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Application of micro-column HPLC to the determination of phenobarbital and carbamazepine in human blood serum

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Summary

The paper presents a method for the determination of phenobarbital and carbamazepine in human serum for routine application in therapy. Serum clean-up consists of the extraction of lipids with hexane, and the precipitation of proteins with lithium perchlorate—acetic acid—acetonitrile. The supernatant is subjected to HPLC on a micro-bore column with a C18 adsorbent with multi-wavelength detection at 210, 220, 230 and 240 nm. Gradient elution is performed with aqueous 0.2 M lithium perchlorate as solvent A and acetonitrile as solvent B. Compared to other methods, the solvent consumption is 10 times less. Multi-wavelength detection increases the reliability of peak identification. The respective detection limits for phenobarbital and carbamazepine are 0.1 μg and 0.05 μg per 1 mL of serum, with 50 μL of serum. The method makes it possible to determine other anticonvulsant preparations that are used to treat children and adults.

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Authors and Affiliations

  1. Limnological Institute of the Siberian Branch of the Russian Academy of Sciences, P.O. Box 4199, 664033, Irkutsk, Russia

    G. A. Fedorova, G. I. Baram & M. A. Grachev

  2. State Irkutsk District Children's Clinical Hospital, P.O. Box 4199, 664033, Irkutsk, Russia

    Yu. A. Aleksandrov & G. N. Tyuleneva

  3. Irkutsk Institute for Post-Graduate Training of Physicians, P.O. Box 4199, 664033, Irkutsk, Russia

    A. V. Starodubtsev

Authors
  1. G. A. Fedorova
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  2. G. I. Baram
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  3. M. A. Grachev
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  4. Yu. A. Aleksandrov
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  5. G. N. Tyuleneva
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  6. A. V. Starodubtsev
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Fedorova, G.A., Baram, G.I., Grachev, M.A. et al. Application of micro-column HPLC to the determination of phenobarbital and carbamazepine in human blood serum. Chromatographia 53, 495–497 (2001). https://doi.org/10.1007/BF02491610

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  • DOI: https://doi.org/10.1007/BF02491610

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