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Characterization and cytotoxicity of a benzocaine inclusion complex

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Abstract

Benzocaine (BZC), is a local anesthetic widely used in topical formulations as well as in throat pastilles. A disadvantage is that the compound presents low aqueous solubility. The present work describes the preparation and characterization of an inclusion complex between BZC and β-cyclodextrin (β-CD), followed by cytotoxicity assays. The association constant (Ka) was calculated using solubility isotherms, at different temperatures, and an HPLC procedure, at room temperature, employing a reverse phase C18 column, with a mobile phase consisting of water/acetonitrile. Ka obtained with solubility isotherms at temperatures of 25, 35, and 45 °C were 229.8, 317.1, and 520.3 M−1, respectively. Employing HPLC, Ka was 38.0 M−1. The difference in the Ka value could be explained because HPLC analyses were conducted using organic solvent, which affected the host–guest interaction. Moreover, the continuous flow could have altered the degree of association of the drug with β-CD. The BZC/CD inclusion complex was characterized using infrared spectroscopy, thermogravimetry, and X-ray diffraction. Analysis showed a good agreement with literature, suggesting that the complex was established. Cytotoxicity assays using fibroblast V79 cells showed that BZC/CD formulation was not cytotoxic, demonstrating its potential to reduce the toxicity of the anesthetic. The assays demonstrated an effective interaction between BZC and CD, and that the inclusion complex was less toxic to V79 cells than the plain BZC, turning it a good alternative to decrease its toxicity when administered to patients.

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Acknowledgements

The authors thank Centro de Análises e Prospecção Química (CAPQ/UFLA), Departamento de Ciências do Solo (UFLA), and Laboratório de Cultura de Células (UNICAMP) for the provision of equipment and facilities. M.Z.C. received a fellowship from FAPEMIG, and L.M.A.P. received a fellowship from FAPESP.

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Authors and Affiliations

  1. Departamento de Química, UFLA – Universidade Federal de Lavras, Caixa Postal 3037, Lavras, Minas Gerais, CEP 37200-000, Brazil

    Liliane Henrique Torres, Mohana Zorkot de Carvalho, Adelir Aparecida Saczk & Luciana de Matos Alves Pinto

  2. Departamento de Bioquímica, Instituto de Biologia, UNICAMP – Universidade Estadual de Campinas, Campinas, São Paulo, Brazil

    Patrícia da Silva Melo & Eneida de Paula

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  1. Liliane Henrique Torres
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  2. Mohana Zorkot de Carvalho
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  3. Patrícia da Silva Melo
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  4. Eneida de Paula
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  5. Adelir Aparecida Saczk
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  6. Luciana de Matos Alves Pinto
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Correspondence to Luciana de Matos Alves Pinto.

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10847_2018_791_MOESM1_ESM.tif

Supplementary material 1 Decrease in BZC retention time in the presence of increasing concentrations of β-CD. Chromatographic conditions described in the methodology section (TIF 12 KB)

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Torres, L.H., de Carvalho, M.Z., Melo, P.d.S. et al. Characterization and cytotoxicity of a benzocaine inclusion complex. J Incl Phenom Macrocycl Chem 91, 9–15 (2018). https://doi.org/10.1007/s10847-018-0791-3

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  • DOI: https://doi.org/10.1007/s10847-018-0791-3

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