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Development of a Simple, Rapid, and Robust Isocratic Liquid Chromatographic Method for the Determination of Pyrimethamine and its Synthetic Impurities in Bulk Drugs and Pharmaceutical Formulations

Chromatographia volume 80, pages 1343–1352 (2017)Cite this article

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Abstract

Pyrimethamine is an important antiparasitic drug in the treatment of malaria and toxoplasmosis and is often used in combination with either sulfadoxine, sulfalene, or sulfadiazine. Determining the content of pyrimethamine and investigating the related substances is currently possible applying either a compendial monograph utilizing thin layer chromatography as well as liquid chromatographic methods used by the respective manufacturers. To provide a simple method which is capable of determining the content of pyrimethamine and of resolving four of its potential synthetic impurities a very simple, cheap, precise, and accurate isocratic RP-HPLC method was developed. All analytes can be separated within a total runtime of 30 min and the method was linear within the concentration ranges of 0.12–0.740, 0.104–0.621, 0.120–0.710, 2.0–11.8, and 1.01–5.80 µg mL−1 for pyrimethamine, impurity A, impurity B, impurity C, and impurity D, respectively. These substances were separated by employing a Eurospher-II C18H column (250 × 4.6 mm, 5 µm particle size), a mobile phase being a mixture of a 0.05 M KH2PO4 buffer solution (pH 2.6) and methanol in the ratio 40:60 (v/v). The analysis was carried out at 30 °C, applying a flow rate of 1.2 mL min−1, and a detection wavelength of λ = 215 nm. The coefficients of determinations (R 2) for the five analytes were greater than 0.994 for pyrimethamine and all impurities. Results of recovery studies were within the range of 89.1–105.1% for all substances. In all tested genuine batches of pyrimethamine raw material impurities within the specified limits were present which is concurrent with results obtained from using the present manufacturer’s method.

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Abbreviations

B. P.:

British Pharmacopoeia

EDQM:

The European Directorate for the Quality of Medicines and HealthCare

HPTLC:

High-performance thin layer chromatography

KAAD:

Catholic Academic Exchange Service

Ph. Eur.:

European Pharmacopoeia

Ph. Int.:

International Pharmacopoeia

RP-HPLC:

Reversed-phase high-performance liquid chromatography

TFDA:

Tanzania Food and Drugs Authority

TLC:

Thin layer chromatography

USP:

United States Pharmacopoeia

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Acknowledgements

Thanks to the KAAD (Catholic Academic Exchange Services, Bonn, Germany) for awarding scholarship to Yonah Hebron Mwalwisi, to the European Directorate for the Quality of Medicines and HealthCare (EDQM) for the provision of samples and reference substances, the Tanzania Food and Drugs Authority (Dar es salaam, Tanzania) for facilitating the research stay of Yonah H. Mwalwisi at the Institute of Pharmacy and Food Chemistry at Würzburg, and Christine Erk (Institute for Pharmacy and Food Chemistry, Würzburg) for LC/MS support.

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Authors and Affiliations

  1. Institute of Pharmacy and Food Chemistry, University of Würzburg, Am Hubland, 97074, Würzburg, Germany

    Yonah H. Mwalwisi, Ludwig Höllein & Ulrike Holzgrabe

  2. Tanzania Food and Drug Authority, Dar es Salaam, Tanzania

    Yonah H. Mwalwisi

  3. School of Pharmacy, Muhimbili University of Health and Allied Sciences, Dar es Salaam, Tanzania

    Eliangiringa Kaale

Authors
  1. Yonah H. Mwalwisi
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  2. Ludwig Höllein
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  3. Eliangiringa Kaale
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  4. Ulrike Holzgrabe
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Contributions

YHM, LH, and UH designed the study, YHM performed sample preparation and analysis, data collection, data analysis and interpretation. YHM and LH drafted the manuscript. UH and LH supervised the study and contributed in data interpretation. UH, LH, and EK reviewed of the manuscript. All authors have read and approved the final manuscript.

Corresponding author

Correspondence to Ulrike Holzgrabe.

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Conflict of interest

The authors declare that they have no competing interests.

Funding

This work was funded by the Catholic Academic Exchange Services, Bonn, Germany (KAAD) by providing a scholarship to YHM.

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Mwalwisi, Y.H., Höllein, L., Kaale, E. et al. Development of a Simple, Rapid, and Robust Isocratic Liquid Chromatographic Method for the Determination of Pyrimethamine and its Synthetic Impurities in Bulk Drugs and Pharmaceutical Formulations. Chromatographia 80, 1343–1352 (2017). https://doi.org/10.1007/s10337-017-3359-2

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  • DOI: https://doi.org/10.1007/s10337-017-3359-2

Keywords

  • Pyrimethamine
  • RP-HPLC
  • Quality control
  • Bulk drug and tablet analysis
  • Malaria
  • Impurity profiling