Skip to main content
SpringerLink
Log in

Localisation and quantification of benzalkonium chloride in eye tissue by TOF-SIMS imaging and liquid chromatography mass spectrometry

  • Research Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Benzalkonium (BAK) chloride is the most commonly used preservative in eye drops. It is generally composed of benzyldimethyldodecylammonium C12 and benzyldimethyltetradecylammonium C14 and is supposed to increase penetration of active compounds. However, numerous studies have reported its toxic effect to ocular surface especially in long-term treatments like against glaucoma, a sight-threatening disease. Albino rabbits were treated with a hyperosmolar solution and a high concentration of BAK solution for 1 month. Enucleated eyes were cryo-sectioned and analysed by mass spectrometry. Mass spectrometry imaging using time-of-flight secondary ion mass spectrometry (TOF-SIMS) has been used to characterize the spatial distribution and to determine the relative quantity of BAK at the surface of rabbit eye sections. Liquid chromatography coupled with mass spectrometry (LC-MS) using a hybrid linear ion trap-Orbitrap® mass spectrometer was used to obtain relative quantification of BAK at the sample surface. TOF-SIMS images of BAK ions indicated a distribution at the ocular surface and in deeper structures. Didecyldimethylammonium (DDMAC), which is used in hospitals as a substitute for BAK, was also detected and showed an accumulation around the eyes. After extraction with acetonitrile and chromatographic separation using a Gemini C18 column and an original elution gradient, the relative quantities of BAK and DDMAC present in the whole eye section surface were determined. This LC-MS method was validated in terms of limits of quantification, linearity, repeatability and reproducibility and its feasibility was evaluated in surgically obtained human samples. Specimens of iris, lens capsule or trabecular meshwork were found with significant levels of BAK and DDMAC, thus confirming the penetration of BAK in deep ocular structures, with potential deleterious effects induced by this cytotoxic compound. The analytical method developed here could therefore be of primary interest in the field of pharmaco-toxicology in order to localise, identify and quantify drugs or xenobiotic compounds present at biological sample surfaces.

Mass spectrometry image (TOF-SIMS) of rabbit eye conjunctiva treated with benzalkonium chloride

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

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Merianos JJ (1991) In: Block SS (ed) Disinfection, sterilization and preservation. Lea and Kebiger, Pittsburg, PA

    Google Scholar 

  2. Gomez-Gomar A, Gonzalez-Aubert MM, Garces-Torrents J, Costa-Segarra J (1990) J Pharm Biomed Anal 8:871–876

    Article  CAS  Google Scholar 

  3. Ambrus G, Takahashi LT, Marty PA (1987) J Pharm Sci 76:174–176

    Article  CAS  Google Scholar 

  4. Baudouin C, Labbe A, Liang H, Pauly A, Brignole-Baudouin F (2010) Progr Ret Eye Res 29:312–334

    Article  CAS  Google Scholar 

  5. Debbasch C, Brignole F, Pisella PJ, Warnet JM, Rat P, Baudouin C (2001) Invest Ophthalmol Vis Sci 42:642–652

    CAS  Google Scholar 

  6. Baudouin C (2008) Acta Ophthalomol 86:716–726

    Article  Google Scholar 

  7. Quigley HA (2011) Lancet 377:1367–1377

    Article  Google Scholar 

  8. Nùñez O, Moyano E, Galceran MT (2004) J Chromatogr A 1058:89–95

    Google Scholar 

  9. Fisher AA (1987) Cutis 39:381–383

    CAS  Google Scholar 

  10. Kuppens EV, De Jong CA, Stolwijk TR, De Kreizer RJ, Van Best JA (1995) Br J Ophthalmol 79:339–342

    Article  CAS  Google Scholar 

  11. Broadway DC, Grierson I, Hitchings RA (1993) Br J Ophthalmol 77:590–596

    Article  CAS  Google Scholar 

  12. Pól J, Strohalm M, Havlíček V, Volny M (2010) Histochem Cell Biol 34:423–434

    Article  Google Scholar 

  13. Garrett TJ, Menger RF, Dawson WW, Yost RA (2011) Anal Bioanal Chem 401:103–113

    Article  CAS  Google Scholar 

  14. Brignole-Baudouin F, Desbenoit N, Hamm G, Liang H, Both JP, Brunelle A, Fournier I, Guérineau V, Legouffe R, Stauber J, Touboul D, Wisztorski M, Salzet M, Laprévote O, Baudouin C (2012) PLoS One 7:e50180

    Article  CAS  Google Scholar 

  15. Touboul D, Laprévote O, Brunelle A (2011) Curr Opin Chem Biol 15:725–732

    Article  CAS  Google Scholar 

  16. Touboul D, Halgand F, Brunelle A, Kersting R, Tallarek E, Hagenhoff B, Laprévote O (2004) Anal Chem 76:1550–1559

    Article  CAS  Google Scholar 

  17. Sjövall P, Lausmaa J, Johansson B (2004) Anal Chem 76:4271–4278

    Article  Google Scholar 

  18. Touboul D, Kollmer F, Niehuis E, Brunelle A, Laprévote O (2005) J Am Soc Mass Spectrom 16:1608–1618

    Article  CAS  Google Scholar 

  19. Brunelle A, Laprévote O (2007) Curr Pharm Des 13:3335–3343

    Article  CAS  Google Scholar 

  20. Malmberg P, Jennische E, Nilsson D, Nygren H (2011) Anal Bioanal Chem 399:2711–2718

    Article  CAS  Google Scholar 

  21. Brunelle A, Laprévote O (2009) Anal Bioanal Chem 393:31–35

    Article  CAS  Google Scholar 

  22. Seyer A, Einhorn J, Brunelle A, Laprévote O (2010) Anal Chem 82:2326–2333

    Article  CAS  Google Scholar 

  23. Ferrer I, Furlong ET (2001) Environ Sci Technol 35:2583–2588

    Article  CAS  Google Scholar 

  24. Martinez-Carballo E, Sitka A, Gonzalez-Barreiro C, Kreuzinger N, Furhacker M, Scharf S, Gans O (2007) Environ Pollut 145:489–496

    Article  CAS  Google Scholar 

  25. Sutterlin H, Alexy R, Coker A, Kummerer K (2008) Chemosphere 72:479–484

    Article  CAS  Google Scholar 

  26. Van de Voorde A, Lorgeoux C, Gromaire MC, Chebbo G (2012) Environ Pollut 164:150–157

    Article  Google Scholar 

  27. Ferrer I, Furlong ET (2002) Anal Chem 74:1275–1280

    Article  CAS  Google Scholar 

  28. Li XL, Brownawell BJ (2009) Anal Chem 81:7926–7935

    Article  CAS  Google Scholar 

  29. Ganzera M, Aberham A, Stuppner H (2006) J Agr Food Chem 54:3768–3772

    Article  CAS  Google Scholar 

  30. Miyauchi T, Mori M, Ito K (2005) J Chromatogr A 1095:74–80

    Article  CAS  Google Scholar 

  31. Miyauchi T, Mori M, Ito K (2007) Holzforschung 61:337–341

    Article  CAS  Google Scholar 

  32. Acheampong AA, Small D, Baumgarten V, Welty D, Tang-Liu D (2002) J Ocul Pharmacol Th 18:325–337

    Article  CAS  Google Scholar 

  33. Makarov A, Denisov E, Kholomeev A, Balschun W, Lange O, Strupat K, Horning S (2006) Anal Chem 78:2113–2120

    Article  CAS  Google Scholar 

  34. Rubert J, James KJ, Manes J, Soler C (2012) J Chromatogr A 1223:84–92

    Article  CAS  Google Scholar 

  35. Pinhancos R, Maass S, Ramanathan DM (2011) J Mass Spectrom 46:1175–1181

    Article  CAS  Google Scholar 

  36. Gilmore IS, Seah MP (2002) Appl Surf Sci 187:89–100

    Article  CAS  Google Scholar 

  37. Sodhi RNS (2004) Analyst 129:483–487

    Article  CAS  Google Scholar 

  38. Brunelle A, Touboul D, Laprévote O (2005) J Mass Spectrom 40:985–999

    Article  CAS  Google Scholar 

  39. Vickerman JC (2001) In: J.C. Vickerman, D. Briggs D (eds) TOF-SIMS: surface analysis by mass spectrometry. SurfaceSpectra, Manchester

  40. Champeau E, Edelhauser H (1986) In: Holly F (ed) The preocular tear film. Dry Eye Institute, Inc, Lubbock, TX, pp 292–302

    Google Scholar 

Download references

Acknowledgements

This work and the post-doctoral fellowship position of ND were supported by the Agence Nationale de la Recherche (grant ANR-09-PIRI-0012 MASDA-EYE). Pascale Hamard (Quinze-Vingts National Hospital), Hong Liang (Quinze-Vingts National Hospital and INSERM) and Grégory Hamm (Imabiotech SAS) are gratefully acknowledged for their help in sample collection, experimental procedures and fruitful discussions during manuscript preparation, respectively.

Author information

Authors and Affiliations

  1. Centre de recherche de Gif, Institut de Chimie des Substances Naturelles, CNRS, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France

    Nicolas Desbenoit, Isabelle Schmitz-Afonso, David Touboul & Alain Brunelle

  2. INSERM, U968, Paris, 75012, France

    Nicolas Desbenoit, Christophe Baudouin & Françoise Brignole-Baudouin

  3. UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, 75012, France

    Nicolas Desbenoit, Christophe Baudouin & Françoise Brignole-Baudouin

  4. CNRS, UMR_7210, Paris, 75012, France

    Nicolas Desbenoit, Christophe Baudouin & Françoise Brignole-Baudouin

  5. Centre Hospitalier National d’Ophtalmologie des Quinze-Vingts, INSERM-DHOS CIC 503, Paris, 75012, France

    Christophe Baudouin & Françoise Brignole-Baudouin

  6. Université Versailles Saint-Quentin-en-Yvelines, Versailles, 78000, France

    Christophe Baudouin

  7. Assistance Publique-Hôpitaux de Paris Hôpital Ambroise Paré, Service d’Ophtalmologie, Boulogne-Billancourt, 92100, France

    Christophe Baudouin

  8. Faculté des sciences pharmaceutiques et biologiques, Université Paris Descartes, Sorbonne Paris Cité, Paris, France

    Olivier Laprévote & Françoise Brignole-Baudouin

  9. Assistance Publique-Hôpitaux de Paris Hôpital Lariboisière, Service de Toxicologie, 2 rue Ambroise Paré, 75010, Paris, France

    Olivier Laprévote

Authors
  1. Nicolas Desbenoit
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Isabelle Schmitz-Afonso
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Christophe Baudouin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Olivier Laprévote
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. David Touboul
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Françoise Brignole-Baudouin
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Alain Brunelle
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Isabelle Schmitz-Afonso.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Desbenoit, N., Schmitz-Afonso, I., Baudouin, C. et al. Localisation and quantification of benzalkonium chloride in eye tissue by TOF-SIMS imaging and liquid chromatography mass spectrometry. Anal Bioanal Chem 405, 4039–4049 (2013). https://doi.org/10.1007/s00216-013-6811-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-013-6811-7

Keywords

Search

Navigation