In vitro skin absorption tests of three types of parabens using a Franz diffusion cell


The objective of this study was to evaluate the permeation of paraben derivatives — methylparaben (MP), propylparaben (PP), and butylparaben (BP) — in hairless mouse full skin and human cadaver epidermis using a Franz diffusion cell method, which is proposed as a reliable alternative method to an skin absorption test. Parabens, esterified hydroxybenzoic acid compounds, are widely used as preservatives in food, cosmetics, and pharmaceutical products. The skin permeation rate showed dose dependency, and the hairless mouse full skin showed a higher flux value than human cadaver epidermis. Among the permeability coefficient (Kp) values of three parabens, MP showed a higher Kp value than PP or BP. Hence, according to the definitions of Marzulli et al., parabens would be classified as “moderate” penetrants.

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  1. 1

    Soni MG, Taylor SL, Greenberg NA, Burdock GA . Evaluation of the health aspects of methyl paraben: a review of the published literature. Food Chem Toxicol 2002; 40: 1335–1373.

    CAS  Article  Google Scholar 

  2. 2

    Terasaki M, Kamata R, Shiraishi F, Makino M . Evaluation of estrogenic activity of parabens and their chlorinated derivatives by using the yeast two-hybrid assay and the enzyme-linked immunosorbent assay. Environ Toxicol Chem 2009; 28: 204–208.

    CAS  Article  Google Scholar 

  3. 3

    Giordano F, Bettini R, Donini C, Gazzaniga A, Caira MR, Zhang GG et al. Physical properties of parabens and their mixtures: solubility in water, thermal behavior, and crystal structures. J Pharm Sci 1999; 88: 1210–1216.

    CAS  Article  Google Scholar 

  4. 4

    Andersen FA . Final amended report on the safety assessment of methylparaben, ethylparaben, propylparaben, isopropylparaben, butylparaben, isobutylparaben, and benzylparaben as used in cosmetic products. Int J Toxicol 2008; 4: 1–82.

    Google Scholar 

  5. 5

    Calafat AM, Ye X, Wong LY, Bishop AM, Needham LL . Urinary concentrations of four parabens in the U.S. population: NHANES 2005-2006. Environ Health Perspect 2010; 118: 679–685.

    CAS  Article  Google Scholar 

  6. 6

    Elder RL . Final report on the safety assessment of methylparaben, ethylparaben, propylparaben and butylparaben. Int J Toxicol 1984; 3: 147–209.

    Google Scholar 

  7. 7

    Soni MG, Carabin IG, Burdock GA . Safety assessment of esters of p-hydroxybenzoic acid (parabens). Food Chem Toxicol 2005; 43: 985–1015.

    CAS  Article  Google Scholar 

  8. 8

    Barr L, Metaxas G, Harbach CAJ, Savoy LA, Darbre PD . Measurement of paraben concentrations in human breast tissue at serial locations across the breast form axilla to sternum. Toxicol Appl Pharmacol 2012; 32: 219–232.

    CAS  Google Scholar 

  9. 9

    National Toxicology Program 2005 NTP Concept Document. Butylparaben. Available at URL

  10. 10

    Blair RM, Fang H, Branham WS, Hass BS, Dial SL, Moland CL et al. The estrogen receptor binding affinities of 188 natural and xenochemicals: structural diversity of ligands. Toxicol Sci 2000; 54: 138–153.

    CAS  Article  Google Scholar 

  11. 11

    Darbre PD . Environmental oestrogens, cosmetics and breast cancer. Best Pract Res Clin Endocrinol Metab 2006; 20: 121–143.

    CAS  Article  Google Scholar 

  12. 12

    Darbre PD, Aljarrah A, Miller WR, Coldham NG, Sauer MJ, Pope GS . Concentrations of parabens in human breast tumours. J Appl Toxicol 2004; 24: 5–13.

    CAS  Article  Google Scholar 

  13. 13

    Darbre PD, Harvey PW . Paraben esters: review of recent studies of endocrine toxicity, absorption, esterase and human exposure, and discussion of potential human health risks. J Appl Toxicol 2008; 28: 561–578.

    CAS  Article  Google Scholar 

  14. 14

    Golden R, Gandy J, Vollmer G . A review of the endocrine activity of parabens and implications for potential risks to human health. Crit Rev Toxicol 2005; 35: 435–458.

    CAS  Article  Google Scholar 

  15. 15

    Harvey PW, Darbre P . Endocrine disrupters and human health: could oestrogenic chemicals in bodycare cosmetics adversely affect breast cancer incidence in women? A review of evidence and call for further research. J Appl Toxicol 2004; 24: 167–176.

    CAS  Article  Google Scholar 

  16. 16

    Spanier AJ, Fausnight T, Camacho TF, Braun JM . The associations of triclosan and paraben exposure with allergen sensitization and wheeze in children. Allergy Asthma Proc 2014; 35: 475–481.

    CAS  Article  Google Scholar 

  17. 17

    Towers CV, Terry PD, Lewis D, Howard B, Chambers W, Armistead C et al. Transplacental passage of antimicrobial paraben preservatives. J Expo Sci Environ Epidemiol 2015; 25: 604–607.

    CAS  Article  Google Scholar 

  18. 18

    Panel on Food Additives and Nutrient Sources Added to Food. Opinion of the scientific panel on food additives, flavourings, processing aids and materials in contact with food on a request from the commission related to para hydroxybenzoates (E 214-219), question number EFSA-Q-2004-063. EFSA J 2004; 83: 1–26.

    Google Scholar 

  19. 19

    Han F, He YZ, Yu CZ . On-line pretreatment and determination of parabens in cosmetic products by combination of flow injection analysis, solid-phase extraction and micellar electrokinetic chromatography. Talanta 2008; 74: 1371–1377.

    CAS  Article  Google Scholar 

  20. 20

    Hussein EI, Muret P, Berard M, Makki S, Humbert P . Assessment of principal parabens used in cosmetics after their passage through human epidermis-dermis layers (ex-vivo study). Exp Dermatol 2007; 16: 830–836.

    Article  Google Scholar 

  21. 21

    Bando H, Mohri S, Yamashita F, Takakura Y, Hashida M . Effects of skin metabolism on percutaneous penetration of lipophilic drugs. J Pharm Sci 1997; 86: 759–761.

    CAS  Article  Google Scholar 

  22. 22

    El Hussein S, Muret P, Berard M, Makki S, Humbert P . Assessment of principal parabens used in cosmetics after their passage through human epidermis-dermis layers (ex-vivo study). Exp Dermatol 2007; 16: 830–806.

    CAS  Article  Google Scholar 

  23. 23

    Brain KR, Walters KA, Green DM, Brain S, Loretz LJ, Sharma RK et al. Percutaneous penetration of diethanolamine through human skin in vitro: application from cosmetic vehicles. Food Chem Toxicol 2005; 43: 681–690.

    CAS  Article  Google Scholar 

  24. 24

    Mickelsen RL, Hall RC, Chern RT, Myers JR . Evaluation of a simple weight-loss method for determining the permeation of organic liquids through rubber films. Am Ind Hyg Assoc J 1991; 52: 445–447.

    CAS  Article  Google Scholar 

  25. 25

    Riviere JE, Brooks JD . Predicting skin permeability from complex chemical mixtures. Toxicol Appl Pharmacol 2005; 208: 99–110.

    CAS  Article  Google Scholar 

  26. 26

    OECD guidelines TG 428, Organization for Economic Cooperation and Development Guidance Document for the Conduct of Skin Absorption Studies OECD Series on Testing and Assessment, No. 28. OECD Publishing: Paris. 2004, 1–30.

  27. 27

    Nishizawa C, Takeshita K, Ueda J, Nakanishi I, Suzuki KT, Ozawa T . Reaction of para-hydroxybenzoic acid esters with singlet oxygen in the presence of glutathione produces glutathione conjugates of hydroquinone, potent inducers of oxidative stress. Free Radic Res 2006; 40: 233–240.

    CAS  Article  Google Scholar 

  28. 28

    Flynn GL, Dürrheim H, Higuchi WI . Permeation of hairless mouse skin II: membrane sectioning techniques and influence on alkanol permeabilities. J Pharm Sci 1981; 70: 52–56.

    CAS  Article  Google Scholar 

  29. 29

    Tadeuchi H, Mano Y, Terasaka S, Sakurai T, Furuya A, Urano H et al. Usefulness of rat skin as a substitute for human skin in the in vitro skin permeation study. Exp Anim 2011; 60: 373–384.

    Article  Google Scholar 

  30. 30

    Bartosova L, Bajgar J . Transdermal drug delivery in vitro using diffusion cells. Curr Med Chem 2012; 19: 4671–4677.

    CAS  Article  Google Scholar 

  31. 31

    Franz TJ . On the relevance of in vitro data. J Invest Dermatol 1975; 93: 633–640.

    Google Scholar 

  32. 32

    van de Sandt JJ, van Burgsteden JA, Cage S, Carmichael PL, Dick I, Kenyon S et al. In vitro predictions of skin absorption of caffeine, testosterone, and benzoic acid: a multi-centre comparison study. Regul Toxicol Pharmacol 2004; 39: 271–281.

    CAS  Article  Google Scholar 

  33. 33

    Venier M, Adami G, Larese F, Maina G, Renzi N . Percutaneous absorption of 5 glycol ethers through human skin in vitro. Toxicol In Vitro 2004; 18: 665–671.

    CAS  Article  Google Scholar 

  34. 34

    Reichling J, Landvatter U, Wagner H, Kostka KH, Schaefer UF . In vitro studies on release and human skin permeation of Australian tea tree oil (TTO) from topical formulations. Eur J Pharm Biopharm 2006; 64: 222–228.

    CAS  Article  Google Scholar 

  35. 35

    Marzulli FN, Brown DWC, Maibach HI . Techniques for studying skin penetration. Toxicol Appl Pharmacol 1969; 3: 76–83.

    Article  Google Scholar 

  36. 36

    Baert B, Boonen J, Burvenich C, Roche N, Stillaert F, Blondeel P et al. A new discriminative criterion for the development of franz diffusion tests for transdermal pharmaceuticals. J Pharm Pharm Sci 2010; 13: 218–230.

    CAS  Article  Google Scholar 

  37. 37

    Ng SF, Rouse JJ, Sanderson FD, Meidan V, Eccleston GM . Validation of a static franz diffusion cell system for in vitro permeation studies. AAPS PharmSci Tech 2010; 11: 1432–1441.

    Article  Google Scholar 

  38. 38

    Wróbel A, Gregoraszczuk EŁ . Effects of single and repeated in vitro exposure of three forms of parabens, methyl-, butyl- and propylparabens on the proliferation and estradiol secretion in MCF-7 and MCF-10A cells. Pharmacol Rep 2013; 65: 484–493.

    Article  Google Scholar 

  39. 39

    Franz TJ . Percutaneous absorption. On the relevance of in vitro data. J Invest Dermatol 1975; 64: 190–195.

    CAS  Article  Google Scholar 

  40. 40

    Takeuchi H, Ishida M, Furuya A, Todo H, Urano H, Sugibayashi K . Influence of skin thickness on the in vitro permeabilities of drugs through sprague-dawley rat or yucatan micropig skin. Biol Pharm Bull 2012; 35: 192–202.

    CAS  Article  Google Scholar 

  41. 41

    Netzlaff F, Kostka KH, Lehr CM, Schaefer UF . TEWL measurements as a routine method for evaluating the integrity of epidermis sheets in static Franz type diffusion cells in vitro. Limitations shown by transport data testing. Eur J Pharm Biopharm 2006; 63: 44–50.

    CAS  Article  Google Scholar 

  42. 42

    Guth K, Schäfer-Korting M, Fabian E, Landsiedel R, van Ravenzwaay B . Suitability of skin integrity tests for dermal absorption studies in vitro. Toxicol in Vitro 2015; 29: 113–123.

    CAS  Article  Google Scholar 

  43. 43

    Bond JR, Barry BW . Limitations of hairless mouse skin as a model for in vitro permeation studies through human skin: hydration damage. J Invest Dermatol 1988; 90: 486–489.

    CAS  Article  Google Scholar 

  44. 44

    Caon T, Costa AC, de Oliveira MA, Micke GA, Simões CM . Evaluation of the transdermal permeation of different paraben combinations through a pig ear skin model. Int J Pharm 2010; 391: 1–6.

    CAS  Article  Google Scholar 

  45. 45

    Cho WG . Comparison of drug delivery using hairless mouse and pig skin. J Korean Oil Chem Soc 2007; 24: 410–415.

    Google Scholar 

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This research was supported by grants (13162MFDS776) from Ministry of Food and Drug Safety in 2013.

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Correspondence to Bae-Hwan Kim.

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Seo, J., Kim, S. & Kim, B. In vitro skin absorption tests of three types of parabens using a Franz diffusion cell. J Expo Sci Environ Epidemiol 27, 320–325 (2017).

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  • Franz cell
  • in vitro
  • paraben
  • permeability coefficient
  • skin absorption

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