other diseases. Because of adverse effects and drug interactions, high-throughput therapeutic drug monitoring of ethosuximide is necessary. Microextraction is a simple, effective, rapid, and low consumption of organic solvents method for sample preparation. In this study, microderivatization-increased detection (MDID)-combined microextraction was used to detect ethosuximide by mass spectrometry. Ethosuximide is a difficult to retain and ionize compound in the C18 nano-flow column and ionization interface, respectively. Hence, we developed a fast method for detecting ethosuximide in human plasma by using the MDID strategy (within 2 min). Chemical microderivatization parameters were studied and optimized to increase the sensitivity of ethosuximide detection at trace levels. The linear range for the analysis of ethosuximide in 10 μL plasma was 5–500 μg/mL with a coefficient of determination (r2) ≥ 0.995. The precision and accuracy of intraday and interday analyses of ethosuximide were below 13.0%. Furthermore, modifications of major proteins in plasma and blood cells, induced by ethosuximide, were identified. The proposed method effectively utilizes microliter samples to detect drug plasma concentrations under suitable microextraction procedures toward the eco-friendly goal of low consumption of organic solvents.

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Funding

This study was financially supported by grants from the Ministry of Science and Technology (MOST 106-2113-M-037-004 and 107-2113-M-037-003), the NSYSU-KMU Joint Research Project (NSYSUKMU107-P007), and the Research Center for Environmental Medicine, Kaohsiung Medical University. Instrumental support from the Center for Resources, Research and Development, Kaohsiung Medical University, is appreciated. This work was also partly supported by the Ministry of Education.

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Authors and Affiliations

  1. Department of Biochemistry, College of Medicine, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan

    Ying-Jung Wu, Yi-Shan Li & Chi-Yu Lu

  2. Department of Chemistry, College of Science, National Sun Yat-sen University, Kaohsiung, 80424, Taiwan

    Wei-Lung Tseng

  3. Research Center for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan

    Chi-Yu Lu

  4. Institute of Medical Science and Technology, National Sun Yat-sen University, Kaohsiung, 80424, Taiwan

    Chi-Yu Lu

  5. Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung, 80708, Taiwan

    Chi-Yu Lu

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  1. Ying-Jung Wu
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Correspondence to Chi-Yu Lu.

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The experiments were approved by the Institutional Review Board of Kaohsiung Medical University Chung-Ho Memorial Hospital. The method was performed in accordance with the approved guidelines and written informed consent was obtained from all participants.

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The authors declare that they have no conflict of interest.

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Wu, YJ., Li, YS., Tseng, WL. et al. Microextraction combined with microderivatization for drug monitoring and protein modification analysis from limited blood volume using mass spectrometry. Anal Bioanal Chem 410, 7405–7414 (2018). https://doi.org/10.1007/s00216-018-1349-3

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  • DOI: https://doi.org/10.1007/s00216-018-1349-3

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