Distribution and Visualisation of Chlorhexidine Within the Skin Using ToF-SIMS: A Potential Platform for the Design of More Efficacious Skin Antiseptic Formulations
In order to increase the efficacy of a topically applied antimicrobial compound the permeation profile, localisation and mechanism of action within the skin must first be investigated.
Time-of-flight secondary ion mass spectrometry (ToF-SIMS) was used to visualise the distribution of a conventional antimicrobial compound, chlorhexidine digluconate, within porcine skin without the need for laborious preparation, radio-labels or fluorescent tags.
High mass resolution and high spatial resolution mass spectra and chemical images were achieved when analysing chlorhexidine digluconate treated cryo-sectioned porcine skin sections by ToF-SIMS. The distribution of chlorhexidine digluconate was mapped throughout the skin sections and our studies indicate that the compound appears to be localised within the stratum corneum. In parallel, tape strips taken from chlorhexidine digluconate treated porcine skin were analysed by ToF-SIMS to support the distribution profile obtained from the skin sections.
ToF-SIMS can act as a powerful complementary technique to map the distribution of topically applied compounds within the skin.
KEY WORDSantisepsis chlorhexidine imaging mass spectrometry skin permeation ToF-SIMS
ACKNOWLEDGMENTS AND DISCLOSURES
This work was funded by an EPSRC industrial CASE award in collaboration with Dermal Technology Laboratory Ltd. The authors would like to acknowledge Professor Steve Chapman for his assistance and David Griffiths for his help and advice with the skin cryo-sectioning and histology. The authors state no conflict of interest.
- 2.Great Britain. Health Protection Agency, Quaterly epidermiological commentary: mandatory MRSA, MSSA, E. coli bacteraemia and C. difficile infection data (up to October–December 2011). London, UK; 2003.Google Scholar
- 6.Paulson SD. Handbook of topical antimicrobials: Industrial applications in consumer products. London: CRC press; 2003.Google Scholar
- 10.Williams AC. Transdermal and topical drug delivery from theory to clinical practice. London: Pharmaceutical Press; 2003.Google Scholar
- 11.Denton WG. Chlorhexidine. In: Block SS, editor. Sterilisation and preservation. 5th ed. London: Lippincott Williams and Wilkins; 1991. p. 274–89.Google Scholar
- 12.Davies D, Ward R, Heylings J. Multi-species assessment of electrical resistance as a skin integrity marker for in vitro percutaneous absorption studies. Toxicol In Vitro. 2004;18(3):351–8.Google Scholar
- 18.N’Dri-Stempfer B, Navidi WC, Guy RH, Bunge AL. Improved bioequivalence assessment of topical dermatological drug products using dermatopharmacokinetics. Pharm Res. 2009 Feb 2009;26(2):316–28.Google Scholar
- 22.Higo N, Naik A, Bommannan DB, Potts RO, Guy RH. Validation of reflectance infrared-spectroscopy as a quantitative method to measure percutaneous-absorption in-vivo. Pharm Res. 1993 Oct 1993;10(10):1500–6.Google Scholar
- 23.Dreher F, Modjtahedi BS, Modjtahedi SP, Maibach HI. Quantification of stratum corneum removal by adhesive tape stripping by total protein assay in 96-well microplates. Skin Res Technol. 2005 May 2005;11(2):97–101.Google Scholar
- 24.Lindemann U, Wilken K, Weigmann HJ, Schaefer H, Sterry W, Lademann J. Quantification of the horny layer using tape stripping and microscopic techniques. J Biomed Opt. 2003 Oct 2003;8(4):601–7.Google Scholar
- 25.Weigmann HJ, Lindemann U, Antoniou C, Tsikrikas GN, Stratigos AI, Katsambas A, et al. UV/VIS absorbance allows rapid, accurate, and reproducible mass determination of corneocytes removed by tape stripping. Skin Pharmacol Appl Skin Physiol. 2003 Jul–Aug 2003;16(4):217–27.Google Scholar
- 26.Voegeli R, Heiland J, Doppler S, Rawlings AV, Schreier T. Efficient and simple quantification of stratum corneum proteins on tape strippings by infrared densitometry. Skin Res Technol 2007 Aug 2007;13(3):242–51.Google Scholar
- 30.Saar BG, Contreras-Rojas LR, Xie XS, Guy RH. Imaging drug delivery to skin with stimulated raman scattering microscopy. Mol Pharm. 2011 May–Jun 2011;8(3):969–75.Google Scholar
- 32.Lademann J, Meinke MC, Schanzer S, Richter H, Darvin ME, Haag SF, et al. In vivo methods for the analysis of the penetration of topically applied substances in and through the skin barrier. Int J Cosmetic Sci. 2012;34(6):551–9.Google Scholar
- 36.Lagarrigue M, Becker M, Lavigne R, Deininger SO, Walch A, Aubry F, et al. Revisiting rat spermatogenesis with MALDI imaging at 20-μm resolution. Mol Cell Proteomics. 2011 doi: 10.1074/mcp.M110.005991.
- 41.SCCS, The Scientific Committee on Consumer Safety. Basic criteria for the in vitro assessment of dermal absorption of cosmetic ingredients. Updated, 2010; SCCS/1358/10.Google Scholar
- 48.Judd A, Scurr DJ, Heylings J, Wan KW, Griffiths D, Moss PG. Visualization of the permeation of chlorhexidine within the skin using Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS). In: Chilcott R, Brain KR, editors. Advances in dermatological science. London: Royal Society of Chemistry; 2013. In Press.Google Scholar