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An Improved UPLC Method for Rapid Analysis of Levofloxacin in Human Plasma

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Abstract

A rapid, specific, and sensitive ultra-performance liquid chromatographic method for analysis of levofloxacin in human plasma has been developed and validated. Plasma samples were spiked with the internal standard (enoxacin) and extracted with 10:1 (v/v) ethyl acetate–isopropanol. UPLC was performed on a 100 × 2.1 mm i.d., 1.7 µm particle, C18 column with 88:12 (v/v) 0.4% triethylamine buffer (pH 3)–acetonitrile as mobile phase, pumped isocratically at a pressure of 11,000 psi (758 bar) and a flow-rate of 0.3 mL min−1. Ultraviolet detection was performed at 300 nm. The retention times of levofloxacin and enoxacin were 3.4 and 2.8 min, respectively, and the run-time was 5 min. Calibration showed that response was a linear function of concentration over the range 0.05–10 µg mL−1 (r 2 ≥ 0.99) and the method was validated over this range for both precision and accuracy. The relative standard deviation was <15% for both intra-day and inter-day assay (n = 5). Levofloxacin and enoxacin were stable in plasma; there was no evidence of degradation during three freeze–thaw cycles, post-preparative stability at 20 °C was ≥24 h, short-term stability at room temperature was ≥6 h, and long-term stability at −70 °C was ≥30 days. The method was successfully used in a study of the bioequivalence of two levofloxacin tablet formulations in healthy volunteers.

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Acknowledgments

This research was supported by a grant from the Kyungpook National University Research Fund 2005 and the Korea Health 21 Research and Development Project, Ministry of Health and Welfare, Republic of South Korea (A050584) and by the Brain Korea 21 Project.

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Authors and Affiliations

  1. Clinical Trial Center, Kyungpook National University Hospital, 50 Samdok-2ga, Jung-gu, Daegu, 700-721, South Korea

    Dae-Jin Park, Prasad B. Phapale, Song Cui, Sung-Kyu Hwang & Young-Ran Yoon

  2. Department of Molecular Medicine, School of Medicine, Kyungpook National University, 101 Dongin-2ga, Jung-gu, Daegu, 700-422, South Korea

    Jung-Eun Kim & Young-Ran Yoon

  3. Department of Pharmacology and Clinical Pharmacology Unit, College of Medicine and Hospital, Seoul National University, 28 Yongon-dong, Jongno-gu, Seoul, South Korea

    In-Jin Jang

  4. Department of Pharmacology, College of Medicine, Kosin University, 34 Amnam-dong, Seo-gu, Busan, South Korea

    Dae-Jin Park & Woomi Kim

  5. Department of Biochemistry, Kyungpook National University, 1370 Sankyuk-dong, Buk-gu, Daegu, 702-701, South Korea

    Song Cui & Byung-Jo Moon

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  1. Dae-Jin Park
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Correspondence to Young-Ran Yoon.

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Park, DJ., Phapale, P.B., Jang, IJ. et al. An Improved UPLC Method for Rapid Analysis of Levofloxacin in Human Plasma. Chroma 68, 187–192 (2008). https://doi.org/10.1365/s10337-008-0669-4

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  • DOI: https://doi.org/10.1365/s10337-008-0669-4

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