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Development and validation of an HPLC method for simultaneous determination of trimethoprim and sulfamethoxazole in human plasma

European Journal of Drug Metabolism and Pharmacokinetics volume 35, pages 41–46 (2010)Cite this article

Abstract

The combination of trimethoprim (TMP) and sulfamethoxazole (SMX) is used in the treatment of many common infections such as urinary, respiratory and gastrointestinal tract infections. The aim of this study was to determine TMP and SMX simultaneously in human plasma samples by high performance liquid chromatography (HPLC) using antipyrine as the internal standard. Separation of the compounds was achieved on a reverse-phase C8 column packed with 5 μm dimethyl octadecylsilyl bonded amorphous silica (4.6 mm × 250 mm) column using a mobile phase consisted of potassium hydrogen phosphate, acetonitrile, methanol and water adjusted to pH 6.2. The mobile phase was delivered at a flow rate of 1 mL min−1 and the effluent was monitored using Max plot technique at 25°C. Retention times were 5 min for TMP, 7 min for antipyrine and 9 min for SMX. Quantitation limits were 10 ng mL−1 for TMP and 50 ng mL−1 for SMX. Our findings indicated that the developed HPLC method was precise, accurate, specific and sensitive for simultaneous determination of TMP and SMX. Proposed HPLC method was successfully applied for the analysis of TMP and SMX in human plasma after oral administration of a co-trimoxazole tablet to human volunteers.

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Affiliations

  1. Faculty of Pharmacy, Hacettepe University, 06100, Ankara, Turkey

    Esin Sayar, Selma Sahin & A. Atilla Hıncal

  2. Ministry of Defense, Army Drug Factory, Ankara, Turkey

    Esin Sayar & Semsettin Cevheroglu

Authors
  1. Esin Sayar
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  2. Selma Sahin
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  3. Semsettin Cevheroglu
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  4. A. Atilla Hıncal
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Correspondence to Selma Sahin.

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Sayar, E., Sahin, S., Cevheroglu, S. et al. Development and validation of an HPLC method for simultaneous determination of trimethoprim and sulfamethoxazole in human plasma. Eur J Drug Metab Pharmacokinet 35, 41–46 (2010). https://doi.org/10.1007/s13318-010-0006-9

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  • DOI: https://doi.org/10.1007/s13318-010-0006-9

Keywords

  • Antipyrine
  • HPLC
  • Sulfamethoxazole
  • Trimethoprim
  • Co-trimoxazole