Skip to main content
SpringerLink
Log in

A New Colorimetric Assay for Studying and Rapid Screening of Membrane Penetration Enhancers¥

  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

Abstract

Purpose. This work aims to demonstrate a novel chemical assay for rapid screening and analysis of the mode of action of membrane interaction by penetration enhancers.

Methods. The new bio-mimetic membrane assembly, consisting of supramolecular aggregates of lipids and conjugated polydiacetylene, undergoes visible and quantifiable blue-red color transitions upon interaction with penetration enhancers.

Results. The new colorimetric model has been employed to examine various classes of penetration enhancers, including 1-dodecylhexahydro-2H-azepin-2-one (Azone), oleic acid, propylene-glycol, menthol, ethoxyglycol-diethyleneglycol-monoethyl-ether (Transcutol), polysorbate-polyethylenesorbitan-monolaurate (Tween-20), and the drug 7-chloro-1-methyl-5-phenyl-3H-1,4-benzodiazepin-2-one (Diazepam). The assay enables to evaluate the validity of various observations and hypotheses proposed in previous studies regarding permeation enhancement activities. Our results suggest, for example, that propylene glycol (PG) by itself does not interfere with membranes, but rather exhibits synergistic effect in combination with other penetration enhancers. Similarly, our data demonstrate that Transcutol does not independently interact with membranes. The colorimetric system also indicates that interaction of penetration enhancers with membranes depend upon the lipid phase, as well as the self-assembly properties of the enhancer molecules.

Conclusions. The new biomimetic model membrane system can be applied for rapid screening of the activities of penetration enhancers, and provides insight into the mechanisms of permeability of membrane-active compounds.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  1. H. Schaefer and T. E. Redelmeier. Skin Barrier. Principles of Percutaneous Absorption, Karger, Basel, 1996.

    Google Scholar 

  2. E. Wagner. Effects of membrane-active agents in gene delivery. J. Control. Release 53:155-158 (1998).

    Google Scholar 

  3. V. R. Goskonda, R. A. Hill, M. A. Khan, and I. K. Reddy. Permeability of chemical delivery systems across rabbit corneal (SIRC) cell line and isolated corneas: A comparative study. Pharm. Dev. Technol. 5:409-416 (2000).

    Google Scholar 

  4. A. F. El-Kattan, C. S. Asbill, and B. Michniak. The effect of terpene enhancer lipophilicity on the percutaneous permeation of hydrocortisone formulated in HPMC gel systems. Int. J. Pharm. 198:179-189 (2000).

    Google Scholar 

  5. E. Touitou, B. Godin, and C. Weiss. Enhanced Delivery of Drugs Into and Across the Skin by Ethosomal Carriers. Drug Devel. Res. 50:406-415 (2000).

    Google Scholar 

  6. Y. Kaplun-Frischoff and E. Touitou. Testosterone skin permeation enhancement by menthol through formation of eutectic with drug and interaction with skin lipids. J. Pharm. Sci 86:1394-1399 (1997).

    Google Scholar 

  7. C. L. Gay, T. M. Murphy, J. Hadgraft, I. W. Kellaway, J. C. Evans, and C. C. Rowlands. An electron spin resonance study of skin penetration enhancers. Int. J. Pharm. 49:39-45 (1989).

    Google Scholar 

  8. B. Ongpipattanakul, R. R. Burnette, R. O. Potts, and M. L. Francoeur. Evidence that oleic acid exists in a separate phase within stratum corneum lipids. Pharm. Res. 8:350-354 (1991).

    Google Scholar 

  9. B. W. Barry. Lipid protein partitioning theory of skin penetration enhancement. J. Control. Release 15:237-248 (1991).

    Google Scholar 

  10. S. Y. Okada, R. Jelinek, and D. H. Charych. Induced color change of conjugated polymeric vesicles by interfacial catalysis of phospholipase A-2. Angew. Chemie, Intl. Ed. Eng. 38:655-659 (1999).

    Google Scholar 

  11. S. Kolusheva, L. Boyer, and R. Jelinek. Colorimetric assay for rapid screening of anti-microbial peptides. Nature Biotechnology 18:225-227 (2000).

    Google Scholar 

  12. S. Kolusheva, T. Shahal, and R. Jelinek. Cation-selective color sensors composed of ionophore-phospholipid-polydiacetylene mixed vesicles. J. Am. Chem. Soc. 122:776-780 (2000).

    Google Scholar 

  13. S. Kolusheva, R. Kafri, M. Katz, and R. Jelinek. Rapid colorimetric detection of antibody-epitope recognition at a bio-mimetic membrane interface. J. Am. Chem. Soc. 123:417-422 (2001).

    Google Scholar 

  14. C. A. Philips, and B. Michniak. Transdermal delivery of drugs with differing lipophilicities using Azone analogs as dermal penetration enhancers. J. Pharm. Sci. 84:1427-1433 (1995).

    Google Scholar 

  15. E. Touitou and B. Fabin. Altered skin permeation of a highly lipophilic molecule. Int. J. Pharm. 43:9-15 (1988).

    Google Scholar 

  16. A. Naik, L. A. R. M. Pechtold, R. O. Potts, and R. H. Guy. Mechanism of oleic acid induced skin penetration enhancement in vivo in humans. J. Control. Release 37:299-306 (1995).

    Google Scholar 

  17. J. E. Harrison, A. C. Watkinson, D. M. Green, J. Hadgraft, and K. Brain. The relative effect of Azone and Transcutol on permeant diffusivity and solubility in human Stratum Corneum. Pharm. Res. 13:542-546 (1996).

    Google Scholar 

  18. J. R. Kunta, V. R. Goskonda, H. O. Brotherton, M. A. Khan, and I. K. Reddy. Effect of menthol and related terpenes on the percutaneous absorption of propanolol across excised hairless mouse skin. J. Pharm. Sci. 86:1369-1373 (1997).

    Google Scholar 

  19. J. A. Bouwstra, M. A. de Vries, G. S. Gooris, W. Bras, J. Brussee, and M. Ponec. Thermodynamic and structural aspects of the skin barrier. J. Control. Release 15:209-220 (1991).

    Google Scholar 

  20. P. P. Sarpotdar and J. L. Zatz. Percutaneous absorption enhancement by nonionic surfactants. Drug Develop. Indus. Pharm. 12:1625-1647 (1986).

    Google Scholar 

  21. D. H. Charych, J. O. Nagy, W. Spevak, and M. D. Bednarski. Direct colorimetric detection of a receptor-ligand interactions by a polymerized bilayer assembly Science 261:585-588 (1993).

    Google Scholar 

  22. H. Tanaka, M. A. Gomez, A. E. Tonelli, and M. Thakur. Thermochromic phase transitions of a polydiacetylene, poly(ETCD), studied by high resolution solid state C-13 NMR. Macromolecules 22:1208-1215 (1989).

    Google Scholar 

  23. R. R. Chance. Chromism in polydiacetylene solutions and crystals. Macromolecules 13:386-392 (1980).

    Google Scholar 

  24. G. M. Golden, J. E. McKie, and R. O. Potts. Role of stratum corneum lipid fluidity in transdermal drug flux. J. Pharm. Sci. 76:25-28 (1987).

    Google Scholar 

  25. N. V. Sheth, D. J. Freeman, W. I. Higuchi, and S. L. Spruance. The influence of Azone, propylene glycol and polyethylene glycol on in vitro skin penetration of trifluorothymidine. Int. J. Pharm. 28:201-209 (1986).

    Google Scholar 

  26. E. Touitou and L. Abed. Effect of propylene glycol, Azone and n-decylmethyl sulphoxide on skin permeation kinetics of 5-fluoroacil. Int. J. Pharm. 27:89-98 (1985).

    Google Scholar 

  27. T. M. Turunen, A. Urtti, P. Paronen, K. L. Audus, and J. H. Rytting. Effect of some penetration enhancers on epithelial membrane lipid domains: evidence from fluorescence spectroscopy studies. Pharm. Res. 11:288-294 (1994).

    Google Scholar 

  28. S. Y. Okada, S. Peng, W. Spevak, and D. H. Charych. Color and chromism of polydiacetylene vesicles. Accounts Chem. Res. 31:229-239 (1988).

    Google Scholar 

  29. Q. Cheng, M. Yamamoto, and R. C. Stevens. Amino-acid terminated polydiacteylene lipid microstrcutures: Morphology and chromatic transitions. Langmuir 16:5333-5342 (2000).

    Google Scholar 

  30. M. Shinitzky (Ed.) Biomembranes, Physical Aspects, John Wiley & Sons, New York, 1993.

    Google Scholar 

  31. W. A. Ritschel, R. Panchagnula, K. Stemmer, and M. Ashraf. Development of an intracutaneous depot for drugs. Binding, drug accumulation and retention studies, and mechanism of depot. Skin Pharmacol. 4:235-245 (1991).

    Google Scholar 

  32. E. Touitou, F. Levi-Schafer, N. Shaco-Ezra, R. Ben-Yossef, and B. Fabin. Enhanced permeation of theophylline through the skin and its effect on fibroblast proliferation. Int. J. Pharm. 70:159-166 (1991).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Chemistry and Stadler Minerva Center for Mesoscale Macromolecular Engineering, Ben Gurion University of the Negev, Beersheva, 84105, Israel

    Damien Evrard, Sofiya Kolusheva & Raz Jelinek

  2. Department of Pharmaceutics, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, 91120, Israel

    Elka Touitou

  3. Department of Life Science, Ben Gurion University of the Negev, Beersheva, Israel

    Yitzhak Fishov

Authors
  1. Damien Evrard
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Elka Touitou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sofiya Kolusheva
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yitzhak Fishov
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Raz Jelinek
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Evrard, D., Touitou, E., Kolusheva, S. et al. A New Colorimetric Assay for Studying and Rapid Screening of Membrane Penetration Enhancers¥. Pharm Res 18, 943–949 (2001). https://doi.org/10.1023/A:1010980009823

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1010980009823

Search

Navigation