Archives of Toxicology

, Volume 80, Issue 10, pp 656–661 | Cite as

A dose-response study on the estrogenic activity of benzophenone-2 on various endpoints in the serum, pituitary and uterus of female rats

  • Christiane Schlecht
  • Holger Klammer
  • Wolfgang Wuttke
  • Hubertus JarryEmail author
Organ Toxicity and Mechanisms


The tetrahydroxylated biphenyl-ketone 2,2′,4,4′-tetrahydroxybenzophenone (BP2), one of twelve benzophenone-derived UV-filters, is used in cosmetic products and in packaging materials to protect these products from light induced damage. Recently published studies showed that BP2 exerts estrogenic activity; thus, it is an endocrine active chemical. We present data from a pharmacodynamic dose-response experiment with five dosages of BP2 applied per gavage to adult ovariectomized (ovx) rats for 5 days. Estradiol-valerate (E2) served as a control compound. The uterotrophic assay, proposed by the OECD, was modified to have a broader view on endocrine activity outside the urogenital tract to prevent that undesirable actions in other organs regulated by estrogens are missed. The gene expression levels of marker genes of estrogenic action were measured by semi-quantitative RT-PCR. Metabolic parameters were assessed by determination of the serum concentrations of leptin, cholesterol, high- and low-density lipoproteins, and triglycerides in the serum. Administration of BP2 at dosages of 10–1,000 mg/kg bodyweight led to changes of these parameters comparable to the changes in the E2 group with 0.6 mg/kg bodyweight. For the observed estrogenic activities of BP2, the “no observed adverse effect levels” were determined. Additionally, the data were further analyzed using the benchmark approach. If BP2 is transcutaneously absorbed in the human, the obtained threshold values would suggest refraining from the further use of BP2 as UV-filter in cosmetic products although additional toxicological studies should be conducted to clarify possible adverse effects.


2,2′,4,4′-tetrahydroxybenzophenone BP2 Multi-organic risk assessment Estradiol Estrogen receptor Benchmark approach Rat 







Complement protein 3


Truncated estrogen receptor protein 1


Estrogen receptor


Insulin-like growth factor 1


High/low density lipoprotein


Luteinizing hormone


Endocrine active chemical


No/lowest observed adverse effect level


Critical effect dose


Critical effect size


Confidence limit


Subunit of the hormones FSH, TSH and LH



This work was in part supported by the European Commission (EURISKED contract no. EVK1-CT2002-00128). The authors wish to thank Bas Bokkers from the IRAS (The Netherlands) for the help on the modeling process with the PROAST 02.R software. All experiments were performed according to the European Convention for the protection of vertebrate animals used for experimental and other scientific purposes (ETS 123) and GLP guidelines published by the OECD.


  1. Crump KS (1984) A new method for determining allowable daily intakes. Fundam Appl Toxicol 4:854–871CrossRefPubMedGoogle Scholar
  2. Crump KS (1995) Calculation of benchmark doses from continuous data. Risk Anal 15:79–89CrossRefGoogle Scholar
  3. Fasco MJ (1998) Estrogen receptor mRNA splice variants produced from the distal and proximal promoter transcripts. Mol Cell Endocrinol 138:51–59CrossRefPubMedGoogle Scholar
  4. Fattore E, Chu I, Sand S, Fanelli R, Falk-Filippson A, Hakansson H (2004) Dose-response assessment using the benchmark dose approach of changes in hepatic EROD activity for individual polychlorinated biphenyl congeners. Organohalogen Compounds 66:3402–3407Google Scholar
  5. Fernandez C, Nielloud F, Fortune R, Vian L, Marti-Mestres G (2002) Benzophenone-3: rapid prediction and evaluation using non-invasive methods of in vivo human penetration. J Pharm Biomed Anal 28:57–63CrossRefPubMedGoogle Scholar
  6. Friend KE, Ang LW, Shupnik MA (1995) Estrogen regulates the expression of several different estrogen receptor mRNA isoforms in rat pituitary. Proc Natl Acad Sci USA 92:4367–4371PubMedCrossRefGoogle Scholar
  7. Gupta VK, Zatz JL, Rerek M (1999) Percutaneous absorption of sunscreens through micro-yucatan pig skin in vitro. Pharm Res 16:1602–1607CrossRefPubMedGoogle Scholar
  8. Jarry H, Metten M, Spengler B, Christoffel V, Wuttke W (2003) In vitro effects of the Cimicifuga racemosa extract BNO 1055. Maturitas 44(Suppl 1):31–38CrossRefGoogle Scholar
  9. Jarry H, Christoffel J, Rimoldi G, Koch L, Wuttke W (2004) Multi-organic endocrine disrupting activity of the UV screen benzophenone 2 (BP2) in ovariectomized adult rats after 5 days treatment. Toxicology 205:87–93CrossRefPubMedGoogle Scholar
  10. Kalberlah F, Hassauer M (2003) Vergleich der Verfahren zur Ableitung gesundheitsbezogener Wirkungsschwellen (Benchmark - NOAEL). Abschlussbericht. Forschungs- und Beratungsinstitut Gefahrstoffe GmbH, Freiburg/BerlinGoogle Scholar
  11. KemI (2003) Human health risk assessment. Proposals for the use of assessment (uncertainty) factors. Application to risk assessment for plant protection products, industrial chemicals and biocidal products within the European Union. Body for Competence and Methodology Development, National Chemicals Inspectorate and Institute of Environmental Medicine, Karolinska Institutet, Solna, SwedenGoogle Scholar
  12. Kuiper GG, Carlsson B, Grandien K, Enmark E, Haggblad J, Nilsson S, Gustafsson JA (1997) Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors alpha and beta. Endocrinology 138:863–870CrossRefPubMedGoogle Scholar
  13. Miller D, Wheals BB, Beresford N, Sumpter JP (2001) Estrogenic activity of phenolic additives determined by an in vitro yeast bioassay. Environ Health Perspect 109:133–138PubMedCrossRefGoogle Scholar
  14. OECD (2001) Final Report of the Phase 1 of the Validation Study of the Uterotrophic Assay. Organisation for Economic Co-operation and Development, ParisGoogle Scholar
  15. Roth C, Leonhardt S, Seidel C, Luft H, Wuttke W, Jarry H (2000) Comparative analysis of different puberty inhibiting mechanisms of two GnRH agonists and the GnRH antagonist cetrorelix using a female rat model. Pediatr Res 48:468–474PubMedCrossRefGoogle Scholar
  16. SCCNFP/0321/00Final (2000). Notes of guidance for testing of cosmetic ingredients for their safty evaluation adopted by the SCCNFP on 24 October 2000Google Scholar
  17. Schlecht C, Klammer H, Jarry H, Wuttke W (2004) Effects of estradiol, benzophenone-2 and benzophenone-3 on the expression pattern of the estrogen receptors (ER) alpha and beta, the estrogen receptor-related receptor 1 (ERR1) and the aryl hydrocarbon receptor (AhR) in adult ovariectomized rats. Toxicology 205:123–130CrossRefPubMedGoogle Scholar
  18. Seidlova-Wuttke D, Hesse O, Jarry H, Christoffel V, Spengler B, Becker T, Wuttke W (2003) Evidence for selective estrogen receptor modulator activity in a black cohosh (Cimicifuga racemosa) extract: comparison with estradiol-17beta. Eur J Endocrinol 149:351–362CrossRefPubMedGoogle Scholar
  19. Seidlova-Wuttke D, Jarry H, Wuttke W (2004) Pure estrogenic effect of benzophenone-2 (BP2) but not of bisphenol A (BPA) and dibutylphtalate (DBP) in uterus, vagina and bone. Toxicology 205:103–112CrossRefPubMedGoogle Scholar
  20. Seidlova-Wuttke D, Jarry H, Christoffel J, Rimoldi G, Wuttke W (2005) Effects of bisphenol-A (BPA), dibutylphtalate (DBP), benzophenone-2 (BP2), procymidone (Proc), and linurone (Lin) on fat tissue, a variety of hormones and metabolic parameters: a 3 months comparison with effects of estradiol (E2) in ovariectomized (ovx) rats. Toxicology 213:13–24CrossRefPubMedGoogle Scholar
  21. Slob W (2002) Dose-response modeling of continuous endpoints. Toxicol Sci 66:298–312CrossRefPubMedGoogle Scholar
  22. Woutersen RA, Jonker D, Stevenson H, te Biesebeek JD, Slob W (2001) The benchmark approach applied to a 28-day toxicity study with Rhodorsil Silane in rats. The impact of increasing the number of dose groups. Food Chem Toxicol 39:697–707CrossRefPubMedGoogle Scholar
  23. Yamasaki K, Takeyoshi M, Yakabe Y, Sawaki M, Takatsuki M (2003) Comparison of the reporter gene assay for ER-alpha antagonists with the immature rat uterotrophic assay of 10 chemicals. Toxicol Lett 142:119–131CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Christiane Schlecht
    • 1
  • Holger Klammer
    • 1
  • Wolfgang Wuttke
    • 1
  • Hubertus Jarry
    • 1
    Email author
  1. 1.Department of Clinical and Experimental EndocrinologyUniversity of GoettingenGoettingenGermany

Personalised recommendations