COMMENTS ON AN ANNEX XV DOSSIER FOR IDENTIFCATION OF A SUBSTANCE AS SVHC AND RESPONSES TO THESE COMMENTS

Substance name:	4-(1,1,3,3-tetramethylbutyl)phenol, (4-tert-Octylphenol)	
CAS number:	140-66-9	
EC number:	205-426-2	

The substance is proposed to be identified as meeting the following SVHC criteria set out in Article 57 of the REACH Regulation: 
EQUIVALENT LEVEL OF CONCERN (Art. 57(f))

Disclaimer: The European Chemicals Agency is not responsible for the content of this document. The Response to Comments table has been prepared by the competent authority of the Member State preparing the proposal for identification of a Substance of Very High Concern. The comments were received during the public consultation of the Annex XV dossier. RCOM has not been agreed by the Member State Committee nor has the document been modified as result of the MSC discussions.

PART I: Comments and responses to comments on the SVHC proposal and its justification

General comments on the SVHC proposal
Number	Date	Submitted by (name, Organisation/
MSCA)	Comment	Response	
27	2011/10/13	International NGO
Health and Environment Alliance
Belgium	HEAL supports the placement of Octylphenol on the Candidate List on the basis of the information in the Annex XV dossier, and commends Germany for initiating the first equivalent concern candidate nomination, particularly a chemical of endocrine disrupting properties.	Thank you for the support	
25	2011/10/13	Member State
Austria	Octylphenol is the first substance which is proposed as an SVHC because of its endocrine effects. As such effects pose a serious threat to the environment and are well described in the dossier, the Austrian CA can support the proposal. It is noted that most of the uses of octylphenol are intermediate uses and thus not covered by the authorisation regime. However, the report shows that free, unreacated octylphenol is present in measurable amounts in polymer matrices (partially as breakdown products) and this seems to be a major contamination source connected with the use of articles, e.g. articles made from phenol-formaldehyd resins.       	Thank you for the support and your comment

24	2011/10/13	National NGO
CHEM Trust
United Kingdom	CHEM Trust supports the inclusion of 4-(1,1,3,3-tetramethylbutyl)phenol also known as (4-tert-octylphenol) which is referred to as ‘octylphenol’ as a SVHC in the REACH candidate list based on the evidence presented that shows it to be a substance having endocrine disrupting properties.  However, CHEM Trust considers that it should be determined as having ED properties of relevance for both the environment and human health. 	Thank you for the support

The conclusion for 4-tert-octylphenol of being a substance of very high concern was drawn on the basis of environmental data. As neither the legal text nor the REACH guidance on SVHC identification states that this conclusion has to be drawn for the human health part as well to place a substance on the candidate list it is considered in accordance with the REACH regulation to propose 4-tert-octylphenol for the candidate list solely based on the conclusion for the environment.
The submission of the position paper is appreciated but it is not the intention of the dossier to anticipate the outcome on the criteria discussion for the assessment of substances that cause adverse effects mediated by endocrine mechanisms. 	
			See also:
Comment 24 attachment (CHEM Trust Position on EDC Criteria - Sept11.pdf).pdf		
23	2011/10/13	International NGO
WWF European Policy Office
Belgium	WWF supports the inclusion of 4-tert-octylphenol as SVHC of equivalent concern in the REACH candidate list based on its endocrine disrupting properties. However, we believe it should be determined as having ED properties of relevance for both the environment and human health.	Thank you for the support;
see response to COM 24	
			See also:
Comment 23 attachment (CHEM Trust Position on EDC Criteria - Sept11.pdf).pdf		
22	2011/10/13	National Authority
RIVM
Netherlands	The Netherlands supports the proposal to include 4-tert-octylphenol in the candidate list of substances of very high concern. 	Thank you for the support
	
20	2011/10/13	Industry or trade association
European Council for Alkylphenols and Derivatives (CEPAD)
Belgium	The REACH Annex XV Report on 4-(1,1,3,3-tetramethylbutyl)phenol (OP)   proposes classification of this compound as a Substance of Very High Concern (SVHC) on the basis of that it has “inherent  properties, which give rise to equivalent level of concern” to compounds classified as  CMR (Cat. 1A or 1B), PBT or vPvB  based on “environmental endocrine disrupting properties and subsequent serious effects to the environment”.  Article 57(f) states “substances giving rise to an equivalent level of concern” should have “scientific evidence of probable serious effects to human health or the environment”.  Therefore, categorization as a SVHC should be reserved for substances for which there is an adequate scientific basis to support a finding of effects that are both probable and serious.  The Annex XV Report for OP is in part based on invalid studies and flawed study interpretation and the overall assessment does not provide an adequate scientific basis to support a case that this compound rises to a level of concern that is equivalent to a CMR Category 1A, 1B, PBT or vPvB compound.

 As will be demonstrated in these comments, OP has only weak estrogenic activity that is 1000 to 40,000 fold less potent than 17ß-estradiol (E2) and 17α-ethynylestradiol (EE2), when calculated based on numerous reliable in vitro and in vivo studies.  Adverse apical effects caused by OP are not "clearly endocrine mediated", but rather are indicative of general toxicity possibly coupled with very weakly estrogenic activity.   The effects of OP are clearly not "comparable to other estrogens" as alleged in the Report.  

Results of aquatic toxicity tests conducted on OP and potent estrogens show that: 
1) effects on acute lethality, reproduction, growth, and development are very different for  the OP and EE2; 

2) studies with fish and frogs exposed to OP failed to fully link an endocrine mode of action affecting secondary endpoints with adverse reproductive, growth or developmental outcomes; and 

3) there are no definitive data to indicate that OP acts on the thyroid system of amphibians either as a thyroid agonist or antagonist. 

Following are detailed comments providing scientific evidence that support these statements as well as the conclusion that OP does not cause “probable serious effects” to the environment, nor is it of equivalent concern to CMR Cat. 1a and 1b, PBT, or vPvB substances.

Note: A complete comment document along with references and attachment (Tables A, B and C) are being submitted separately 

1.0.	The Annex XV Report on 4-(1,1,3,3-tetramethylbutyl)phenol (OP)  does not provide an adequate scientific basis to propose identifying this compound as a  Substance of Very High Concern (SVHC) on the basis of “equivalent concern” for endocrine modulated environmental effects; furthermore this proposal draws attention to the need for objective criteria  under REACH for what constitutes a  “probable serious” endocrine-mediated effect in aquatic species that are consistent with the CMR Cat. 1A and 1B toxicant definitions for mammalian species or PBT/vPvB classification criteria 

The REACH Annex XV Report for (OP) proposes classification of this compound as a SVHC “due to its inherent  properties which give rise to equivalent level of concern” to compounds classified as  CMR (Cat. 1A or 1B), PBT or vPvB  based on “environmental endocrine disrupting properties and subsequent serious effects to the environment”.

 The criteria for SVHC under Article 57(f) are: 

• Substances meeting the criteria for classification as carcinogenic, mutagenic or toxic for reproduction (CMR) category 1 or 2 in accordance with Directive 67/548/EEC; 

Note that CMR Cat. 1 and 2 under Directive 67/548/EEC are now categorized as “CMR Cat. 1A or 1B” under CLP EC Regulation No 1272/2008 (EC, 2008, Dec. 16) for Classification, Labelling and Packaging. As is the case in the Annex XV Report for OP, these comments will reflect these current CLP CMR category descriptions. 

• Substances which are persistent, bio-accumulative and toxic (PBT) in accordance with the criteria set out in Annex XIII of the REACH Regulation;

• Substances which are very persistent and very bio-accumulative (vPvB) in accordance with the criteria set out in Annex XIII of the REACH Regulation; and 

• Substances giving rise to an equivalent level of concern to substances meeting the above criteria. Such substances may have endocrine disrupting properties or have properties, that although not meeting the criteria for being a CMR, PBT or vPvB, there is scientific evidence of probable serious effects to human health or the environment. (Emphasis added)

1.1	Directive 67/548/EEC, and the more recent Classification, Labeling and Packaging Regulation No 1272/2008, provide criteria for classifying the certainty of the supporting data and the seriousness of effects in mammalian species, which allowed a clear assessment of the data for OP in the Annex XV Report that concluded that OP should not be classified as a CMR Cat. 1A or 1B. 
By virtue of the fact that there are criteria for defining serious reproductive effects (i.e., Toxic to Reproduction Cat. 1A or 1B), the German UVA was able to conduct an assessment of the reproductive effects of OP in mammals relative to those criteria and concluded “the adverse effects of 4-tert-octylphenol on female/male reproduction most probably related to its estrogen-active properties, were not considered appropriate to justify classification as toxic for reproduction and/or mutagenic / carcinogenic Category 1A or 1B”.  Category 1A relates to “known human reproductive toxicants” and Category 1B relates to “presumed human reproductive toxicants”.    It is important to also note that OP does not even meet the lesser criteria for Toxic to Reproduction Cat. 2, which relates to “suspected human reproductive toxicants” (Commission Regulation (EC) No 790/2009 First Amendment to CLP). 

Classification as Toxic to Reproduction Cat. 1A or 1B requires that “the data shall provide clear evidence of an adverse effect on sexual function and fertility or on development in the absence of other toxic effects or, if occurring together with other toxic effects, the adverse effect on reproduction is considered not to be a secondary non-specific consequence of other toxic effects” (EC, 2008, No. 1272/2008).    See Section 2.1 of these comments for further discussion on this point.  

1.2	Since reproductive effects in aquatic species are apical in nature and can be mediated by multiple modes of action, there is a need for criteria for determining the certainty and seriousness of  effects on reproduction in aquatic species for the purpose of defining SVHC compounds, just as there are criteria for mammalian species with the Reproduction Cat. 1A, 1B classifications.  

In its assessment of the mammalian effects of OP, the Annex XV Report acknowledges that multiple modes of action impact reproductive effects and, in the case of OP, modes of action other than endocrine may be more relevant. As discussed later in these comments, multiple modes of action also affect reproductive effects in aquatic species. In the case of OP, adverse apical reproductive effects occur in aquatic species only at exposure concentrations that are greater than the concentrations normally detected in the environment and where other toxic effects consistent with a narcotic mode of action also occur.  For a compound to be of “equivalent concern” to a Cat. 1A or 1B reproductive toxicant, it follows that reproductive effects in aquatic species should be subject to established criteria that are akin to those defined under R Cat. 1A or 1B for mammals. 

1.3	In the absence of established criteria, the Annex XV Report proposes criteria for assessing “equivalent concern” in the case of OP; however it does not provide adequate scientific evidence to support a conclusion that OP has probable serious effects in aquatic species that are of equivalent concern to CMR Cat. 1A or 1B, PBT or vPvB compounds.

Acknowledging the current lack of clear guidance for assessing “endocrine disruptors” and the lack of criteria defining “probable serious effects” in aquatic species that are “of equivalent concern” to other criteria for SVHCs, the Annex XV Report for OP proposes a basis for assessing this compound. 

The summary section (page 6) of Annex XV Report proposes that OP be identified as a “substance of equivalent concern” according to Article 57(f) based on the following: 

•	“Adverse effects observed in three fish species are clearly endocrine mediated according to the OECD draft guidance for endocrine disruptors (OECD, 2011)” and that OP is an endocrine disruptor based on these results;

•	“In all other tested fish species the endpoints affected are known to be influenced by estrogen activity and, with only one exemption, an estrogen mediated mode of action was observed. Population relevant endpoints affected in these species are known to be sensitive toward estrogen activity, although not being diagnostic for this mode of action”; and

•	“The effects observed in the amphibian species X.laevis after exposure to 4-tert-octylphenol are comparable to effects observed for 17 beta-estradiol. Effects that occurred in two other amphibian species (R.catesbetana and R. pipens) provide some evidence for a thyroid agonist mode of action. Both effect types indicate that 4-tert-octylphenol might be an endocrine disruptor in amphibians.” 

However, the Report does not provide adequate scientific evidence to support a conclusion that OP has probable serious endocrine effects in aquatic species that rise to the same level of concern as a CMR Cat. 1A or 1B, a PBT or a vPvB. 

The Report relies on the WHO-IPCS Global Assessment of Endocrine Disrupting Compounds for its definition of “endocrine disruptor”. The WHO-IPCS report recognizes that not all endocrine active compounds should be considered as endocrine disruptors.  Certainly, the potency, efficacy, as well as exposure concentrations, will affect the probability that a compound will actually cause endocrine disruption. In addition, the influence of other co-occurring modes of action will determine whether endocrine activity or disruption is the primary mode of action. 

As will be discussed further in these comments, a consistent and treatment-related range of NOEC and LOEC values (10 to 900 μg/L) have been reported in studies on OP  that addressed a range of relevant apical endpoints (i.e.,  survival, growth and development, and reproduction)  in a range of species and taxa (i.e., fish, amphibians and invertebrates) covering all parts of the test organisms’ life cycles (from eggs to reproducing adults) including life stages likely to be sensitive to an endocrine mode of action. Effects reported for endocrine sensitive endpoints occur at concentrations within the same range of NOECs and LOECs that are also consistent with a narcotic mode of action.  It is also important to note that concentrations of OP observed in the environment are generally at least an order of magnitude less than the lowest value in this range of LOEC and NOECs.

The remainder of these comments addresses the data and the assessment for OP provided in the Annex XV Report, which does not support a conclusion of “equivalent concern” to CMR Cat. 1A, 1B or PBT or vPvB chemicals.  These comments raise the need for ECHA to develop criteria for determining probable and serious effects in aquatic species  Endocrine mediated reproductive effects that are “known” or “presumed” based on supporting data that show they occur in the absence of other interfering toxicological mechanisms may rise to a similar level concern as Cat. 1A or 1B reproductive toxicant, particularly when effects occur at exposures that are environmentally relevant. However, reproductive effects that occur at only concentrations that are so high that other modes of toxicity are also occurring or that are not environmentally relevant should not be considered to be of equivalent concern to CMR Cat. 1A, 1B or PBT or vPvB compounds. 
	Thank you for your extensive comment. Please find below our reply to some of the aspect mentioned. Further aspects are considered in our responses to your specific comments.
Please be assured that your points were thoroughly analysed and some details in the dossier were changed based on your comments. As described in detail in our responses to your specific comments, aspects changed were not decisive for the overall conclusion. The conclusion that 4-tert-octylphenol should be identified as substance of very high concern due to its endocrine disrupting adverse effects in aquatic organisms still remains valid. 

With regard to point 1) – 3) please see our response to your specific comments 

1.0 We appreciate your description of Art 57 criteria and agree that objective criteria would facilitate SVHC identification for endocrine disruptors. However we believe that they are not a prerequisite for a case by case decision as indicated in Art 57 (f)

We assume that you are referring to Art 57 (not Art 57(f)) in the following paragraphs? 

1.1 Thanks for the comments. They are appreciated and will provide valuable input in further discussions.

1.2 We appreciate your comments about criteria for reproductive effects in wildlife. However you might b aware, that the proposal is based on the fact that OP results in severe endocrine disrupting apical effects in aquatic organisms and that due to this fact the substance raises equivalent concern compared to PBT/vPvB and CMR substances. This proposal is not based on the reproductive effects as such.
Criteria for reproductive effects are not considered necessary for this purpose.

1.3 In order to avoid misunderstanding we would like to emphasize that the dossier did not aim at proposing any criteria. 
With regard to your comment, that no adequate scientific evidence is provided for the conclusion that OP should be regarded as SVHC, please consider our responses to comments 6.1 and 6.2.

Please note that the WHO definition does not consider potency and exposure concentrations nor does it require the endocrine mode of action to be the primary mode of action.

Thank you for acknowledging the consistent data set and the fact that clearly endocrine mediated apical endpoints are among the most sensitive endpoints. Please see our responses to comments 6.0 ff with regard to the question whether or not adverse effects observed are endocrine mediated or not. 

Thank you for your comment with regard to criteria that should be used to identify SVHCs based on their endocrine disrupting apical effects. Please note that the dossier does not aim at pre-empting this discussion.
	
			See also:
Comment 20 attachment (CEPAD-APERC Comments on OP SVHC (Oct. 13, 2011).zip).zip		
19	2011/10/13	Industry or trade association
Alkylphenols & Ethoxylates Research Council
United States	The REACH Annex XV Report on 4-(1,1,3,3-tetramethylbutyl)phenol (OP)   proposes classification of this compound as a Substance of Very High Concern (SVHC) on the basis of that it has “inherent  properties, which give rise to equivalent level of concern” to compounds classified as  CMR (Cat. 1A or 1B), PBT or vPvB  based on “environmental endocrine disrupting properties and subsequent serious effects to the environment”.  Article 57(f) states “substances giving rise to an equivalent level of concern” should have “scientific evidence of probable serious effects to human health or the environment”.  Therefore, categorization as a SVHC should be reserved for substances for which there is an adequate scientific basis to support a finding of effects that are both probable and serious.  The Annex XV Report for OP is in part based on invalid studies and flawed study interpretation and the overall assessment does not provide an adequate scientific basis to support a case that this compound rises to a level of concern that is equivalent to a CMR Category 1A, 1B, PBT or vPvB compound.

 As will be demonstrated in these comments, OP has only weak estrogenic activity that is 1000 to 40,000 fold less potent than 17ß-estradiol (E2) and 17α-ethynylestradiol (EE2), when calculated based on numerous reliable in vitro and in vivo studies.  Adverse apical effects caused by OP are not "clearly endocrine mediated", but rather are indicative of general toxicity possibly coupled with very weakly estrogenic activity.   The effects of OP are clearly not "comparable to other estrogens" as alleged in the Report.  

Results of aquatic toxicity tests conducted on OP and potent estrogens show that: 
1) effects on acute lethality, reproduction, growth, and development are very different for        the OP and EE2; 
2) studies with fish and frogs exposed to OP failed to fully link an endocrine mode of action affecting secondary endpoints with adverse reproductive, growth or developmental outcomes; and 
3) there are no definitive data to indicate that OP acts on the thyroid system of amphibians either as a thyroid agonist or antagonist. 

Following are detailed comments providing scientific evidence that support these statements as well as the conclusion that OP does not cause “probable serious effects” to the environment, nor is it of equivalent concern to CMR Cat. 1a and 1b, PBT, or vPvB substances.

Note: A complete comment document along with references and attachment (Tables A, B and C) are being submitted separately 

1.0.	The Annex XV Report on 4-(1,1,3,3-tetramethylbutyl)phenol (OP)  does not provide an adequate scientific basis to propose identifying this compound as a  Substance of Very High Concern (SVHC) on the basis of “equivalent concern” for endocrine modulated environmental effects; furthermore this proposal draws attention to the need for objective criteria  under REACH for what constitutes a  “probable serious” endocrine-mediated effect in aquatic species that are consistent with the CMR Cat. 1A and 1B toxicant definitions for mammalian species or PBT/vPvB classification criteria 

The REACH Annex XV Report for (OP) proposes classification of this compound as a SVHC “due to its inherent  properties which give rise to equivalent level of concern” to compounds classified as  CMR (Cat. 1A or 1B), PBT or vPvB  based on “environmental endocrine disrupting properties and subsequent serious effects to the environment”.

 The criteria for SVHC under Article 57(f) are: 

• Substances meeting the criteria for classification as carcinogenic, mutagenic or toxic for reproduction (CMR) category 1 or 2 in accordance with Directive 67/548/EEC; 

Note that CMR Cat. 1 and 2 under Directive 67/548/EEC are now categorized as “CMR Cat. 1A or 1B” under CLP EC Regulation No 1272/2008 (EC, 2008, Dec. 16) for Classification, Labelling and Packaging. As is the case in the Annex XV Report for OP, these comments will reflect these current CLP CMR category descriptions. 

• Substances which are persistent, bio-accumulative and toxic (PBT) in accordance with the criteria set out in Annex XIII of the REACH Regulation;

• Substances which are very persistent and very bio-accumulative (vPvB) in accordance with the criteria set out in Annex XIII of the REACH Regulation; and 

• Substances giving rise to an equivalent level of concern to substances meeting the above criteria. Such substances may have endocrine disrupting properties or have properties, that although not meeting the criteria for being a CMR, PBT or vPvB, there is scientific evidence of probable serious effects to human health or the environment. (Emphasis added)

1.1	Directive 67/548/EEC, and the more recent Classification, Labeling and Packaging Regulation No 1272/2008, provide criteria for classifying the certainty of the supporting data and the seriousness of effects in mammalian species, which allowed a clear assessment of the data for OP in the Annex XV Report that concluded that OP should not be classified as a CMR Cat. 1A or 1B. 
By virtue of the fact that there are criteria for defining serious reproductive effects (i.e., Toxic to Reproduction Cat. 1A or 1B), the German UVA was able to conduct an assessment of the reproductive effects of OP in mammals relative to those criteria and concluded “the adverse effects of 4-tert-octylphenol on female/male reproduction most probably related to its estrogen-active properties, were not considered appropriate to justify classification as toxic for reproduction and/or mutagenic / carcinogenic Category 1A or 1B”.  Category 1A relates to “known human reproductive toxicants” and Category 1B relates to “presumed human reproductive toxicants”.    It is important to also note that OP does not even meet the lesser criteria for Toxic to Reproduction Cat. 2, which relates to “suspected human reproductive toxicants” (Commission Regulation (EC) No 790/2009 First Amendment to CLP). 

Classification as Toxic to Reproduction Cat. 1A or 1B requires that “the data shall provide clear evidence of an adverse effect on sexual function and fertility or on development in the absence of other toxic effects or, if occurring together with other toxic effects, the adverse effect on reproduction is considered not to be a secondary non-specific consequence of other toxic effects” (EC, 2008, No. 272/2008).    See Section 2.1 of these comments for further discussion on this point.  

1.2	Since reproductive effects in aquatic species are apical in nature and can be mediated by multiple modes of action, there is a need for criteria for determining the certainty and seriousness of  effects on reproduction in aquatic species for the purpose of defining SVHC compounds, just as there are criteria for mammalian species with the Reproduction Cat. 1A, 1B classifications.  

In its assessment of the mammalian effects of OP, the Annex XV Report acknowledges that multiple modes of action impact reproductive effects and, in the case of OP, modes of action other than endocrine may be more relevant. As discussed later in these comments, multiple modes of action also affect reproductive effects in aquatic species. In the case of OP, adverse apical reproductive effects occur in aquatic species only at exposure concentrations that are greater than the concentrations normally detected in the environment and where other toxic effects consistent with a narcotic mode of action also occur.  For a compound to be of “equivalent concern” to a Cat. 1A or 1B reproductive toxicant, it follows that reproductive effects in aquatic species should be subject to established criteria that are akin to those defined under R Cat. 1A or 1B for mammals. 

1.3	In the absence of established criteria, the Annex XV Report proposes criteria for assessing “equivalent concern” in the case of OP; however it does not provide adequate scientific evidence to support a conclusion that OP has probable serious effects in aquatic species that are of equivalent concern to CMR Cat. 1A or 1B, PBT or vPvB compounds.

Acknowledging the current lack of clear guidance for assessing “endocrine disruptors” and the lack of criteria defining “probable serious effects” in aquatic species that are “of equivalent concern” to other criteria for SVHCs, the Annex XV Report for OP proposes a basis for assessing this compound. 

The summary section (page 6) of Annex XV Report proposes that OP be identified as a “substance of equivalent concern” according to Article 57(f) based on the following: 

•	“Adverse effects observed in three fish species are clearly endocrine mediated according to the OECD draft guidance for endocrine disruptors (OECD, 2011)” and that OP is an endocrine disruptor based on these results;

•	“In all other tested fish species the endpoints affected are known to be influenced by estrogen activity and, with only one exemption, an estrogen mediated mode of action was observed. Population relevant endpoints affected in these species are known to be sensitive toward estrogen activity, although not being diagnostic for this mode of action”; and

•	“The effects observed in the amphibian species X.laevis after exposure to 4-tert-octylphenol are comparable to effects observed for 17 beta-estradiol. Effects that occurred in two other amphibian species (R.catesbetana and R. pipens) provide some evidence for a thyroid agonist mode of action. Both effect types indicate that 4-tert-octylphenol might be an endocrine disruptor in amphibians.” 

However, the Report does not provide adequate scientific evidence to support a conclusion that OP has probable serious endocrine effects in aquatic species that rise to the same level of concern as a CMR Cat. 1A or 1B, a PBT or a vPvB. 

The Report relies on the WHO-IPCS Global Assessment of Endocrine Disrupting Compounds for its definition of “endocrine disruptor”. The WHO-IPCS report recognizes that not all endocrine active compounds should be considered as endocrine disruptors.  Certainly, the potency, efficacy, as well as exposure concentrations, will affect the probability that a compound will actually cause endocrine disruption. In addition, the influence of other co-occurring modes of action will determine whether endocrine activity or disruption is the primary mode of action.  As will be discussed further in these comments, a consistent and treatment-related range of NOEC and LOEC values (10 to 900 μg/L) have been reported in studies on OP  that addressed a range of relevant apical endpoints (i.e.,  survival, growth and development, and reproduction)  in a range of species and taxa (i.e., fish, amphibians and invertebrates) covering all parts of the test organisms’ life cycles (from eggs to reproducing adults) including life stages likely to be sensitive to an endocrine mode of action. Effects reported for endocrine sensitive endpoints occur at concentrations within the same range of NOECs and LOECs that are also consistent with a narcotic mode of action.  It is also important to note that concentrations of OP observed in the environment are generally at least an order of magnitude less than the lowest value in this range of LOEC and NOECs.

The remainder of these comments addresses the data and the assessment for OP provided in the Annex XV Report, which does not support a conclusion of “equivalent concern” to CMR Cat. 1A, 1B or PBT or vPvB chemicals.  These comments raise the need for ECHA to develop criteria for determining probable and serious effects in aquatic species  Endocrine mediated reproductive effects that are “known” or “presumed” based on supporting data that show they occur in the absence of other interfering toxicological mechanisms may rise to a similar level concern as Cat. 1A or 1B reproductive toxicant, particularly when effects occur at exposures that are environmentally relevant. However, reproductive effects that occur at only concentrations that are so high that other modes of toxicity are also occurring or that are not environmentally relevant should not be considered to be of equivalent concern to CMR Cat. 1A, 1B or PBT or vPvB compounds. 	See response to comment 20	
			See also:
Comment 19 attachment (CEPAD-APERC Comments on OP SVHC (Oct. 13, 2011).zip).zip		
18	2011/10/13	National NGO
Danish Ecological Council
Denmark	Overall, the Danish Ecological Council support the Annex XV dossier, and we agree that 4-tert-octylphenol is an endocrine disruptor and should be included on the candidate list as a SVHC of an equivalent level of concern (article 57f). However, we do not agree that 4-tert-octylphenol is only an endocrine disrupter in the aspect of ecotoxicology but believe that it is also of concern for human health, which is amplified by its occurrence in wide spread applications, whereas many are close to consumers. Therefore we argue, that the conclusion should be amended.
There are new data for endocrine disrupting effects of the substance from different test systems and species (e.g. in vitro studies, fish, and rodents) that indicate that 4-tert-octylphenol acts as an endocrine disrupter mainly through the estrogen receptor, which is present in a wide variety of species, including humans.
In conclusion, we argue that both environmental and human health concerns should be reflected in the conclusion of the dossier regardless of whether the evidence originates from ecotoxicity or toxicity studies. Since these considerations might have an impact on further considerations regarding possible Authorizations, it is very important that they get generally accepted. It will furthermore influence the future possibility to regulate combined exposure to endocrine disrupters.	Thank you for the support

see response to COM 24	
17	2011/10/13	National NGO
Réseau Environnement Santé
France	Our organisation, RES, supports the Annex XV dossier on 4-tert Octylphenol on the basis of endocrine disrupting properties of this chemical. Yet, we think the dossier undermines the risks for human health. The widespread presence of OP and human exposure are well documented. Recent research suggests that OP has potential for endocrine disruption in mammals, as a single substance and/or in mixture with other ubiquitous chemicals. 	Thank you for the support

see response to COM 24	
			See also:
Comment 17 attachment (Annex RES octylphenol.pdf).pdf		
16	2011/10/13	Member State
United Kingdom	1) The Commission’s final decision on the definition of an endocrine disrupting substance is not expected until 2013.  As an opening remark, we think it would be highly damaging for the credibility of the regulatory process if the approach adopted in this dossier, should it be accepted, were to fall outside the scope of the definition in due course. We therefore think it would have been preferable to wait for the definition to become available before submitting the dossier. The intervening time could then have been used to gather better exposure information. However, now that the dossier has been tabled, we are obliged to pass comment on it.

2) The German c.a. is to be congratulated at the comprehensiveness of their approach to the description of endocrine effects for this substance in wildlife. However, in producing summaries at various places in the text, the limited reliability of some of the studies is sometimes forgotten. For example, all of the mollusc studies receive a Klimisch score of 3, yet it is stated that “4-tert-octylphenol is considered to be an endocrine disrupter causing adverse effects in... molluscs” (page 6). We do not believe that such statements should be made.

Thank you for your comment. The summaries were checked. Low quality data was clearly marked and summaries were rephrased in order to avoid misunderstanding. 	
14	2011/10/13	International NGO
ChemSec
Sweden	In short we support the Annex XV dossier as a whole and the conclusion that octylphenol is to be considered an endocrine disruptor but we have several remarks with regard to the human health side where the conclusion is not satisfactory protective of human health due to the reasons mentioned below;

Taking into consideration the below mentioned new data on endocrine disrupting effects in mammals, the lack data on probable exposure routes and the very high production volume of octylphenol and its occurrence in wide spread applications whereof many close to consumers, the conclusion that should be reached is that octylphenol is indeed an endocrine disruptor also in mammals.	Thank you for the support

see response to COM 24	
			See also:
Comment 14 attachment (Annex A_references_Octylphenol.doc).doc		
13	2011/10/13	National NGO
Swedish Society for Nature Conservation (SSNC)
Sweden	In short we support the Annex XV dossier as a whole and the conclusion that octylphenol is to be considered an endocrine disruptor but we have several remarks with regard to the human health side where the conclusion is not satisfactory protective of human health due to the reasons mentioned below;

Taking into consideration the below mentioned new data on endocrine disrupting effects in mammals, the lack data on probable exposure routes and the very high production volume of octylphenol and its occurrence in wide spread applications whereof many close to consumers, the conclusion that should be reached is that octylphenol is indeed an endocrine disruptor also in mammals.	Thank you for the support

see response to COM 24	
12	2011/10/13	International NGO
ClientEarth
Belgium	ClientEarth support the Annex XV dossier as a whole and the conclusion that octylphenol is to be considered an endocrine disruptor. We would encourage many more of such substances are included in the Candidate in order to ensure a high level of protection of human health and of the environment. However, we have several remarks with regard to the human health side where the conclusion is not satisfactory protective of human health due to the reasons mentioned below;
Taking into consideration the below mentioned new data on endocrine disrupting effects in mammals, the lack data on probable exposure routes and the very high production volume of octylphenol and its occurrence in wide spread applications whereof many close to consumers, the conclusion that should be reached is that octylphenol is indeed an endocrine disruptor also in mammals.
	Thank you for the support

see response to COM 24	
11	2011/10/13	National NGO
Czech Republic	We support the Annex XV dossier as a whole and the conclusion that octylphenol is to be considered an endocrine disruptor. 

Taking into consideration the below mentioned new data on endocrine
disrupting effects in mammals, the lack data on probable exposure routes and the very high production volume of octylphenol and its occurrence in wide spread applications where of many close to consumers, the conclusion that should be reached is that octylphenol is indeed an endocrine disruptor also in mammals.	Thank you for the support

see response to COM 24	
10	2011/10/13	Member State
Sweden	The Swedish CA agrees that 4-tert-octylphenol should be added to the candidate list as a SVHC according to article 57(f) in REACH. We think that the information submitted by Germany clearly show the endocrine disrupting property of 4-tert-octylphenol and that this property should be considered as being of “equal level of concern”. 
In principle we think that for identification of a substance as a SVHC according to article 57(f) due to its endocrine disrupting properties, it is normally irrelevant whether the concern arises from environmental or health data. Therefore we think that it is important that the conclusion in the report is modified to express this, e.g second paragraph (bold text) of the summary. 
We consider that since the endocrine mode of action is via estrogenicity, a hormonal regulatory system relevant for a wide variety of species including humans, there is a general concern for this substance. It is therefore important that both health and environmental aspects are considered in possible future applications for, and decisions on, authorization of the substance.
Furthermore, as there are no studies presented with exposure via inhalation we do not think that s.c and i.p exposure routes can be completely dismissed as irrelevant. The s.c. and to some extent the i.p. routes can serve as a proxy for exposure through inhalation, an exposure route that according to Part II of the dossier is relevant for human exposure.
	Thank you for the support

The conclusion for 4-tert-octylphenol of being a substance of very high concern was drawn on the basis of environmental data. As neither the legal text nor the REACH guidance on SVHC identification states that this conclusion has to be drawn for the human health part as well to place a substance on the candidate list it is considered in accordance with the REACH regulation to propose 4-tert-octylphenol for the candidate list solely based on the conclusion for the environment. 
We disagree with the view that studies with s.c. and i.p. were dismissed in the assessment. All available data on 4-tert-OP were included in the assessment. As no studies with administration of 4-tert-OP by the dermal route or by inhalation were available the adverse affects caused by treatment with 4-tert-OP were identified in studies with oral application. Studies with non-physiological application routes which bypass barrier functions and metabolism at the site of application were used to evaluate the endocrine activity of 4-tert-OP.	
9	2011/10/12	Member State
Belgium	Belgium supports the proposal to identify 4-(1,1,3,3-tetramethylbutyl)phenol, (4-tert-Octylphenol) as SHVC according to article 57 (f) because of its environmental endocrine disrupting properties and subsequent serious effects to the environment (main evidence for fish).
Effects reported do occur at environmentally relevant concentrations in the low μg/l range. Several endpoints, e.g. fecundity, may have significant long-term effects on reproductive success of populations and are shown to be endocrine-mediated. The endocrine mediated effects are highly relevant for a diversity of populations and taxonomic groups, which might irreversibly be affected.	Thank you for the support
	
7	2011/10/12	Member State
Norway	We support the conclusion that OP should be considered as a substance of very high concern because of its ED properties according to REACH article 57 (f), giving rise to an equivalent level of concern as exerted by CMR or PBT/vPvB substances.	Thank you for the support
	
			See also:
Comment 7 attachment (Norwegian comments on OP SVHC dossier.doc).doc		
5	2011/10/12	National NGO
Bund für Umwelt und Naturschutz Deutschland (BUND) e.V. - Friends of the Earth Germany
Germany	In general, BUND supports the Annex XV dossier as a whole and the conclusion that octylphenol is to be considered an endocrine disruptor but we have several remarks with regard to the human health side where the conclusion is not satisfactory protective of human health due to the reasons mentioned below;

Taking into consideration the below mentioned new data on endocrine disrupting effects in mammals, the lack data on probable exposure routes and the very high production volume of octylphenol and its occurrence in wide spread applications whereof many close to consumers, the conclusion that should be reached is that octylphenol is indeed an endocrine disruptor also in mammals.	Thank you for the support

see response to COM 24	
			See also:
Comment 5 attachment (Annex to BUND comments on octylphenole dossier.doc).doc		
4	2011/10/12	International NGO
Center for International Environmental Law (CIEL)
United States	Our organization supports the inclusion of octylphenol (4-(1,1,3,3-tetramethylbutyl)phenol, (4-tert-Octylphenol)) to the REACH Candidate List. 	Thank you for the support	
3	2011/10/12	International NGO
WECF - Women in Europe for a Common Future
Germany	In short we support the Annex XV dossier as a whole and the conclusion that octylphenol is to be considered an endocrine disruptor but we have several remarks with regard to the human health side where the conclusion is not satisfactory protective of human health due to the reasons mentioned below;
Taking into consideration the below mentioned new data on endocrine disrupting effects in mammals, the lack data on probable exposure routes and the very high production volume of octylphenol and its occurrence in wide spread applications whereof many close to consumers, the conclusion that should be reached is that octylphenol is indeed an endocrine disruptor also in mammals.	Thank you for the support

see response to COM 24	
2	2011/10/12	Member State
Denmark	We agree that 4-tert-octylphenol is an endocrine disruptor and should be included on the candidate list as a SVHC of an equivalent level of concern (article 57f). 

However, we do not agree that the substance is only an endocrine disruptor of concern for the environment, and we believe that the conclusion should be amended. 

As proposed in the Danish "Establishment of Criteria for Endocrine Disruptors and Options for Regulation" of 17 May 2011, we would assess the endocrine properties of a chemical in a weight of evidence approach including data from both ecotoxicity studies and from studies of relevance for human health (mainly in rodents). Only if there is clear evidence that the endocrine disruption is not of relevance for human health, we would conclude that the effects are only relevant for the environment. This is in our view not the case with 4-tert-octylphenol, which induces endocrine disruptive effects in a variety of species and test systems (e.g. in in vitro studies, in fish, in amphibians and in rodents) and which seem to act as an endocrine disruptor mainly through the estrogen receptor, which has been well conserved during evolution and can be found in a wide variety of species, including humans.

Considerations regarding potency, whether the substance is or should be classified as a reproductive toxicant or a carcinogen and whether the endocrine effect is the lead effect is in our view not relevant in the clarification of whether the substance is an endocrine disruptor (page 153-155), but should be included in subsequent analysis of risk management options. Effects induced through subcutaneous or interperitoneal routes of exposure should not be disregarded, but should be included in the overall weight of evidence, considering that these routes of exposure (which bypass metabolism in the small intestine and the liver) could be indicative for effects induced through dermal exposure or inhalation.

In conclusion, we believe that both environmental and human health concerns should be reflected in the conclusion regardless of whether the evidence originates from ecotoxicity or toxicity studies. This is of utmost importance, since it might impact the further considerations regarding possible Authorisations as well as the future possibility to regulate combined exposure to endocrine disrupters.
	Thank you for the support

The conclusion for 4-tert-octylphenol of being a substance of very high concern was drawn on the basis of environmental data. As neither the legal text nor the REACH guidance on SVHC identification states that this conclusion has to be drawn for the human health part as well to place a substance on the candidate list it is considered in accordance with the REACH regulation to propose 4-tert-octylphenol for the candidate list solely based on the conclusion for the environment.
The reference to the DK criteria is acknowledged but it is not the intention of the dossier to anticipate the outcome on the criteria discussion for the assessment of substances that cause adverse effects mediated by endocrine mechanisms. Therefore, we would like to draw the attention to the particular case, i.e. the proposal to add 4-tert-OP to the candidate list.	
1	2011/10/10	Trade union
European Trade Union Confederation
Belgium	ETUC supports the identification of 4-(1,1,3,3-tetramethylbutyl)phenol, (4-tert-Octylphenol) as SVHC.	Thank you for the support	

Specific comments on the justification
Number	Date	Submitted by (name, Organisation/
MSCA)	Comment	Response	
27	2011/10/13	International NGO
Health and Environment Alliance
Belgium	Whilst we understand that the WHO IPCS definition of an endocrine disrupting chemical is important to take note of, we nevertheless disagree that this exact definition should be used in REACH.  The WHO  IPCS definition is not expressly cited in the REACH text, and we feel the specification 'CONSEQUENTLY' is too high a level of proof to cohere with the REACH phrase for equivalent concern chemicals of 'PROBABLE' serious effects.  We also find it strange that given the adverse effects found in animals, that the dossier does not conclude that Octylphenol is of equivalent concern for its endocrine disrupting properties to human health. 	Thank you for your comment with regard to criteria for identifying substances as SVHC based on the endocrine disrupting properties. They are appreciated and we are sure that they will provide valuable input in the further discussion on criteria. Please note that the dossier does not aim at pre-empting this discussion.

See also response to COM 24 and to COM 10 in General comments.	
26	2011/10/13	Other contributor
R.I.S.K. Consultancy
Belgium	Please see the .pdf version of my comments, attached.	Thank you for your support and your comments.

For the assessment of 4-tert-octylphenol an extensive literature search was carried out. As can be found in the report, publicly available literature as well as unpublished study reports - as far as available - were included in the assessment. The attached references are known to the dossier submitter, but were not included due to unclear substance identity in the study or different octylphenol isomer studied. 

With regard to the three ecotoxicological studies mentioned in your comment it turned out that they were already included in the dossier (results by Croteau et al, (2008) are cited in Croteau et al (2009).	
			See also:
Comment 26 attachment (OP_EDC_SVHC-tt.pdf).pdf		
24	2011/10/13	National NGO
CHEM Trust
United Kingdom	Environment – Chapter 6.2.1
The justification for this chemical having endocrine relating properties that give rise to probable serious effects on the environment and an equivalent level of concern as other Article 57 chemicals, such as CMRs and PB(T)s is well presented.  We consider that it is particularly the likelihood of delayed irreversible effects due to endocrine disrupting chemicals de-railing normal development during sensitive windows of exposure that gives rise to equivalent levels of concern as the PB(T) chemicals.  This is because effects may not become evident until the animal reaches maturity, such that, like the vPvB chemicals, if adverse effects do come to light in future they will be ongoing for many years.

We understand the approach taken in the dossier with regard to leaving open some of the issues relating to the criteria, but it needs to be made very clear that potency should not play a part in the identification of a chemical with ED properties.  It is only the intrinsic properties of ED that must be fulfilled. Consideration of the potency is only possible at a later stage when the risk management is considered.  
We consider that this chemical is a potent ED in wildlife, however, this should not set a precedent that such a high degree of potency is required for subsequent chemicals, because as noted above, potency should not be considered at this stage.  Furthermore, discussions on lead effect are also not appropriate.  (For more detailed argumentation, please see the enclosed document from CHEM Trust which had significant input from WWF-European Policy Office “CHEM Trust’s Contribution to the Ongoing Debate on Criteria for EDCs – September 2011”

Page 143:
CHEM Trust does not agree that “a substance should fulfill at least the definition of an ED provided by IPCS/WHO in order to be considered as of equivalent concern based on ED properties. The legal wording is “having endocrine disrupting properties.” The problem of the IPCS/WHO definition is that it requires considerable data to prove the exact mechanism behind an adverse effects. It can take years to establish this and while this is rather clear for octylphenol which has been well investigated, this should not be the level of proof required for all future EDs.  Moreover, the IPCS definition requires ‘consequently causes adverse effects’ whereas the legal text of REACH requires ‘probable serious effects’.  Here it can be seen that the word ‘probable’ allows some degree of uncertainty.  It is imperative that the requirements of the legal text are used rather than some other definition.

(Again, see also the uploaded position paper on EDC criteria written by CHEM Trust with input from WWF European Policy Office “CHEM Trust’s Contribution to the Ongoing Debate on Criteria for EDCs – September 2011”)

CHEM Trust considers that the data is well presented to show OP has ED properties in many species, including fish, amphibians and invertebrates.

Human health – Chapter 6.2.2
CHEM Trust considers that it would be better to re-structure this chapter and not mix the discussion on potency and lead effect with the identification of the ED properties. There are normally no potency considerations in the identification of CMRs, where instead it is based on the level of evidence, so it is inappropriate to include potency considerations in the identification of ED properties.  Furthermore, neither the IPCS/WHO definition, nor the OECD draft Guidance document requires any consideration of the potency of the chemical for it to be determined to have ED properties.

CHEM Trust considers that the data presented provide clear evidence that octylphenol does exhibit ED properties in vitro and in vivo.  As the prepubertal oral uterotrophic study (Laws et al., 2000) showed an increase in uterine weight this is a clear indication of ED properties in an intact animal, which is of relevance for adverse effects in young girls.

The LOEL was lower using the s/c route of exposure which indicates that when there are exposure routes that bypass metabolism there should be added concern for estrogenic effects.  We consider that information gained using the sub-cutaneous (s/c) route of exposure should inform the overall assessment.

CHEM Trust considers that given the conservation of the endocrine system in the evolution of vertebrate animals, the default should be to assume that a chemical with ED properties in one vertebrate species will also have such properties in others.  If data contradict this, then the possible reasons should be examined.  For example, in fish it can be seen that OP is clearly active as an ED.  However, aqueous exposure of fish is via uptake through the gills and skin, which essentially bypasses metabolism in the small intestine and liver, and allows direct entry to the blood stream.  Therefore, it is possible to visualize a scenario in which the endocrine activity of a substance can be markedly decreased by metabolism after oral exposure.  This could lead to no or little endocrine related findings in the oral studies in mammalian animals, whereas endocrine activity may manifest itself in fish where the substance is not metabolized before reaching the site of action or the relevant organ.   If this were the case with OP, then the oral exposure of mammals would not be representative of the inhalation or dermal routes of exposure, which also bypass metabolism.  There are no data for either exposure routes for this chemical, and without such data – it should certainly not be concluded that this is not a potent ED chemical in humans.   It is noted that this chemical is found in dust and in biomonitoring of housekeepers, suggesting that inhalation is a route of concern.  Moreover, it is also found in textile workers suggesting that either or both dermal and inhalation exposure may be important.  We note that OP is found in many imported textiles, such that dermal exposure should be a concern.

With regard to a related chemical, there was a discussion many years ago under the Existing Substances Regulation, where there was concern for a potential risk with regard to the use of nonyl phenol ethoxylate, in that it was suggested that the dermal application of a de-greasing hand cleaner might result in significant exposure to nonyl phenol, a related compound.  This highlights the need to consider the likely effects from dermal exposure and the precedent for this.

An alternative explanation for the low potency for ED effects seen in oral studies might be either that too high a dose had been used, or that the tests used were not sensitive enough.  CHEM Trust suggests that when OECD study results are discussed, then the year that the study was conducted should always be noted.  This is because, for example, it might be that a chemical had been tested using the 2-Gen (TG 416) which was in use prior to the last update in 2001, when it was made more sensitive to chemicals with ED properties.  It should also be noted that even this 2001 updated version of the TG416 is still missing many important end points which would make it more sensitive for ED.

We consider that the most likely explanation for the results which indicate lower potency for ED in oral studies in rodents is that there is some metabolism in the gut. CHEM Trust therefore considers that the activity in mammalian animals using the sub-cutaneous route should indeed be taken into account, particularly as it is found in humans in some biomonitoring studies, and dermal and inhalation exposure via dust is likely.  Nevertheless, we stress that potency considerations are not relevant for the identification of ED properties.

It should also be recognized that for potency considerations, test methods for ED effects need to be improved.  For example, given the effects of estrogenic compounds on the mammary gland, we consider that tests need to be further improved with regard to including looking for effects of chemicals with ED properties on the mammary gland of both females and males.  This is because in some chemicals tested to date, mammary gland development in males and females is altered at lower doses than the levels that cause changes in other tissues. The effects that have been observed after disrupted mammary gland development include impaired lactation and increased susceptibility to cancer, both of which are of potentially great public health significance. Key gaps in testing include lack of early life exposure in tests for carcinogenic effects; lack of assessment of lactation function other than nonspecific measures; and inadequate examination of mammary gland morphology and pathology.

Therefore overall, we consider that there are sufficient data for OP to be considered to have ED properties relevant for both the environment and human health.  	Thank you for your comment with regard to criteria for identifying substances as SVHC based on the endocrine disrupting properties. They are appreciated and we are sure that they will provide valuable input in the further discussion on criteria. Please note that the dossier does not aim at pre-empting this discussion.

The IPCS definition is an internationally elaborated definition of endocrine disruptors, in which ‘Endocrine disruption’ is agreed to be a mechanism/mode of action and not an adverse effect per se. The German CA is of the opinion that this definition should be used under REACH to warrant an equal treatment of ED substances not only throughout the various legislations in the EU but also internationally. 

See also response to COM 24 and to COM 10 in General comments.

	
			See also:
Comment 24 attachment (CHEM Trust Position on EDC Criteria - Sept11.pdf).pdf		
23	2011/10/13	International NGO
WWF European Policy Office
Belgium	Chapter 6.2.1 Environment (pages 143 – 149)
All in all this part is well described and convincing evidence has been presented on the endocrine disrupting properties as well as on the scientific evidence of probable serious effects to the environment which give rise to an equivalent level of concern.

However, WWF does not agree that “a substance should fulfill at least the definition of an ED provided by IPCS/WHO in order to be considered as of equivalent concern based on ED properties.” (page 143). The REACH text in article 57f speaks of substances “having endocrine disrupting properties.” The IPCS/WHO definition requires considerable amount of data to prove the exact mechanism of action leading to an adverse effect. It can take years to establish this knowledge and while it is known for octylphenol which has been well investigated, this should not become the standard expectation for all future identification of chemicals with endocrine disrupting properties.  In our view it should also be made clear that the considerations on potency and lead effect should not play a role in the identification of ED properties (see also comments relating to the chapter 6.2.2 on human health). 

For more details, please see also the uploaded position paper on EDC criteria written by CHEM Trust with input from WWF European Policy Office.

Chapter 6.2.2 Human health (pages 150-155)

It is proposed to structure the chapter in the same way as 6.2.1. This would have the advantage of separating the description of the ED properties from the justification of demonstrating scientific evidence of adverse effects similar to CMR and PBT. 

In our view considerations on potency and lead effect should not play a role in the identification of the ED properties. There are normally no potency considerations in the identification of CMRs, where it is instead done based on the level of evidence, and therefore it is inappropriate to use them in the ED identification.

The chapter presents clear evidence that 4-tert-octylphenol does exhibit ED properties in vitro and in vivo (demonstrated in many species). We doubt that it is justified to conclude that this is NOT relevant for human health. The fact that the endocrine system has been conserved throughout the evolution of vertebrates suggests that  - on the contrary - it must be regarded as relevant.

For more details, please see also the uploaded position paper on EDC criteria written by CHEM Trust with input from WWF European Policy Office.	Chapter 6.2.1 Thank you for your support. 

Thank you for your comment with regard to criteria for identifying substances as SVHC based on the endocrine disrupting properties. They are appreciated and we are sure that they will provide valuable input in the further discussion on criteria. Please note that the dossier does not aim at pre-empting this discussion.

See also response to COM 24 and to COM 10 in General comments.
	
			See also:
Comment 23 attachment (CHEM Trust Position on EDC Criteria - Sept11.pdf).pdf		
21	2011/10/13	Member State
Ireland	4-tert-octylphenol is proposed to be identified as a substance of equivalent concern according to Article 57 (f) because of its environmental endocrine disrupting properties and subsequent serious effects to the environment. The Irish CA agrees that the substance meets the criteria for identification as an SVHC in accordance with Article 57 (f) of REACH.	Thank you for your support	
20	2011/10/13	Industry or trade association
European Council for Alkylphenols and Derivatives (CEPAD)
Belgium	2.0	The properties of OP fall well short of the Article 57 criteria for CMR Cat. 1A or 1B, PBT, vPvB.  

Relates to Sections 4, 5 and 6 of the Annex XV Report

2.1	The Annex XV Report concludes that OP does not meet the criteria for probable serious effects for SVHCs as defined under CMR Cat. 1A or 1B; it also does not meet criteria for the weakest category, CMR Cat. 2.

Criteria to determine whether a substance should be identified as a “CMR Cat. 1A or 1B” or Cat. 2 are given in the EC Regulation No 1272/2008 (EC, 2008, Dec. 16) on classification, labeling and packaging of substances and mixtures.  Under this regulation, “reproductive toxicity includes adverse effects on sexual function and fertility in adult males and females, as well as developmental toxicity in the offspring”. The subcategories for reproductive toxicants and their respective criteria are defined under EC Regulation No 1272/2008 as: 

Category 1A (Known Human Reproductive Toxicant), which is largely based 
         on evidence from humans.  

Category 1B (Presumed Human Reproductive Toxicant), which is largely based 
                      on data from animal studies.  

EC Regulation No 1272/2008 requires that “the data shall provide clear evidence of an adverse effect on sexual function and fertility or on development in the absence of other toxic effects, or if occurring together with other toxic effects the adverse effect on reproduction is considered not to be a secondary non-specific consequence of other toxic effects. However, when there is mechanistic information that raises doubt about the relevance of the effect for humans, classification in Category 2 may be more appropriate.

Category 2 (Suspected Human Reproductive Toxicant), which is intended  for substances where there is some evidence from humans or experimental animals, possibly supplemented with other information, of an adverse effect on sexual function and fertility, or on development, and where the evidence is not sufficiently convincing to place the substance in Category 1. 

If deficiencies in the study make the quality of evidence less convincing, Category 2 could be the more appropriate classification. Such effects shall have been observed in the absence of other toxic effects, or if occurring together with other toxic effects the adverse effect on reproduction is considered not to be a secondary non-specific consequence of the other toxic effects.

Substances that are CMR Cat. 1A and 1B are of concern because there is sufficient data to determine that they are “known” or “presumed to be” carcinogenic, mutagenic or toxic to reproduction. Carcinogens induce or increase the incidence of cancer. Mutagens cause a permanent change in the amount or structure of genetic material in cells.  Reproductive toxicants cause adverse effects on sexual function and fertility or on development off offspring in the absence of other toxic effects. 

2.2	  OP is not a PBT or a vPvB

A PBT or vPvB substance is one that meets the clear definitions according to Annex XIII of Regulation (EC) No 1907/2006 and to the guidance on information requirements and chemical safety assessment Chapter R.11 (PBT Assessment, ECHA (2008)).  The European Center for Ecotoxicology and Toxicology of Chemicals (ECETOC) Technical Report on the Risk Assessment of PBT Chemicals states: 

 “chemicals categorized as meeting the P or vP criteria are of particular concern because of their predicted long environmental half-lives. As a consequence, successive releases over time could result in a build-up in the environment if the rate of release is higher than the rate of disappearance. Additionally, when distribution within the environment occurs, the presence of such chemicals could be observed on a wider geographical scale, therefore spreading the possible concern to remote areas. Moreover, when a toxic substance is not only persistent in the environment, but also able to bioaccumulate, the potential for exposure to living organisms in a higher trophic level to a particular substance is increased. This may lead to possible adverse sublethal effects from chronic exposure that would not have been predicted from standardised laboratory tests (ECETOC, 2006)." 

While there are many substances that may be persistent, bioaccumulative, or toxic, it is the unique combination of all three properties that defines the overall hazard potential of a PBT. The threat posed by a PBT substance is that it enters the environment, does not degrade, cannot be metabolized by the organisms that ingest it, and poses toxicity to those organisms. A substance designated “very persistent and very bioaccumulative, or vPvB, have degradation properties that are even slower than for meeting the persistence criteria for a PBT substance. In addition, vPvB substances have even greater bioaccumulation criteria than for a PBT substance and  are considered to be so persistent and bioaccumulative, more so than a PBT substance, that they are treated in the same regulatory manner as CMR and PBT substances, regardless of any toxicity concerns. 

2.2.1	Persistence Assessment (P)

While the Annex XV report includes some discussion of degradation data for OP in these media, a more complete assessment of the expected environmental persistence in alignment with the REACH submission for OP (IUCLID, 2010) is given below. 

According to Annex XIII, a substance does fulfill the criteria “persistent (P)”, if: 
-      the half-life in marine water is higher than 60 days, or 
-      the half-life in fresh- or estuarine water is higher than 40 days, or 
-      the half-life in marine sediment is higher than 180 days, or 
-      the half-life in fresh- or estuarine water sediment is higher than 120 days, or 
-      the half-life in soil is higher than 120 days. 

Simulation tests for the biodegradation of OP and degradation intermediates are available for all environmental compartments listed above except for soil, which can be addressed using read-across from the analogue substance nonylphenol (NP).  Half-lives of OP biodegradation in the different environmental compartments are summarized in Table 1. The available data demonstrate OP is biodegradable in river bed sediments, marine sediments, surface waters, and soil under aerobic conditions. Furthermore the data support a conclusion that OP does not meet the criteria for persistency in any of these environmental compartments. 

While OP was found to not significantly biodegrade under anaerobic conditions, many organic compounds degrade slower under low temperature, nutrient-poor or oxygen-limited conditions (see for example Boethling and Mackay, 2000). The data for OP are consistent with the findings for other such substances.  OP does degrade in the environmental compartments in which organisms reside. These compartments include freshwater and marine water and sediment and soil. Because OP does degrade in these compartments, it is not expected to build up in the environment and thus does not pose the concern caused by truly long lasting and persistent substances. Therefore, OP is not considered “persistent” or “very persistent” according to Annex XIII criteria.

See Table 1. Summary of Aerobic Biodegradation Data for Octylphenol (Provided in attached comments document) 

2.2.2 	Bioaccumulation Assessment 
The Annex XV Report presents a useful summary of the available data on the bioaccumulation potential of OP. The report gives a range of experimentally determined bioconcentration factors (BCF) of 46 to 471. 
According to Annex XIII, a substance does not fulfill the criteria of “bioaccumulative (B)” or “very bioaccumulative (vB)” if the BCF are below 2000 or 5000, respectively.  The EU Commission reviewed OP in the course of “Interim Strategy for Management of PBT and vPvB Substances” and confirmed that this compound does not fulfill PBT/vPvB criteria (ECB, 2006).  In the Annex XV report for OP, the stated conclusion is that the bioaccumulation potential of OP is low to moderate. 
Overall, the weight of evidence for OP metabolism suggests a low potential for bioaccumulation due to rapid elimination via liver hydroxylation, glucuronidation, and sulfation. Bioaccumulation is only observed if these detoxification pathways are saturated by excessive doses  (IUCLID, 2010) 
The concern posed by bioaccumulative substances that meet the PBT criteria is that they may have the potential to also biomagnify (build up in food-chains and cause increasing exposure with higher trophic levels, including humans, aquatic mammals, and birds). As the studies cited above show, and as noted in the ECB (2006) review, mammals and fish are capable of rapidly metabolizing OP; therefore this compound has low to moderate bioaccumulation potential, which eliminates the concern of increasing accumulation within food webs that are relevant to compounds that meet the PBT or vPvB criteria.  
In summary, OP is neither a “bioaccumulative” or “very bioaccumulative” substance according to Annex XIII criteria.

3.0	The Report does not adequately support its argument that OP is a potent endocrine disruptor. 
Relates to Sections 4, 5 and 6 of the Annex Report and to the Report in genera

3.1 	OP is considerably less potent than the natural estrogen, 17-estradiol (E2) and synthetic (EE2) estrogens based on in vitro estrogen activity screening tests.

Valuable information about the relative potency of endocrine active compounds relative to the known potent estrogens E2 and EE2 are provided by various in vitro screening tests.  As summarized in the table below (Table 2), relative potencies of either E2 or EE2 and OP ranged from 0.000032 to 0.0024 (400 to 31,000 fold less potent) for vitellogenin (VTG) induction and estrogen receptor binding in hepatocytes from several species of fish. As an example, using an in vitro assay with rainbow trout hepatocytes, Jobling and Sumpter (1993) reported a relative potency of OP to E2 of 0.000037, or 27,000 times less potent than E2

3.2	OP is also considerably less potent than E2 and EE2 based on in vivo studies that examined endocrine-sensitive apical endpoints.  

The Annex XV Report greatly overstates the estrogenic potency of OP and goes to great lengths to calculate effect ratios for various endpoints from studies with OP and E2 or EE2 in an attempt to assess its potency relative to these highly estrogenic compounds.  It should be noted that apical endpoints reflect the integration of multiple different modes of action at the biochemical level; therefore calculations of the relative estrogenic potency of compounds based on No Observed Effect Concentrations (NOEC) for apical endpoints does not necessarily provide a specific measure of estrogenic potency.   In addition to this general issue, the following comments raise other concerns about the approach taken in this section of the Report. 

Some of the NOEC ratios presented in the Annex XV Report are not at all useful because they were determined from studies that used only one or two concentrations of either the hormone or OP. In other words, the Report compares NOEC for an endpoint in studies where there were no effects in the study (i.e., no reported LOEC) to a NOEC that did have an LOEC.  This approach is not scientifically founded and provides no useful information about the estrogenic potency of OP relative to E2 or EE2.  For OP, roughly half of all NOEC reported in the Report were actually “no effects at the highest concentration tested”; therefore the actual NOEC could be much higher.  It is inappropriate to represent such results as meaningful for E2 or EE2 to OP comparisons. 

If there is interest in comparing the relative potency of two compounds based on NOEC data for apical endpoints, the most meaningful approach is to compare NOEC from studies where an LOEC was found based on dose-response.  The best examples of this are from the life cycle studies with EE2 and OP and zebrafish conducted by Wenzel et al. (2001). The tests with EE2 and OP were done nearly identically. Numerous identical endpoints were recorded.  NOEC for each endpoint that also recorded an LOEC for both EE2 and OP were F0 juvenile length, F0 time to spawn, F0 eggs per female per day, and F0 fertilization success. The NOEC ratios for OP:EE2 are 0.000025 (or 40,000-fold less potent) for each endpoint. If the ratios of the LOEC for the same endpoints are used, the ratios are 0.000031 (or 32,000-fold less potent). Thus EE2 is 32,000 to 40,000 times more potent than OP, based on the most important endpoints from the full life cycle study. 
  
Relevant studies of shorter term duration confirm these relative potency results (see summary Table 2 below). Using NOEC that are consistent and treatment related, findings from the study with guppies by Toft and Baatrup (2003) showed that estrogenic hormones were 2,000-fold more potent than OP in regards to sex ratio and 1,000-fold more potent than OP in regard to length and colorization. Van den Belt et al. (2001) showed that estrogenic hormones were 20,000-fold more potent than OP for percent spawning and 2500-fold more potent than OP for ovarian-somatic index (OSI). 

3.3	Relative potencies for OP:E2 or OP:EE2 are similar with in vitro and in vivo testing

Relative potencies are shown in the summary table (Table 2) for in vitro and in vivo tests using studies on E2 or EE2 and OP for which direct comparisons of effects could be made. There is little difference between the range of in vitro (400 to 32,000-fold) and in vivo relative potencies (1000 to 20,000-fold). The ranges of relative potencies of apical (fertilization success, spawning success, sex ratio, body length) and secondary endpoints (coloration, OSI, vitellogenin (VTG) induction) are also similar (1000 to 40,000-fold) versus 1000 to 15,000-fold). Among apical endpoints, the single lowest relative potency was 1000-fold for body length and the highest was 37,000 for fertilization success in a life cycle test with zebrafish. 

Overall the in vitro and in vivo data show that OP is considerably less potent than either E2 or EE2. These data show that the statement in the Report at Section 6.2.1.2 that OP is nearly as potent as E2 (within a factor of 2 to 28-fold) is clearly incorrect and an artifact of using unreliable studies and improper analysis of the data.   

See Table 2. Summary of Relative Potency Ratios for E2/OP and EE2/OP in attached Comments 

4.0	Examination of the estrogenic potency and aquatic toxicity of natural estrogen (E2) and synthetic estrogen (EE2) provides an understanding of what constitutes a highly potent estrogen.  

Relates to Sections 4, 5 and 6 of the Annex XV Report 

The Annex XV Report emphasizes the estrogenic activity of OP as the basis for “equivalent concern” in its assessment of the compound’s aquatic toxicity. In light of this concern, it is of use to first examine effects on aquatic organisms from known endocrine active substances for which the mode of action is indisputable – natural estrogen (E2) and a synthetic and therapeutic estrogen (EE2). Relevant studies are summarized in the table below (Table 3) and more fully presented with experimental details in the attached Table A.  E2 and EE2 have been found to be of generally similar potencies to each other. EE2/E2 ratios range from 0.19 to 1.9 depending on the screen used (Kidd et al., 2007).  E2 and EE2 are especially potent when compared to the weak estrogenic activity of OP. 

The natural and synthetic estrogens E2 and EE2, respectively, cause significant and severe effects on freshwater and saltwater fish. As described in more detail below, a signature set of aquatic effects from E2 and EE2 may include:  complete feminization of male gonads; induced failure to differentiate into males; reduced reproduction capabilities; severe histological effects on gonadal tissues; and even collapse of whole populations in a lake continuously dosed with EE2 for three years. 

A series of short term reproduction assays showed that survival, reproduction, fertilization, spawning and hatch success, and sex ratio can be significantly altered at concentrations down to 0.005 μg/L. Secondary endpoints such as gonadal weights and histopathology, testis-ova, and kidney lesions were affected at even lower concentrations, down to 0.0002 μg/L.

Life cycle studies with fathead minnow and zebrafish that included exposure to EE2 over the full life cycle showed similar effects at 0.001 to 0.016 μg /L and 0.0003 to 0.010 μg /L, respectively. Second generation NOEC were seen at concentrations as low as 0.0002 to 0.0003 μg /L (Langer et al., 2001; Wenzel et al., 2001). Secondary endpoints including secondary sex characteristics, testis-ova, gonadal histopathology, and VTG had NOEC in the 0.001 to 0.004 μg /L range. In both studies, and presumably due to the strong estrogenic potencies of these compounds essentially complete feminization of males and/or male characteristics were reported. 

In order to differentiate the relationship of endpoint effects caused by a narcotic mode of action and those mediated by an estrogenic mode of action, Wenzel et al . (2001) examined the ratio of the acute LC50 of EE2 (1,700 μg /L) and the most significant reproduction NOEC (fertilization success, NOEC 0.0003 μg /L). The acute-to-chronic ratio of the two is 5.73 million. This very high ratio is indicative of the mode of action associated with the synthetic estrogen EE2 and very different from the ratios that are calculated with OP (30 to 63) as is discussed below. 

In a whole lake study, Kidd (2007) and colleagues dosed a whole lake (0.005 to 0.006 μg/L) with EE2 for 3 years. Fathead minnow populations became completely feminized, reproduction halted, and the population collapsed. The abundance of pearl dace fish was also decreased. These two species of fish are predominant in the lake. As noted in laboratory studies, severe effects on gonadal tissues were found. Male gonadal tissue development was stunted or halted, edema was noted in ovarian tissues and lesions were found in kidney tissue. 

See Table 3 Summary of Aquatic Toxicity Studies for 17β-ethynylestradiol (EE2) and 17β-estradiol (E2) in attached comments. 
In summary, the effects on aquatic organisms by potent estrogenic substances such as E2 and EE2 collectively add up to a predictable, cohesive signature of effects, which are consistent with the known mechanism of action for these compounds. Natural development of male gonadal tissue is halted, differentiation into male characteristics is blocked, male secondary sex characteristics are prevented from developing, and gonadal tissues are severely compromised and damaged, reproduction within natural populations can be halted, and effects that are mediated by estrogenic activity occur at concentrations that are millions of times lower than those where narcotic effects occur. As a scientifically rigorous review of relevant studies shows (see Section 6.0 below), the effects of OP on aquatic organisms are very different from the effects from potent estrogens.  

5.0	The scientific evidence presented in the Annex XV Report for OP is in some cases  based on invalid studies and flawed study interpretation and the overall assessment does not adequately support a case that this compound rises to a level of concern that is equivalent to a CMR Cat.  1A, 1B, PBT or vPvB compound. 

Relates to Sections 5 and 6 of the Annex Report 

The bulk of the Annex XV Report is devoted to presenting a case that OP is a substance of “equivalent concern”; however the scientific analysis used to make that case is circumstantial, simplistic, incomplete, and in some cases wrong. Specific examples of these problems are given in the discussion of the individual studies below. A full analysis of the available aquatic toxicity data for this compound does not give rise to a conclusion that serious aquatic effects are probable or would be comparable to a Cat. 1A or 1B CMR, PBT or vPvB substance.  

5.1	There are problems with study quality scoring in the Annex XV Report on OP

In Chapter 5 of the Annex XV Report for OP, the aquatic hazard assessment is presented. The assessment introduces a number of studies mostly with fish, amphibians, and invertebrates and identifies potential findings that might be indicative of endocrine activity. Prior to analysis of any given study, the authors of the report gave each study a study quality score according to the principals of Klimisch (1997). The scoring of the various studies by the authors of the report was inappropriate in many cases. Types of errors include scoring studies as valid (scores 1 or 2) that used no statistics, lacked replication, or had poor control performance. Additionally, several studies were scored as “not valid” (3), but the studies were used anyway in the Report. Studies properly scored as “not valid” (3) cannot be used in a hazard assessment. All studies presented in the Report are captured in the attached Table B and have been re-scored following these practices.

5.2	There are problems with statistical significance and interpretation of LOEC. 

A second issue with the analysis of individual studies is the use in the Report of “Starts at”. In other words, presenting results, as positive findings, that were in fact not positive because they were not statistically different from controls. All the OECD guidance for conducting of aquatic studies, from screening studies to definitive multi-generation studies, call for adequate replication of controls and treatments such that meaningful statistical analysis can be performed. Effects are determined for any endpoint only by demonstrating with proper statistical analysis that controls and treatments are different. In addition, findings of statistical significance must be consistent and dose-related in order to establish NOEC and LOEC. For instance, statistically significant results for an endpoint at a middle dose, but not at higher or lower doses is not considered a consistent and dose or treatment related finding. The intention of the experimental designs of the OECD aquatic testing guidance when toxicity is expected is to test a series of concentrations in order to obtain a dose-response curve. The treatments are each compared using proper statistical analysis to controls and NOEC and LOEC obtained if possible. 

All studies presented in the Report are captured in the attached Table B and have been re-evaluated following these practices. The re-evaluation of individual studies includes the determination of all statistically significant endpoints that are consistent and treatment related. 

6.0	A scientifically rigorous evaluation of the “valid” or “use with care” studies on the aquatic toxicity of OP shows a consistent range of effect concentrations across species and taxonomic groups as well as across endpoints and sensitive life stages.  

Relates to Sections 5 and 6 of the Annex Report 

	6.1 	Fish

Aquatic toxicity results with OP are available for several species of fish (see the summary table below (Table 4) and attached Table B for fuller descriptions of the studies). For the screening or short term tests, specific experimental conditions varied, but adults were exposed generally from 21 to 60 days. In other tests, fertilized eggs were used to initiate the test, ensuring exposure at early and sensitive portions of the life cycle. During or following exposures numerous apical and secondary endpoints were measured. Apical endpoints included survival, growth and development, and reproduction endpoints. Many of the studies conducted reproduction trials either during or following exposure. Reproduction endpoints included egg production, spawning success, fertilization success, and hatching success. Growth and development endpoints included embryo-larval and juvenile growth, gonopodium length, and sex ratio. Secondary endpoints included those at the biochemical, plasma, or tissue level of organization. These include concentrations of vitellogenin (VTG), gonadal somatic indices, testis-ova, gonadal histopathology, and histology of other organs. 

Screening and Short Term Reproduction Studies – For these types of studies, across all apical endpoints, NOEC ranged from 10 to 1000 μg/L, while secondary endpoint NOEC ranged from 1 to 200 μg/L. By category of apical endpoint, reproduction and growth and development endpoints had similar range of NOEC (12 to 1000 and 12 to 900 μg/L, respectively), while survival NOEC ranged from 10 to 300 μg/L. Survival and secondary endpoint NOEC were similar at 10 to 300 and 1 to 200 μg/L, respectively. 

Life Cycle Tests – Life cycle tests represent the most thorough means of evaluating exposure to fish at all critical windows of their life cycle. Fully valid life cycle tests have been performed with Japanese medaka and zebrafish. 

The life cycle study with Japanese medaka was conducted by the Japanese Ministry of the Environment (Japan MOE 2002) and was not available for review. The results discussed here were presented in the Annex XV report. This study exposed newly fertilized eggs to OP throughout development to adults, followed by reproduction and early life stage exposure. The study analyzed the apical endpoints of F0 adult survival, fecundity, fertilization, hatch success, time to hatch, F0 adult growth, and F1 juvenile growth. In addition, the secondary endpoints of testis-ova and VTG induction were measured. No effects on survival, hatch success, time to hatch or F0 and F1 growth were found (highest concentration tested of 82.3 μg/L). Fecundity and fertilization were reduced at 82.3 μg/L, but not at 30.4 μg/L. Thus, the overall lowest NOEC for the study based on apical endpoints was 30.4 μg/L OP. NOEC for the secondary endpoints of testis-ova and VTG induction were ≤9.9 μg/L. Secondary endpoints such as biomarkers are generally more sensitive than apical-type endpoints, due in part to compensatory mechanisms that exist at increasing levels of biological organization (Toft and Baatrup, 2001). This was shown, for instance, in a partial life-cycle reproduction study with sheepshead minnow with EE2 (Zillioux et al., 2001) in which effects on the secondary endpoints testis-fibrosis and testis-ova were reported to occur at 2 to 20 μg/L, while effects on the apical endpoints of reproductive success and hatching success occurred only at 200 μg/L. 

The second life cycle test with zebrafish exposed newly fertilized embryos to OP through development to adult, followed by reproduction and early life stage testing. Apical endpoints of F0 and F1 early life stage survival and growth, F0 sex ratio, F0 time to spawn, F0 egg production, and F0 fertilization were measured. No effects at any concentration up to the highest tested of 35 μg/L were found on F0 embryo-larval survival, F0 larval length, F0 juvenile survival, F1 embryo-larval survival, or F1 embryo-larval length.  Reduced F0 juvenile length, F0 time to spawn, F0 egg production, F0 fertilization, and F0 sex ratio (fewer males) were reduced at 35 μg/L, but not at 12 μg/L. No secondary endpoints were recorded. These data represent scientific evidence that OP does not cause “probable serious effects” to the environment that are equivalent to CMR Cat. 1A and 1B, PBT, or vPvB substances, because the concentrations of OP required to cause adverse effects with population-level relevance in these fish species are several orders of magnitude above the concentrations of OP that are commonly measured in the environment (discussed further below).

See Table 4. Summary of Aquatic Toxicity Studies with Fish for Octylphenol (OP) in attached comments 

6.2   Amphibians

There have been several studies with amphibians and OP (summarized in attached Table C) that have been scored for reliability as (1) “valid” or (2) “use with care”. Studies scored as either (3) “not valid” or (4) “insufficient information” should not be used for hazard assessment. Explanation for reliability scoring of studies cited in the Report is given in attached Table C. 

Bevan et al. (2003) exposed embryos of African clawed frogs to OP. Effects on body length were noted at 100, but not 20 μg/L, while developmental abnormalities were reported at 1000, but not 100 μg/L. The study with the same species by Kloas et al. (1999) is considered not valid as the authors failed to control temperature during the test, had no analytical confirmation of test concentrations, had skewed sex ratio in controls, and used improper statistics. The study with the same species by Van Wyk et al. (2003) is considered not relevant as the authors employed intraperitoneal injection of OP as the means of dosing. 

Three studies used the Northern leopard frog exposing tadpoles of various stages until stage 36 when hind limb development starts. Crump et al. (2002) reported no effects at any concentration up to 200 μg/L on body weight, development, or hind limb emergence. The authors also reported a 7-d LC50 of 577 μg/L. Since the growth and developmental endpoint NOEC were ≥200 and the narcosis endpoint of lethality was 577 μg/L, the mechanism of action on the former endpoints is more likely to be narcosis related.  Croteau et al. (2009) reported only transitory increases in body weight and development at stage 29, which disappeared at stage 34. No linkage was possible of thyroid receptor activation to body weight or development. Hogan et al. (2006) reported LC50 values of 293 and 598 μg/L for stage 26 and 36 tadpoles of the Northern leopard frog. Hogan et al. (2006) also tested wood frog tadpoles using the same procedures and reported a stage 26 LC50 of 153 μg/L. As noted by the authors, possible effects on body weight cannot be discerned due to excessive crowding within test chambers, which can cause stress-related effects on growth. 

A study with the North American bullfrog by Mayer et al. (2003) exposed stage 32 to 36 tadpoles to 0.2 to 20 μg/L OP for 24 hours. Transient earlier sex differentiation occurred at stages 32 to 34, but disappeared entirely by stage 35. No effects on sex ratio from any concentration of OP were found. 

Rohr et al. (2003) exposed the streamside salamander to 5 to 500 μg/L OP for 37 days. Time to hatch, survival, and growth were reduced at 500, but not 50 μg/L OP. 

Overall, effects on amphibians occur in the same concentration ranges of OP as those reported with fish. From the “valid” or “use with care” studies that reported consistent and treatment related effects on amphibians, effects on tadpole development were transient and usually occurred in the same range of concentration causing lethality. Among the studies, there was no linkage of any effect to an estrogenic or thyroid-mediated causation. 

6.3	 Invertebrates

There have been several studies with invertebrates and OP (summarized in attached Table C) that have been scored for reliability as (1) “valid” or (2) “use with care”. Studies scored as either (3) “not valid” or (4) “insufficient information” cannot be used for hazard assessment and explanation for reliability scoring is also given in Table C. The aquatic toxicity of OP has been studied with Daphnia magna, copepods, sea urchins, and snails. 

Two studies with Daphnia magna are shown in attached Table C. Zou and Fingerman (1997) exposed neonates for five days at 10 to 40 μg/L and measured effects on molting. No effects on molting were  reported at any concentration. Huels AG (1992) reported the results of a 21-d life cycle study with Daphnia magna that was conducted according to OECD guideline 202. The study found effects on survival and reproduction at 100, but not 30 μg/L OP. 

With the copepod Tigeropsis japonicus, Marcial et al. (2003) conducted a two-generation test with 0.01 to 10 μg/L OP beginning with nauplii. No consistent and treatment related effects were found in F0 organisms for the endpoints nauplii stage, maturation stage, fecundity, sex ratio or survival. In F1 offspring, developmental delays (nauplii and maturation stages) were noted at most concentrations, but no effects were found on  mature copepods for fecundity, sex ratio or survival (NOEC ≥10 μg/L). 

Arslan et al. (2007) and Arslan and Parlak (2007) reported the results of studies with two sea urchin species Arbacia lizula and Paracentrosus lividus. Due to apparent solvent control effects that were not accounted for in their analysis, the Arslan et al. (2007) study with Arbacia lizula cannot be used as the study was scored as (4) insufficient information. In the study with Paracentrosus lividus (Arslan and Parlak, 2007), sea urchin sperm and eggs were exposed for 72 hours to 5 to 160 μg/L OP.  Developmental effects were found at 20 but not 10 μg/L. A 72-h EC50 for embryotoxicity was estimated from a plot to be 20 μg/L. 

A study with a third sea urchin species, Strongulocentrotus purpuratuns was reported by Roepke et al. (2005). The authors exposed sea urchin eggs to 0.0001 to 5 μg/L OP for 96 hours. An EC50 for developmental effects was reported at 0.174 μg/L. This outlier result is considered unusual and possibly suspect as the corresponding EC50 values for the natural and synthetic estrogens E2 and EE2 from the same study were 14.2 and 30.3 μg/L, respectively. In addition, the EC50 for OP was orders of magnitude more sensitive than other compounds tested by the authors including bisphenol A (EC50 226.5 μg/L). No concentrations of OP or any other of the compounds tested were analytically confirmed. Regardless of the validity of the OP EC50, this study reports results that suggest that developmental effects with this sea urchin species, Strongulocentrotus purpuratuns, are not estrogen-mediated effects. 

There are no reliable (“valid” or “use with care”) studies with mollusks that tested OP. As correctly noted in the Annex XV Report, the studies by Oehlmann et al. (2000) with giant ramshorn snails and dogwhelk and by Jobling et al. (2003) and Duft et al. (2003) with New Zealand mud snail are all properly scored for reliability as (3) “not valid”. All four of the studies suffer from numerous experimental challenges or deficiencies and so cannot be used in a hazard assessment of OP. More details are given in attached Table C. 

	6.4 	Summary of Aquatic NOEC for OP

In conclusion, there are reliable toxicity studies for fish, amphibians, and invertebrates that tested OP. The studies covered all parts of the test organisms’ life cycles from eggs to reproducing adults and cover life stages likely to be sensitive to an endocrine mode of action. Test procedures included screening tests, short-term reproduction tests, and full life-cycle tests. A consistent and treatment-related set of NOEC and LOEC have been reported and range from 10 to 900 μg/L across the apical endpoints related to survival, growth and development, and reproduction. 

7.0 	The Report does not establish that the reproductive and other aquatic toxicity effects of 4-t-OP are solely - or primarily - caused by an endocrine mode of action and an evaluation of the aquatic toxicity of OP does not support concern for “probable serious effects” to the environment that are equivalent to CMR Cat. 1A and 1B, PBT, or vPvB substances.

Relates to Sections 5 and 6 of the Annex Report and to the Report in general

The Report concedes that the question of how to define “probable serious effects to the environment” and how to conclude whether or not they are “of equivalent concern” is still the subject of debate.  While the Report does not suggest any general criteria for these terms, it suggests that the criteria in this case should take into account the potency of the substance and the relevance of the observed effects.  As such the Report develops the case in the OP Annex XV Report around two questions:  
1. Is OP a substance having endocrine disrupting properties?
2. Is there scientific evidence of probable serious effects which give arise to an equivalent concern compared to CMR and/or PBT substances?

Based on the language in Art. 57 it is clear that “equivalent concern” should not rise solely from the mode of action (i.e. endocrine). Rather the effect (i.e. apical endpoint or a population level effect) must be of “equivalent concern” to those of CMR Cat. 1A, 1B, PBT or vPvB compounds.   The Annex XV Report for OP incorrectly suggests that the effects of OP are equivalent to those of known potent estrogens, provides inadequate rationale for a linkage between the very weak estrogenic activity of OP to reproductive or developmental effects in aquatic organisms, and incorrectly attributes obvious general toxicity in amphibians to thyroidal activity. The following comments address these points. 

7.1	OP affects acute lethality and chronic reproduction, growth and development endpoints very differently than natural (E2) or synthetic (EE2) estrogens.

Considerable effort has been made in the Annex XV Report to link effects reported for OP from all the studies with fish to an endocrine disruption mode of action.  Comparison of studies examining OP and natural (E2) and synthetic (EE2) estrogens demonstrates that there are great differences between the effects that they cause to aquatic organisms.

In a life cycle test with zebrafish, Wenzel et al. (2001) demonstrated that estrogenic activity for the natural and synthetic estrogens E2 and EE2 occurs at concentrations that are orders of magnitude below the threshold for systemic toxicity, and result in a suite of responses that are collectively linked to the estrogenic mode of action. In the life cycle study with EE2 and zebrafish, Wenzel et al. (2001) reported the 96-h LC50 (narcosis-based toxicity) for EE2 to be 1700 μg/L and the NOEC for fertilization success to be 0.0003 μg/L. The authors calculated the ratio of the two values to be 5.73 x 106. It is clear for EE2 that the reproduction, growth and development endpoints are affected by a different mechanism of action (endocrine) than the acute lethality (narcosis) endpoints. Given the potency of natural and synthetic estrogens, this is expected, and hence the very high ratio is understandable. 

OP does not affect the same endpoints in aquatic organisms in the same way that natural and synthetic estrogens act. Data from three studies with OP demonstrate that the reproduction, growth and development endpoints are mainly indicative of narcosis-type mode of action, possibly coupled with a very weakly estrogenic mode of action. 

In the life cycle study with OP and zebrafish, Wenzel et al. (2001) reported a 96-h LC50 of 370 μg/L. The ratio of the acute LC50 and the lowest apical development NOEC from this study (fertilization success) of 12 μg/L is 31. In the life cycle study with OP and Japanese medaka, the Japan MOE (2002) did not report an acute value. However, while an LC50 for Japanese medaka is not available for OP, it can be assumed to be about 900 μg/L, based on a review of many data for OP and the structurally similar nonylphenol (NP) that ranged up to 900 μg/L across many freshwater and marine species (Staples et al., 1998). Using the data from the Japanese MOE (2002) report, the ratio of an estimated LC50 of 900 μg/L and the NOEC of fertilization success (30.4 μg/L) was calculated to be 30. For sheepshead minnow, Karels et al. (2003) reported a 72-h LC50 of 720 μg/L for OP, while the NOEC for effects on percent viable eggs was 11.5 μg/L OP. The ratio of the two values is 63. Testing guppy, Toft and Baatrup (2003) reported a 96-h LC50 value of 495 μg/L for OP. A ratio of 50 was calculated using the LC50 and the NOEC of 10 μg/L for the most sensitive reproductive or developmental endpoint, gonopodium length. Thus, the ratio of acute lethality LC50 values and the NOEC of the lowest apical NOEC for reproduction, growth or development endpoints are consistent across the species zebrafish, Japanese medaka, sheepshead minnow, and guppy. Across the four species, the ratios range from 30 to 63, which are approximately 100,000- to 200,000-fold lower than the ratio calculated for EE2. 

Together these data show that OP has a very different mode of action than the natural and synthetic estrogens with effects on survival resulting from narcosis being at least as critical to the organisms exposed to OP as other modes of toxicity. While biomarkers such as VTG indicate that estrogen receptor binding is occurring while exposed to OP, the endpoints related to growth, development, and reproduction are only a factor of 30 to 63 from short term acute toxicity and in the same range as caused by longer term lethality.  Hence it is possible that effects on growth, development, and reproduction for OP are more indicative of narcosis-type mode of action, possibly coupled with a very weakly estrogenic mode of action. Taken together, the acute and chronic datasets for OP and EE2 are very different. 

7.2	The concern expressed in the Annex XV Report on OP is for reproductive or developmental effects in aquatic organisms caused by an endocrine mechanism; however it does not provide adequate rationale for the leap between findings of weak 4-t-OP estrogenic activity and reproductive effects seen in some fish studies. 

The endpoints that are considered to indicate an estrogen agonist mode of action are summarized on page 104 (Table 27) of the Report.  Several of these endpoints are more apical in nature and can be observed naturally and also under various stressful conditions for fish.  Oocyte atresia normally occurs in fish as a part of the natural reproductive cycle, so an understanding of the normal background level of this phenomenon is important. Oocyte atresia can also occur in the ovaries of female fish when exposed not only to an estrogenic compound, but also in response to an androgenic mode of action (Ankley et al., 2003).  Therefore it appears that oocyte atresia, in and of itself, is not indicative of a particular endocrine mode of action.  In addition, a depression of secondary sex characteristics in the fathead minnow or medaka has also been observed in toxicity tests with compounds that operate via a mode of action that is not related specifically to estrogen agonism (USEPA, 2007).  
In various studies with medaka fish, increased VTG was observed in males at concentrations of OP ≥ 11.4 µg/L, altered histopathology of the gonads (specifically occurrence of testis-ova) generally occurred at concentrations of OP ≥ 11.4 µg/L (Gronen et al., 1999; Seki et al., 2003; Gray et al., 1999a; Gray et al., 1999b).  The occurrence of testis-ova in the gonads was more marked when fish were exposed during the time period of sexual differentiation, which occurs before hatch.  Incidences of ova-testis were less frequent and occurred at higher concentrations when exposure to OP was initiated post-hatch (Gray et al, 1999a; Seki et al., 2003).  Male fish with testis-ova were observably undergoing active spermatogenesis and were able to fertilize a female fish, so it appears that the occurrence of testis-ova does not preclude reproduction (Gray et al., 1999b; Seki et al., 2003).  Growth and reproduction were adversely affected at concentrations of OP ≥ 20 µg/L (Gronen et al., 1999; Seki et al., 2003; Gray et al., 1999a; Gray et al., 1999b).  Sex ratios in medaka fish were altered toward a greater proportion of females at concentrations of OP ≥ 48.1 µg/L (Seki et al., 2003; Gray et al., 2009).  As noted by Gray et al. (1999a), effects on apical endpoints that would indicate adverse effects on a population level parameter (such as altered sex ratios, decreased growth and reproduction) occurred at concentrations of OP that are higher than levels found in the environment.  
In guppies exposed for 90 days to nominal concentrations of 1 to 200 μg/L of OP, the lowest concentration to cause statistically significant effects (increased gonopodium length, increased sperm count, decreased ovary weight) was 100 μg/L (Toft and Baatrup, 2003).  This concentration, however, resulted in up to 40% mortality.  The authors stated this concentration of OP was considerably higher than what is found in the environment and concluded “exposure of guppies to environmentally realistic concentrations of OP does not alter the sexual characteristics of these fish”.  When male and female guppies were exposed to 26 μg/L OP for 28 days (males) and 26 to 36 days (females), no differences were observed with survival, gonadosomatic indices, brood intervals, brood size, gonopodium length and sex ratio of the offspring even though there were potential changes to the testis structure (indications of blocked spermatogonial mitosis) in the males (Kinnberg et al, 2003).
In regards to the amphibian data set, the Kloas et al. (1999) study was given a reliability score of (3) “not valid” for several reasons as described in Section 6.2 of these comments. In addition, the Kloas et al. (1999) study should not be considered valid for use with respect to the altered sex ratio endpoint, because gonads of juvenile Xenopus laevis were only inspected based on gross morphology and not on histopathology.  Histopathological examination of the gonads is needed to correctly verify sex, especially in juvenile organisms such as those examined in this study. In addition, due to the non-relevant nature of the administered dose (via injection) and the small sample size, the van Wyk et al. (2003) study is of limited relevance.   In the studies with amphibians that were considered to be valid for use, there were indications of population level effects (on endpoints such as growth, reproduction, survival) at concentrations of OP < 50 µg/L (stage 36).  However, this effect at 50 µg/L (increased body weight) was noted in one stage of Rana pipiens tadpole development and was not noted in other developmental stages with the same species and was not noted among a closely related frog species (e.g., R. sylvatica) in the same experiment (Hogan et al., 2006).  More generally, adverse effects in amphibians were apparent at concentrations of OP ≥ 100 µg/L (Bevan et al., 2003; Rohr et al., 2003), levels that are three orders of magnitude greater than the upper 90th centile concentration of 0.25 µg/L OP in European surface waters.  

In a more recent, well-conducted, chronic study with OP and Xenopus tropicalis, frogs were continuously exposed to 0,  1.1, 3.3, 11, and 36 µg/L OP from Nieuwkoop and Faber stage 46 to adulthood (a total of 31 weeks) with a sampling time-point at the completion of metamorphosis as well (Porter et al., 2011).  In this study, no significant deviation from the control sex ratio was observed for either sampling period, suggesting minimal to no effect of OP exposure on gonad differentiation.  No effects in the adult frogs were observed for mortality, body mass and size, liver somatic index, estradiol and testosterone serum levels, sperm counts, or oocyte counts. The development and growth of oviducts, was observed in males exposed to OP, and VTG levels were increased among juvenile male frogs (sampled at the time of metamorphic completion), but not in adult male frogs at the close of the exposure at 31 weeks.  The transient increase in male vitellogenin and the presence of oviducts in some male frogs exposed to OP indicates weak estrogenic activity of OP, however Porter et al (2011) concludes that the highest OP concentration used, 36 µg/L, was at or below the no observed effect level (NOEL) for general toxicity in X. tropicalis.  

Together, the studies with fish and amphibians did not fully link secondary endpoints such as testis-ova or VTG induction to adverse outcomes. Fish with testis-ova were able to reproduce and male frogs with increased VTG levels developed normally. The Report does not provide adequate rationale for fully connecting findings of weak 4-t-OP estrogenic activity and reproductive or developmental effects seen in some fish studies.

7.3    The case made in the Annex XV Report for OP exhibiting thyroidal activity in
         amphibians is weak and inconclusive, and is not sufficient to warrant OP as being
         of “equivalent concern” as CMR and PBT or vPvB.  

The Annex XV Report states, “4-tert-octylphenol alters the function of the endocrine system in aquatic organisms through an estrogen mediated mode of action, with some evidence of thyroidal activity in amphibians” (pg 6).  However, as will be discussed, the case for OP exhibiting thyroidal activity in amphibians is weak and inconclusive, and is not sufficient to warrant OP as being of “equivalent concern” as CMR and PBT or vPvB.  

The Annex XV Report correctly indicates that the endpoints in the 21-day amphibian metamorphosis assay (OECD Guideline 231) that are intended to indicate potential thyroid activity are: advanced development (measured by advanced developmental stage or increased hind limb length relative to controls), asynchronous development, and/or significant histopathologic alterations of the thyroid glands.  Out of the eight amphibian studies summarized in the Annex XV Report, only studies on two frog species are referenced on page 6 of the Report as providing some evidence for a thyroid agonist mode of action for OP.  These studies include testing on Rana pipens and Rana catesbeiana tadpoles.   These studies will be closely examined for the strength of an OP association with thyroid agonism/potential thyroid activity.

In one study, Rana pipiens tadpoles exposed to OP for eight months showed delayed development and higher body weights (at stage 29 only) starting at 0.02 µg/L OP, increased levels of thyroid receptor β mRNA (at stage 34, but not 29 or 42) at concentrations of 2 µg/L OP, and increased deiodinase III mRNA (at stage 29, but not 34 or 42) at 0.02 but not 2 µg/L OP (Croteau et al., 2009).  Thyroid receptor β binds thyroid hormones and is directly autoinduced by thyroid hormone binding (Denver, 1998; Hogan et al., 2007; Zhang et al 2006).  Deiodinase III inactivates thyroid hormone (Denver, 1998; Hogan et al., 2007).  There were no significant alterations in thyroid hormone levels among stage 29 or 34 tadpoles in this study.  Taken together, the occurrence of increased deiodinase III mRNA at stage 29 (at the low dose of OP), the increased thyroid receptor β mRNA at stage 34 only, the lack of alteration in circulating levels of thyroid hormone levels, and delayed development do not indicate a consistent or cohesive picture of perturbation of thyroid activity in association with OP exposure in the Croteau et al. (2009) study.  Specifically, increased thyroid receptor β mRNA should correspond with increased thyroid hormone levels and advanced, not delayed, development..  Another consideration for this particular study with R. pipens is that the tadpoles were reared at relatively high densities, which may have resulted in stress to the tadpoles potentially interfering with measured endpoints (e.g., increased rearing density increases corticosterone (Glennmier and Denver 2002a), which can slow growth and development (Glennmier and Denver, 2002b).  Also, no positive control was used in this study design to establish what responses would be expected for a known thyroid active compound in this species with the selected endpoints.  In another study with R. pipens tadpoles, shortened time to hind limb emergence (advanced development) was observed among tadpoles exposed to 200 µg/L OP and ultraviolet B (UVB) radiation for 10 days.  This same effect was not noted when tadpoles were only exposed to OP (and not UVB) for 10 days (Crump et al., 2002).  Therefore, there was no indication of thyroid agonism for OP acting alone, apart from UVB radiation (Crump et al., 2002).    

 In another study, Rana catesbiena tadpoles were exposed to OP for 24 hours and an earlier completion of sexual differentiation was observed at concentrations of OP ≤ 0.2 µg/L (Mayer et al., 2003).  However, there is no definitive link between an earlier completion of sexual differentiation and thyroid agonism and/or thyroid perturbation.  The process of gonad differentiation in amphibians appears to be controlled separately from activity in the hypothalamus-pituitary-thyroid axis (Robertson and Kelley, 1996; Buchholz et al., 2006; Coady et al., 2010).  Therefore, this study should not be used to support the potential thyroid agonsim of OP.  Furthermore, in a recent well-designed chronic study wherein the western clawed frog (Xenopus tropicalis) was exposed to four concentrations of OP (ranging from 1 to 36 µg/L) beginning at premetamorphic developmental stages and lasting until sexual maturity (31 weeks), there were no effects on gonad differentiation in OP exposed individuals relative to controls (Porter et al, 2011).  Thus, there are no definitive data to indicate that OP acts on the thyroid system of amphibians either as a thyroid agonist or antagonist, and this line of evidence should not be used to indicate that OP has endocrine disrupting characteristics that are of equivalent concern to CMR, PBT or vPvB classification status.

In summation, results of aquatic toxicity tests conducted OP and EE2 show that: 
1) effects on acute lethality, reproduction, growth, and development are very different for the two substances; 
2) studies with fish and frogs exposed to OP failed to fully link an endocrine mode of action affecting secondary endpoints with adverse reproductive, growth or developmental outcomes; and 
3) there are no definitive data to indicate that OP acts on the thyroid system of amphibians either as a thyroid agonist or antagonist. Together, these data represent further scientific evidence that OP does not cause “probable serious effects” to the environment that are of equivalent concern to CMR Cat. 1a and 1b, PBT, or vPvB substances.

8.0   	The arguments listed in the summary of the Annex XV Report on OP are not supported by an adequate scientific basis to propose identifying this compound as a Substance of Very High Concern (SVHC) on the basis of “equivalent concern” for endocrine modulated environmental effects.

Relates to the Summary and Section 6 of the Annex Report and to the Report in general

The Annex XV Report states that OP “is proposed to be identified as a “substance of equivalent concern according to Article 57 (f) because of its environmental endocrine disrupting properties and subsequent serious effects to the environment” and lists a series of arguments or concern as a basis for this proposal. However, these arguments are not supported when considered relative to the data provided previously in these comments. The following comments respond to the arguments listed as the basis for this proposal.   

8.1	 The case made in the Annex XV Report for OP exhibiting thyroidal activity in amphibians is weak and inconclusive, and is not sufficient to warrant OP as being of “equivalent concern” as CMR and PBT or vPvB.  
The Report summary states: 
•	tert-octylphenol alters the function of the endocrine system in aquatic organisms through an estrogen mediated mode of action, with some evidence of thyroidal activity in amphibians”  
•	 Effects that occurred in two other amphibian species (R. catesbeiana and R .pipens) provide some evidence for a thyroid agonist mode of action. Both effect types indicate that 4-tert-octylphenol might be an endocrine disruptor in amphibians
As discussed in Section 7.3 of these comments, the case made in the Annex XV Report for OP exhibiting thyroidal activity in amphibians is unsubstantiated. Therefore, there is no basis to support the above summary statement and the data are not sufficient to warrant OP as being of “equivalent concern” as CMR and PBT or vPvB.  

8.2 	Adverse effects in fish species are not “clearly endocrine mediated”; rather they are more indicative of general toxicity possibly coupled with very weakly estrogenic activity. 

The Report summary states: 

•	Adverse effects observed in three fish species are clearly endocrine mediated
•	In all other tested fish species the endpoints affected are known to be influenced by estrogen activity and, with only one exemption, an estrogen mediated mode of action was observed.
•	Ranges of NOEC were essentially the same for survival, growth and development, and reproduction endpoints

The first summary statement listed above refers to studies with Japanese medaka, sheepshead minnow, and guppy fish. The following table (Table 5) shows that more than half of the NOEC for these species were actually "no effects at any concentration of OP tested". The range of concentrations showing effects in fish was about 20 to 900 µg/L OP.

See TABLE 5  Summary of effects from exposure to octylphenol for three fish species in attached comments 

As discussed in Section 5.0 of these comments, there were issues with study validity scoring and interpretation in the assessment of fish data in the Annex XV Report.  The endpoints showing effects (survival, growth, and reproductive endpoints) are not necessarily "endocrine mediated". Interpreted properly, the data show that the ratio of acute lethality (LC50) to the lowest reproductive NOEC for OP were 30 to 63 for medaka, sheepshead minnow, and guppy. A similar ratio of 31 for zebrafish is also reported. The comparable ratio for the potent synthetic estrogen EE2 with zebrafish is 5,700,000. The vast difference in these ratios show that effects caused by OP are not "clearly endocrine mediated", but rather are indicative of general toxicity possibly coupled with very weakly estrogenic activity. 

8.3   Effects observed for studies of OP in amphibian species are not comparable to
        those observed for 17ß-estradiol.
     
The Report summary states: 
•	the effects observed in the amphibian species X.laevis after exposure to 4-tert-octylphenol are comparable to effects observed for 17ß-estradiol 

 As discussed in Section 6.3 of these comments, two of the three studies cited with this species were scored (3) "not valid" and are not reliable (see previous comments and attached Table B). Kloas et al. (1999) was found to have numerous experimental design flaws. Van Wyk et al. (2003) utilized intraperitonial injection of the frogs. The third study only reported effects with OP with effects on growth and development at 100 and 1000 µg/L OP, respectively; however there was no positive control and therefore no basis for comparison to E2. Therefore, there is no basis to support the summary statement above that effects observed for OP in amphibian species are comparable to E2. 
8.4	The Annex XV Report does not adequately support its argument that OP is a potent endocrine disruptor 
The Report summary states: 
•	4-tert-octylphenol is a potent endocrine disruptor with regard to the concentration required to cause endocrine mediated effects and the exposure duration needed to cause effects  
•	Tests with some fish species testing 17ß-estradiol and 4-tert-octylphenol indicated that 4-tert-octylphenol may be nearly as potent as the natural estrogen 17ß-estradiol (factor 2- 28 less potent)

As discussed in Section 3.0 of these comments, the Report does not adequately support its argument that OP is a potent endocrine disruptor. The summary statements listed above are simply not supported by the available data.   The relative potencies for OP relative to E2 or EE2 cited in the Annex XV Report (2 to 28) were not calculated correctly.  More than half of the fish and amphibian NOEC were actually no effects at any concentration of OP tested. The range of concentrations showing effects was about 20 to 900 µg/L OP. The endpoints showing effects included survival, growth, and reproductive endpoints and NOEC for these end points are all similar.  When properly calculated from reliable studies, the estrogenic potency of OP is in the range of 1000 to 40,000 less potent than E2 or EE2 based on numerous in vitro and in vivo studies. 

Ratios of acute lethality to lowest reproductive effect NOEC (acute-to-chronic) are 30 to 63 for OP, as compared to 5,700,000 for EE2. OP effects are more likely caused by general toxicity possibly coupled with very weak estrogenic activity. Studies with fish have been conducted with exposure regimes that cover all times of the life cycle from egg to embryo to juvenile to reproducing adult. All critical windows of exposure during any stage of fish development have been tested for effects from OP. 

8.5	The Annex XV Report does not adequately support its argument that OP has effects like a potent estrogen 

The Report summary states: 
•	Taking our knowledge on potent estrogen agonists into account, the level of concern for 4-tertoctylphenol seems to be comparable to PBT /vPvB and CMR substances. 
•	Estrogens are known to result in irreversible, persistent effects.

Section 4.0 of these comments examines the estrogenic potency, efficacy and aquatic toxicity of natural estrogen (E2) and synthetic estrogen (EE2) and provides an understanding of what constitutes a highly potent estrogen.  The effects on aquatic organisms by potent estrogenic substances such as E2 and EE2 collectively add up to a predictable, cohesive signature of effects, which are consistent with the known mechanism of action for these compounds. Natural development of male gonadal tissue is halted, differentiation into male characteristics is blocked, male secondary sex characteristics are prevented from developing, and gonadal tissues are severely compromised and damaged, reproduction within natural populations can be halted, and effects that are mediated by estrogenic activity occur at concentrations that are millions of times lower than those where narcotic effects occur.  In Section 6.0 of these comments a scientifically rigorous review of relevant studies shows the effects of OP on aquatic organisms are very different from the effects from potent estrogens and are not comparable to PBT/vPvB or CMR Cat. 1A or 1B compounds. 

8.6	Sufficient studies covering all critical windows of exposure during all stages of fish development have been considered in the aquatic studies for this compound.

The Report summary states: 

•	A change in the endocrine feedback system during sensitive life stages may result 
             in effects during the entire life. 
•	Impairment of reproduction may already occur after a transient short term exposure of adults as observed in two fish species (21d exposure of adult males)

As discussed in Section 6.0 of these comments, fish have been tested with OP over all critical windows of exposure and life stages likely to be sensitive to endocrine modes of action with a NOEC range from about 10 to 900 µg/L OP. The duration of these studies ranged from short term exposures to life-cycle exposures and the NOEC and LOEC values that have been developed account for any life stage sensitive effects. 

8.7	OP does not meet the bioaccumulative criteria for SVHC compounds and its “distribution toward embryos in viviparous species” did not cause developmental effects in offspring or raise concern for the maternal transfer of OP within female guppy fish.

The Report summary states “with respect to 4-tert-octylphenol the level of concern is increased by its bioaccumulation potential with some evidence of internal distribution toward embryos in viviparous species.

This supporting statement is misleading and inconsistent with the conclusion in section 3.3 of the Annex XV Report that OP is only low to moderately bioaccumulative (as evidenced by laboratory BCF results ranging from 46 to 471) and does not meet the bioaccumulation criteria to be considered a SVHC. The observation "internal distribution toward embryos in viviparous species” was not attributed to a specific study in the Report summary, but may relate to a study by Kinnberg et al. (2003) in which the authors did not find any "level of concern" related to maternal transfer of OP within female guppy fish.  The authors exposed female guppy (a viviparous species) to 26 μg/L OP or 0.85 μg/L E2. According to the authors, "At the tested concentrations, exposure to octylphenol or 17B-estradiol via the mother fish did not significantly affect the weight, length, gonopodium index or sex differentiation of the offspring". In addition, a study with the viviparous species eelpout (Rasmussen et al. (2002) examined the offspring from a female fish exposed to 0, 25, or 100 μg/L OP. Significant effects on the offspring were only found in offspring from the females exposed to 100, but not 25 μg/L OP, including sexual differentiation and VTG induction. These results are fully in line with the range of NOEC and LOEC across numerous species, do not show any unexpected results and do not warrant equivalent concern to a CMR Cat. 1A or 1B or a PBT/vPvB compound.  

8.8 	OP does not meet persistence criteria for SVHC compounds. 

The Report summary states that OP “is persistent under anaerobic conditions combined with a tendency to adsorb to sediment and soil”.   

This supporting statement is misleading and inconsistent with the conclusion in section 3.1 of the Annex XV Report. OP does not meet any of the persistence criteria for SVHC compounds. As is true for many organic substances, OP degrades more slowly under anoxic conditions. The moderate sorption capacity to soil and sediment acts to keep OP from being transported further after release into those compartments. 

8.9	Considering the low estrogenic potency of OP, along with its low occurrence and concentrations in the environment, it is not likely, or probable, that this compound will result in effects that are adverse to that ecosystem with respect to community structure and function. 
  
The Report summary states:
•	Population relevant endpoints affected in these species are known to be sensitive toward estrogen activity, although not being diagnostic for this mode of action  
•	Exposure to 4-tert-octylphenol may result in effects that relevantly influence ecosystems with respect to the community structure and function
•	Comparable to other estrogens, 4-tert-octylphenol influences reproduction parameters as well as sexual development (including changes in sex-ratio) and growth and thus endpoints are affected that may impair population stability and recruitment.
•	Several species belonging to different taxonomic groups were affected at low concentrations indicating that 4-tert-octylphenol may cause endocrine mediated effects in a variety of species.
•	Some evidence for potent estrogen agonists is available that population stability is affected even after exposure has ceased. 
•	Due to a highly conserved receptor among vertebrate species it is likely that several fish species representing a variety of reproduction strategies and migration patterns may be affected.
•	The type of effects observed (e.g., short exposure to 4-tert-octylphenol in adults affects fertilization success) provide some evidence that exposure in one area might influence population stability in another area (e.g., for salmonids migrating long distances passing highly polluted areas during migration towards spawning grounds). 

The data provided in these comments shows that OP has only weakly estrogenic activity, probably combined with general toxicity (e.g, narcosis) and is clearly not "comparable to other estrogens".  As noted above, the potency for OP relative to E2 or EE2, when properly calculated from reliable studies, is in the range of 1000 to 40,000 fold less potent across numerous in vitro and in vivo studies.  Adverse apical effects caused by OP are not "clearly endocrine mediated" but rather are indicative of general toxicity possible coupled with very weakly estrogenic activity.   All valid NOEC based on apical endpoints ranged from about 10 to 900 µg/L. Concentrations measured in Europe were reported to be 0.05 μg/L (median concentration) to 0.25 μg/L(upper 90th percentile concentration), showing that concentrations were well below the full range of NOEC for OP so the probability of OP causing population level effects is low. 	Thank you for your specific comments. As described above they were carefully considered in detail. Some details were changed in the dossier, but this did not change the overall conclusion.
 
2.0 – 2.2 We appreciate your analysis. However, you might be aware, that the dossier does not aim at suggesting OP as CMR Cat 1A or 1B PBT or vPvB substance.

3.0 Thank you for your comments please consider our response to comments below. 

3.1 The relative in vitro potency has been included in chapter 6.2.1.2. Depending on the fish species relative potency differs between > 0.001 (C.carpio and Z.viviparous) and < 0.00009 (S.salar, O.mykiss). 

3.2 A more detailed description of the in vivo relative potency compared to 17ß-estradiol has been included in chapter 6.2.1.2. 
Only apical endpoints were considered that are likely to be influenced by the endocrine mode of action (see also comments below) Please note, that not NOEC but LOEC values were reported in the tables and that it was always indicated if no effects were observed up to the highest test concentration (indicated as LOEC > highest tested concentration). Comparison of LOEC values show, that, as indicated in this comment, relative potency for some fish species is low (relative potency  0.00003 – 0.000005). However, results for other species indicate that relative in vivo potency might be higher (0.5 – 0-007 for O.latipes, Z.viviparous and P. Reticulata). We agree that these data should be used with care due to deficiencies of studies. However, please note that data were included for information only as the overall weight of evidence conclusion on the SVHC status of OP is not based on the relative potency compared to 17ß-estradiol

3.3 See also comments above. Please note, that a full comparison is possible for Z.viviparous only. In this case relative potency in the in vivo study was factor 10 higher than in the in vitro tests (0.01 compared to 0.001). Results for O.mykiss cannot be compared since in vitro E2 was tested while EE2 was tested in vivo. 

.

4.0 We appreciate your comment and your careful analysis of effects observed for E2 and EE2. If compared to effects observed for OP, it becomes obvious that the types of effect are very similar (complete feminization of male gonads; induced failure to differentiate into males; reduced reproduction capabilities; severe histological effects on gonadal tissues) but higher concentrations of OP are needed to cause these effects. 

5.0 Thank you for your comment Your analysis was carefully checked. Please see our response to comment 5.1 – 6.4 with regard to the validity of studies, our study interpretation and scientific analysis.

5.1 All deviating Klimisch codes were reassessed carefully (see more specific response to comments 6.1 – 6.3)

5.2 The dossier was revised and the phrase “starting at” was removed. However, we would like to emphasize that – with the exception of P.reticulata - all conclusions were based on statistically significant effects. Effects for which no statistics was provided remained in the dossier as they provide additional information on the mode of action in a weight of evidence approach.

Thank you for your comment. Please find our responses to comments 6.1 – 6.4 with respect to the validity of studies and their scientific analysis. Throughout yourcomments it is stated that effects on systemic endpoints such as Reproduction, growth or mortality are indicators for a narcotic endpoint rather than providing evidence for an estrogen mode of action. We agree that effects on systemic endpoints do not provide information about the mode of action (neither with regard to specific modes of action nor with regard to narcosis). However, if information from different levels of information (in vitro information, biomarker, histological changes, comparison of effects with effects caused by E2 and EE2) is considered, some evidence about the mode of action is provided.

6.1: with respect to fish, deviating Klimisch code scores were proposed by CEPAD for 8 studies (four with major changes from 1 or 2 to 3 or 4). All these studies were carefully reassessed: 
 
Medaka, Gray et al 1999b (note that this study is indicated as Gray et al 1999a in Table B of your comment): reliability remained 2 as the main experiment (behavior after cessation of exposure) started after mortality has been observed. 10 replicates were used for this experiment

Medaka, Knörr and Braunbeck, 2002: Reliability remained 2 as statistic was provided for some endpoints. If no statistics was provided, this was clearly marked in table 28.

Guppy, Bayley et al 1999: reliability remained 2 as the main experiment (behavior after cessation of exposure) started after mortality has been observed. 15 replicates were used for this experiment.

Guppy, Japanese Ministry, 2002: Score was transitionally changed from 2 to 4 as documentation is not available yet

Please note, that none of these studies were decisive for the overall conclusion All results were supported by other studies in a weight of evidence.

NOEC ranges described in your comment seem not to fit to those reported in the dossier (which reports LOEC values) but this is not discussed in detail as the exact NOEC values are not considered relevant for the overall conclusion. 

6.2 Amphibians

Reliability of the two studies, with deviating Klimisch code scores, was reassessed carefully:
Van Wyk et al 2003: scoring was not revised as an unrealistic dosing method does not change reliability but indicates lower relevance. 
The score for the study by Kloas et al 1999 was revised from 2 to 3 in line with your comment. Please note that this did not change the overall conclusion as information for amphibians was used as supporting information only. The dossier was revised in order to state this more clearly.

Results summarized in your comment match to those summarized in table 42 of the SVHC dossier. However, data provided in table 42 clearly indicate that effects on development start well below concentrations causing lethality. Although there is no clear link between effects and a specific endocrine mode of action, comparison with effects observed for 17ß-estradiol provide some hints that observed adverse effects might be endocrine mediated. The Dossier was revised on order to state more clearly that information does not prove such a link. 

6.3 Invertebrates: 
Reliability of the three studies with deviating Klimisch code scores were reassessed carefully.
Reliability with respect to the study by Andersen et al, 2001 was revised according to your comment from 1 to 2. No revision was made for the other two studies (Huels AG  1992 remained score 4 due to missing documentation and Arslan et al 2007 remained score 2 as the study is considered well documented and effects are correctly compared to the solvent control)…  
Please note, that none of the studies was decisive for the overall conclusion.

Results summarized in your comment match to those summarized in table 42 except for one study. Effects reported by Arslan et al (2007) for P.lividus 
resulted in a LOEC of 5 µg/L with respect to larval malformations after exposure of sperms and eggs respectively. With respect to T. japonicas (Marcial et al, 2003) it should be noted, that effects observed in F1 offspring results in a LOEC of ≤ 0.01 µg/L.
With respect to mollusks, the dossier was revised on order to state more clearly that studies are of low quality and provide some hints only.

7.0 and 7.1  Please consider chapter 5.1.2.3 fish (Table 29, 30, 32, 34, 36,38 and 40 and the corresponding text) where available information is compared to guidance provided in the OECD draft guidance document for evaluating chemicals for endocrine Disruption. Results show, that at least for one fish species OP should be considered as an actual endocrine disruptor while for several other fish species there is strong evidence. With regard to amphibians and invertebrates no such conclusion was drawn in the dossier but some information suggest the possibility of endocrine mediated adverse effects. The dossier was revised in order to state this more clearly. 

We appreciate your comment. It suggests that due to differing ACR values, the mode of action may not be similar to E2 or EE2. However ACRs for OP are well above narcotic ACRs. Lower ACRs compared to E2 and EE2 may provide some information about the potency but in our opinion they do not provide information about the actual mode of action. As described in chapter 5.1.2.3, type of effects observed after exposure to OP are in general very similar to those observed for E2 or EE2.

7.2 See comment above (7.0). Please note that endpoints summarized in table 27 were derived from (draft) OECD guidelines. However, neither oocyte atresia nor depression of secondary sex characteristics were decisive for the overall conclusion.
Please also note, that histological changes and vitellogenin induction were not used as apical endpoints but were reported as supporting information providing evidence that apical effects are linked to an endocrine mode of action Please also note, that the SVHC identification is based on the intrinsic endocrine disrupting properties causing adverse effects and that exposure concentration is irrelevant for identification.
The dossier was revised on order to state more clearly that information available for amphibians does not prove a clear link between effects observed and an endocrine mode of action. 
Thank you for the information about the additional study by Porter et al (2011). Without having analysed the study in detail it seems that it supports an estrogenic mode of action in amphibians.

7.3 The overall conclusion in section 5.1.2.4 was slightly rephrased in order to state more clearly that there are only some indications of a possible endocrine mode of action  with regard to amphibians (see also response to comment 16 below).

8.0 Thank you for your comment, see response to comments below

8.1 .Chapter 5.1.2.4 was revised. The overall conclusion was rephrased in order to stage more clearly that there are only some indications of a possible endocrine mode of action with regard to amphibians (see also response to comment 16 below). This does not change the overall conclusion.

8.2 see response to comment 6.1. Chapter 5.1.2.3 summarizes the evidence that adverse effects are endocrine mediated. Please note that summary of effects in table 5 of your comment does not fit to data provided in table 28 for O.latipes. Even if only data provided in studies that were considered reliable 1 or 2 by CEPAD were used, effects on fertility and fecundity, sex-ratio, development and embryo survival occurred at 82.3 µg/L and below. If all studies which are considered reliable in the dossier are included, some results indicate that effects may start at ≤ 2 µg/L (no statistics). With regard to P.reticulata statistics are mainly missing. However data (scored as 1 or 2 by CEPAD) provide some evidence, that effects on reproduction and courtship behaviour start at ≤ 1.7 µg/L

8.3 .Chapter 5.1.2.4 was revised. The overall conclusion rephrased in order to stage more clearly that there are only some indications of a possible endocrine mode of action with regard to amphibians (see also response to comment 16 below). This does not change the overall conclusion.

8.4 see response to comment 3.1- 3.3.with regard to the potency and response to comment 8.2 with regard to the effect concentration values. Results for some fish species indicate in vivo potency of 0.5 – 0.007 compared to E2. The overall conclusion is not based on the relative potency compared to E2. As described above, in our point of view ACR do not provide information, that the actual mode of action is not endocrine.

8.5 see also response to comment 4.0. As is indicated in the dossier, effects described for E2 fit to those observed for 4-tert-octylphenol, but higher concentrations are needed to cause these effects.

8.6 as described in the comment, all sensitive life stages are covered for fish and thus most effects that may occur are covered by the tests. Sentences cited in the comment did not aim at stating that tests are missing but rather state, that even after short term exposure of wildlife, populations may be adversely effected during the whole life time of individuals exposed. 

8.7 The sentence referred to a study by Rasmussen et al (2002) for the eelpout. A significant reduction in weight and body length was observed in offspring exposed via the mother at the lowest test concentration while no adverse effects were observed for adults (LOEC ≤ 14 µg/L). Measured OP concentrations indicated that OP did accumulate in the maternal plasma (BCF 201 and 546 for 14 and 65 µg/L respectively) and was transferred to the ovarion fluid (BCF 0.2 and 15 respectively)

8.8 Please note that the dossier does not aim at identifying OP as PBT substance. However, slow biodegradation adds to the overall concern even if the P criterion is not met.

8.9 With regard to potency and effects comparable to E2 see also response to comment 4.0 and 8.5.: As is indicated in the dossier, effects described for E2 fit to those observed for 4-tert-octylphenol, but higher concentrations are needed to cause these effects. 

With regard to the question whether apical effects are endocrine mediated see also response to comment 6.1, 7.0 and 8.2. Chapter 5.1.2.3 of the dossier clearly summarizes the evidence that adverse effects are endocrine mediated.
	
			See also:
Comment 20 attachment (CEPAD-APERC Comments on OP SVHC (Oct. 13, 2011).zip).zip		
19	2011/10/13	Industry or trade association
Alkylphenols & Ethoxylates Research Council
United States	2.0	The properties of OP fall well short of the Article 57 criteria for CMR Cat. 1A or 1B, PBT, vPvB.  

Relates to Sections 4, 5 and 6 of the Annex XV Report

2.1	The Annex XV Report concludes that OP does not meet the criteria for probable serious effects for SVHCs as defined under CMR Cat. 1A or 1B; it also does not meet criteria for the weakest category, CMR Cat. 2.

Criteria to determine whether a substance should be identified as a “CMR Cat. 1A or 1B” or Cat. 2 are given in the EC  Regulation No 1272/2008 (EC, 2008, Dec. 16) on classification, labeling and packaging of substances and mixtures.  Under this regulation, “reproductive toxicity includes adverse effects on sexual function and fertility in adult males and females, as well as developmental toxicity in the offspring”. The subcategories for reproductive toxicants and their respective criteria are defined under EC Regulation No 1272/2008 as: 

Category 1A (Known Human Reproductive Toxicant), which is largely based 
         on evidence from humans.  

Category 1B (Presumed Human Reproductive Toxicant), which is largely based 
                      on data from animal studies.  

EC Regulation No 1272/2008 requires that “the data shall provide clear evidence of an adverse effect on sexual function and fertility or on development in the absence of other toxic effects, or if occurring together with other toxic effects the adverse effect on reproduction is considered not to be a secondary non-specific consequence of other toxic effects. However, when there is mechanistic information that raises doubt about the relevance of the effect for humans, classification in Category 2 may be more appropriate.

Category 2 (Suspected Human Reproductive Toxicant), which is intended  for substances where there is some evidence from humans or experimental animals, possibly supplemented with other information, of an adverse effect on sexual function and fertility, or on development, and where the evidence is not sufficiently convincing to place the substance in Category 1. 

If deficiencies in the study make the quality of evidence less convincing, Category 2 could be the more appropriate classification. Such effects shall have been observed in the absence of other toxic effects, or if occurring together with other toxic effects the adverse effect on reproduction is considered not to be a secondary non-specific consequence of the other toxic effects.

Substances that are CMR Cat. 1A and 1B are of concern because there is sufficient data to determine that they are “known” or “presumed to be” carcinogenic, mutagenic or toxic to reproduction. Carcinogens induce or increase the incidence of cancer. Mutagens cause a permanent change in the amount or structure of genetic material in cells.  Reproductive toxicants cause adverse effects on sexual function and fertility or on development off offspring in the absence of other toxic effects. 

2.2	  OP is not a PBT or a vPvB

A PBT or vPvB substance is one that meets the clear definitions according to Annex XIII of Regulation (EC) No 1907/2006 and to the guidance on information requirements and chemical safety assessment Chapter R.11 (PBT Assessment, ECHA (2008)).  The European Center for Ecotoxicology and Toxicology of Chemicals (ECETOC) Technical Report on the Risk Assessment of PBT Chemicals states: 

 “chemicals categorized as meeting the P or vP criteria are of particular concern because of their predicted long environmental half-lives. As a consequence, successive releases over time could result in a build-up in the environment if the rate of release is higher than the rate of disappearance. Additionally, when distribution within the environment occurs, the presence of such chemicals could be observed on a wider geographical scale, therefore spreading the possible concern to remote areas. Moreover, when a toxic substance is not only persistent in the environment, but also able to bioaccumulate, the potential for exposure to living organisms in a higher trophic level to a particular substance is increased. This may lead to possible adverse sublethal effects from chronic exposure that would not have been predicted from standardised laboratory tests (ECETOC, 2006)." 

While there are many substances that may be persistent, bioaccumulative, or toxic, it is the unique combination of all three properties that defines the overall hazard potential of a PBT. The threat posed by a PBT substance is that it enters the environment, does not degrade, cannot be metabolized by the organisms that ingest it, and poses toxicity to those organisms. A substance designated “very persistent and very bioaccumulative, or vPvB, have degradation properties that are even slower than for meeting the persistence criteria for a PBT substance. In addition, vPvB substances have even greater bioaccumulation criteria than for a PBT substance and  are considered to be so persistent and bioaccumulative, more so than a PBT substance, that they are treated in the same regulatory manner as CMR and PBT substances, regardless of any toxicity concerns. 

2.2.1	Persistence Assessment (P)

While the Annex XV report includes some discussion of degradation data for OP in these media, a more complete assessment of the expected environmental persistence in alignment with the REACH submission for OP (IUCLID, 2010) is given below. 

According to Annex XIII, a substance does fulfill the criteria “persistent (P)”, if: 
-      the half-life in marine water is higher than 60 days, or 
-      the half-life in fresh- or estuarine water is higher than 40 days, or 
-      the half-life in marine sediment is higher than 180 days, or 
-      the half-life in fresh- or estuarine water sediment is higher than 120 days, or 
-      the half-life in soil is higher than 120 days. 

Simulation tests for the biodegradation of OP and degradation intermediates are available for all environmental compartments listed above except for soil, which can be addressed using read-across from the analogue substance nonylphenol (NP).  Half-lives of OP biodegradation in the different environmental compartments are summarized in Table 1. The available data demonstrate OP is biodegradable in river bed sediments, marine sediments, surface waters, and soil under aerobic conditions. Furthermore the data support a conclusion that OP does not meet the criteria for persistency in any of these environmental compartments. 

While OP was found to not significantly biodegrade under anaerobic conditions, many organic compounds degrade slower under low temperature, nutrient-poor or oxygen-limited conditions (see for example Boethling and Mackay, 2000). The data for OP are consistent with the findings for other such substances.  OP does degrade in the environmental compartments in which organisms reside. These compartments include freshwater and marine water and sediment and soil. Because OP does degrade in these compartments, it is not expected to  build up in the environment and thus does not pose the concern caused by truly long lasting and persistent substances. Therefore, OP is not considered “persistent” or “very persistent” according to Annex XIII criteria.

See Table 1. Summary of Aerobic Biodegradation Data for Octylphenol (Provided in attached comments document) 

2.2.2 	Bioaccumulation Assessment  
The Annex XV Report presents a useful summary of the available data on the bioaccumulation potential of OP. The report gives a range of experimentally determined bioconcentration factors (BCF) of 46 to 471. 
According to Annex XIII, a substance does not fulfill the criteria of “bioaccumulative (B)” or “very bioaccumulative (vB)” if the BCF are below 2000 or 5000, respectively.  The EU Commission reviewed OP in the course of “Interim Strategy for Management of PBT and vPvB Substances” and confirmed that this compound does not fulfill PBT/vPvB criteria (ECB, 2006).  In the Annex XV report for OP, the stated conclusion is that the bioaccumulation potential of OP is low to moderate. 
Overall, the weight of evidence for OP metabolism suggests a low potential for bioaccumulation due to rapid elimination via liver hydroxylation, glucuronidation, and sulfation. Bioaccumulation is only observed if these detoxification pathways are saturated by excessive doses  (IUCLID, 2010) 
The concern posed by bioaccumulative substances that meet the PBT criteria is that they may have the potential to also biomagnify (build up in food-chains and cause increasing exposure with higher trophic levels, including humans, aquatic mammals, and birds). As the studies cited above show, and as noted in the ECB (2006) review, mammals and fish are capable of rapidly metabolizing OP; therefore this compound has low to moderate bioaccumulation potential, which eliminates the concern of increasing accumulation within food webs that are relevant to compounds that meet the PBT or vPvB criteria.  
In summary, OP is neither a “bioaccumulative” or “very bioaccumulative” substance according to Annex XIII criteria.
3.0	The Report does not adequately support its argument that OP is a potent endocrine disruptor. 
Relates to Sections 4, 5 and 6 of the Annex Report and to the Report in general

3.1 	OP is considerably less potent than the natural estrogen, 17β-estradiol (E2) and synthetic (EE2) estrogens based on in vitro estrogen activity screening tests.

Valuable information about the relative potency of endocrine active compounds relative to the known potent estrogens E2 and EE2 are provided by various in vitro screening tests.  As summarized in the table below (Table 2), relative potencies of either E2 or EE2 and OP ranged from 0.000032 to 0.0024 (400 to 31,000 fold less potent) for vitellogenin (VTG) induction and estrogen receptor binding in hepatocytes from several species of fish. As an example, using an in vitro assay with rainbow trout hepatocytes, Jobling and Sumpter (1993) reported a relative potency of OP to E2 of 0.000037, or 27,000 times less potent than E2

3.2	OP is also considerably less potent than E2 and EE2 based on in vivo studies that examined endocrine-sensitive apical endpoints.  

The Annex XV Report greatly overstates the estrogenic potency of OP and goes to great lengths to calculate effect ratios for various endpoints from studies with OP and E2 or EE2 in an attempt to assess its potency relative to these highly estrogenic compounds.  It should be noted that apical endpoints reflect the integration of multiple different modes of action at the biochemical level; therefore calculations of the relative estrogenic potency of compounds based on No Observed Effect Concentrations (NOEC) for apical endpoints does not necessarily provide a specific measure of estrogenic potency.   In addition to this general issue, the following comments raise other concerns about the approach taken in this section of the Report. 

Some of the NOEC ratios presented in the Annex XV Report are not at all useful because they were determined from studies that used only one or two concentrations of either the hormone or OP. In other words, the Report compares NOEC for an endpoint in studies where there were no effects in the study (i.e., no reported LOEC) to a NOEC that did have an LOEC.  This approach is not scientifically founded and provides no useful information about the estrogenic potency of OP relative to E2 or EE2.  For OP, roughly half of all NOEC reported in the Report were actually “no effects at the highest concentration tested”; therefore the actual NOEC could be much higher.  It is inappropriate to represent such results as meaningful for E2 or EE2 to OP comparisons. 

If there is interest in comparing the relative potency of two compounds based on NOEC data for apical endpoints, the most meaningful approach is to compare NOEC from studies where an LOEC was found based on dose-response.  The best examples of this are from the life cycle studies with EE2 and OP and zebrafish conducted by Wenzel et al. (2001). The tests with EE2 and OP were done nearly identically. Numerous identical endpoints were recorded.  NOEC for each endpoint that also recorded an LOEC for both EE2 and OP were F0 juvenile length, F0 time to spawn, F0 eggs per female per day, and F0 fertilization success. The NOEC ratios for OP:EE2 are 0.000025 (or 40,000-fold less potent) for each endpoint. If the ratios of the LOEC for the same endpoints are used, the ratios are 0.000031 (or 32,000-fold less potent). Thus EE2 is 32,000 to 40,000 times more potent than OP, based on the most important endpoints from the full life cycle study. 
  
Relevant studies of shorter term duration confirm these relative potency results (see summary Table 2 below). Using NOEC that are consistent and treatment related, findings from the study with guppies by Toft and Baatrup (2003) showed that estrogenic hormones were 2,000-fold more potent than OP in regards to sex ratio and 1,000-fold more potent than OP in regard to length and colorization. Van den Belt et al. (2001) showed that estrogenic hormones were 20,000-fold more potent than OP for percent spawning and 2500-fold more potent than OP for ovarian-somatic index (OSI). 

3.3	Relative potencies for OP:E2 or OP:EE2 are similar with in vitro and in vivo testing

Relative potencies are shown in the summary table (Table 2) for in vitro and in vivo tests using studies on E2 or EE2 and OP for which direct comparisons of effects could be made. There is little difference between the range of in vitro (400 to 32,000-fold) and in vivo relative potencies (1000 to 20,000-fold). The ranges of relative potencies of apical (fertilization success, spawning success, sex ratio, body length) and secondary endpoints (coloration, OSI, vitellogenin (VTG) induction) are also similar (1000 to 40,000-fold) versus 1000 to 15,000-fold). Among apical endpoints, the single lowest relative potency was 1000-fold for body length and the highest was 37,000 for fertilization success in a life cycle test with zebrafish. 

Overall the in vitro and in vivo data show that OP is considerably less potent than either E2 or EE2. These data show that the statement in the Report at Section 6.2.1.2 that OP is nearly as potent as E2 (within a factor of 2 to 28-fold) is clearly incorrect and an artifact of using unreliable studies and improper analysis of the data.   

See Table 2. Summary of Relative Potency Ratios for E2/OP and EE2/OP in attached Comments 

4.0	Examination of the estrogenic potency and aquatic toxicity of natural estrogen (E2) and synthetic estrogen (EE2) provides an understanding of what constitutes a highly potent estrogen.  

Relates to Sections 4, 5 and 6 of the Annex XV Report 

The Annex XV Report emphasizes the estrogenic activity of OP as the basis for “equivalent concern” in its assessment of the compound’s aquatic toxicity. In light of this concern, it is of use to first examine effects on aquatic organisms from known endocrine active substances for which the mode of action is indisputable – natural estrogen (E2) and a synthetic and therapeutic estrogen (EE2). Relevant studies are summarized in the table below (Table 3) and more fully presented with experimental details in the attached Table A.  E2 and EE2 have been found to be of generally similar potencies to each other. EE2/E2 ratios range from 0.19 to 1.9 depending on the screen used (Kidd et al., 2007).  E2 and EE2 are especially potent when compared to the weak estrogenic activity of OP. 

The natural and synthetic estrogens E2 and EE2, respectively, cause significant and severe effects on freshwater and saltwater fish. As described in more detail below, a signature set of aquatic effects from E2 and EE2 may include:  complete feminization of male gonads; induced failure to differentiate into males; reduced reproduction capabilities; severe histological effects on gonadal tissues; and even collapse of whole populations in a lake continuously dosed with EE2 for three years. 

A series of short term reproduction assays showed that survival, reproduction, fertilization, spawning and hatch success, and sex ratio can be significantly altered at concentrations down to 0.005 μg/L. Secondary endpoints such as gonadal weights and histopathology, testis-ova, and kidney lesions were affected at even lower concentrations, down to 0.0002 μg/L.

Life cycle studies with fathead minnow and zebrafish that included exposure to EE2 over the full life cycle showed similar effects at 0.001 to 0.016 μg /L and 0.0003 to 0.010 μg /L, respectively. Second generation NOEC were seen at concentrations as low as 0.0002 to 0.0003 μg /L (Langer et al., 2001; Wenzel et al., 2001). Secondary endpoints including secondary sex characteristics, testis-ova, gonadal histopathology, and VTG had NOEC in the 0.001 to 0.004 μg /L range. In both studies, and presumably due to the strong estrogenic potencies of these compounds essentially complete feminization of males and/or male characteristics were reported. 

In order to differentiate the relationship of endpoint effects caused by a narcotic mode of action and those mediated by an estrogenic mode of action, Wenzel et al . (2001) examined the ratio of the acute LC50 of EE2 (1,700 μg /L) and the most significant reproduction NOEC (fertilization success, NOEC 0.0003 μg /L). The acute-to-chronic ratio of the two is 5.73 million. This very high ratio is indicative of the mode of action associated with the synthetic estrogen EE2 and very different from the ratios that are calculated with OP (30 to 63) as is discussed below. 

In a whole lake study, Kidd (2007) and colleagues dosed a whole lake (0.005 to 0.006 μg/L) with EE2 for 3 years. Fathead minnow populations became completely feminized, reproduction halted, and the population collapsed. The abundance of pearl dace fish was also decreased. These two species of fish are predominant in the lake. As noted in laboratory studies, severe effects on gonadal tissues were found. Male gonadal tissue development was stunted or halted, edema was noted in ovarian tissues and lesions were found in kidney tissue. 

See Table 3 Summary of Aquatic Toxicity Studies for 17β-ethynylestradiol (EE2) and 17β-estradiol (E2) in attached comments. 

In summary, the effects on aquatic organisms by potent estrogenic substances such as E2 and EE2 collectively add up to a predictable, cohesive signature of effects, which are consistent with the known mechanism of action for these compounds. Natural development of male gonadal tissue is halted, differentiation into male characteristics is blocked, male secondary sex characteristics are prevented from developing, and gonadal tissues are severely compromised and damaged, reproduction within natural populations can be halted, and effects that are mediated by estrogenic activity occur at concentrations that are millions of times lower than those where narcotic effects occur. As a scientifically rigorous review of relevant studies shows (see Section 6.0 below), the effects of OP on aquatic organisms are very different from the effects from potent estrogens.  

5.0	The scientific evidence presented in the Annex XV Report for OP is in some cases  based on invalid studies and flawed study interpretation and the overall assessment does not adequately support a case that this compound rises to a level of concern that is equivalent to a CMR Cat.  1A, 1B, PBT or vPvB compound. 

Relates to Sections 5 and 6 of the Annex Report 

The bulk of the Annex XV Report is devoted to presenting a case that OP is a substance of “equivalent concern”; however the scientific analysis used to make that case is circumstantial, simplistic, incomplete, and in some cases wrong. Specific examples of these problems are given in the discussion of the individual studies below. A full analysis of the available aquatic toxicity data for this compound does not give rise to a conclusion that serious aquatic effects are probable or would be comparable to a Cat. 1A or 1B CMR, PBT or vPvB substance.  

5.1	There are problems with study quality scoring in the Annex XV Report on OP

In Chapter 5 of the Annex XV Report for OP, the aquatic hazard assessment is presented. The assessment introduces a number of studies mostly with fish, amphibians, and invertebrates and identifies potential findings that might be indicative of endocrine activity. Prior to analysis of any given study, the authors of the report gave each study a study quality score according to the principals of Klimisch (1997). The scoring of the various studies by the authors of the report was inappropriate in many cases. Types of errors include scoring studies as valid (scores 1 or 2) that used no statistics, lacked replication, or had poor control performance. Additionally, several studies were scored as “not valid” (3), but the studies were used anyway in the Report. Studies properly scored as “not valid” (3) cannot be used in a hazard assessment. All studies presented in the Report are captured in the attached Table B and have been re-scored following these practices.

5.2	There are problems with statistical significance and interpretation of LOEC. 

A second issue with the analysis of individual studies is the use in the Report of “Starts at”. In other words, presenting results, as positive findings, that were in fact not positive because they were not statistically different from controls. All the OECD guidance for conducting of aquatic studies, from screening studies to definitive multi-generation studies, call for adequate replication of controls and treatments such that meaningful statistical analysis can be performed. Effects are determined for any endpoint only by demonstrating with proper statistical analysis that controls and treatments are different. In addition, findings of statistical significance must be consistent and dose-related in order to establish NOEC and LOEC. For instance, statistically significant results for an endpoint at a middle dose, but not at higher or lower doses is not considered a consistent and dose or treatment related finding. The intention of the experimental designs of the OECD aquatic testing guidance when toxicity is expected is to test a series of concentrations in order to obtain a dose-response curve. The treatments are each compared using proper statistical analysis to controls and NOEC and LOEC obtained if possible. 

All studies presented in the Report are captured in the attached Table B and have been re-evaluated following these practices. The re-evaluation of individual studies includes the determination of all statistically significant endpoints that are consistent and treatment related. 

6.0	A scientifically rigorous evaluation of the “valid” or “use with care” studies on the aquatic toxicity of OP shows a consistent range of effect concentrations across species and taxonomic groups as well as across endpoints and sensitive life stages.  

Relates to Sections 5 and 6 of the Annex Report 

	6.1 	Fish

Aquatic toxicity results with OP are available for several species of fish (see the summary table below (Table 4) and attached Table B for fuller descriptions of the studies). For the screening or short term tests, specific experimental conditions varied, but adults were exposed generally from 21 to 60 days. In other tests, fertilized eggs were used to initiate the test, ensuring exposure at early and sensitive portions of the life cycle. During or following exposures numerous apical and secondary endpoints were measured. Apical endpoints included survival, growth and development, and reproduction endpoints. Many of the studies conducted reproduction trials either during or following exposure. Reproduction endpoints included egg production, spawning success, fertilization success, and hatching success. Growth and development endpoints included embryo-larval and juvenile growth, gonopodium length, and sex ratio. Secondary endpoints included those at the biochemical, plasma, or tissue level of organization. These include concentrations of vitellogenin (VTG), gonadal somatic indices, testis-ova, gonadal histopathology, and histology of other organs. 

Screening and Short Term Reproduction Studies – For these types of studies, across all apical endpoints, NOEC ranged from 10 to 1000 μg/L, while secondary endpoint NOEC ranged from 1 to 200 μg/L. By category of apical endpoint, reproduction and growth and development endpoints had similar range of NOEC (12 to 1000 and 12 to 900 μg/L, respectively), while survival NOEC ranged from 10 to 300 μg/L. Survival and secondary endpoint NOEC were similar at 10 to 300 and 1 to 200 μg/L, respectively. 

Life Cycle Tests – Life cycle tests represent the most thorough means of evaluating exposure to fish at all critical windows of their life cycle. Fully valid life cycle tests have been performed with Japanese medaka and zebrafish. 

The life cycle study with Japanese medaka was conducted by the Japanese Ministry of the Environment (Japan MOE 2002) and was not available for review. The results discussed here were presented in the Annex XV report. This study exposed newly fertilized eggs to OP throughout development to adults, followed by reproduction and early life stage exposure. The study analyzed the apical endpoints of F0 adult survival, fecundity, fertilization, hatch success, time to hatch, F0 adult growth, and F1 juvenile growth. In addition, the secondary endpoints of testis-ova and VTG induction were measured. No effects on survival, hatch success, time to hatch or F0 and F1 growth were found (highest concentration tested of 82.3 μg/L). Fecundity and fertilization were reduced at 82.3 μg/L, but not at 30.4 μg/L. Thus, the overall lowest NOEC for the study based on apical endpoints was 30.4 μg/L OP. NOEC for the secondary endpoints of testis-ova and VTG induction were ≤9.9 μg/L. Secondary endpoints such as biomarkers are generally more sensitive than apical-type endpoints, due in part to compensatory mechanisms that exist at increasing levels of biological organization (Toft and Baatrup, 2001). This was shown, for instance, in a partial life-cycle reproduction study with sheepshead minnow with EE2 (Zillioux et al., 2001) in which effects on the secondary endpoints testis-fibrosis and testis-ova were reported to occur at 2 to 20 μg/L, while effects on the apical endpoints of reproductive success and hatching success occurred only at 200 μg/L. 

The second life cycle test with zebrafish exposed newly fertilized embryos to OP through development to adult, followed by reproduction and early life stage testing. Apical endpoints of F0 and F1 early life stage survival and growth, F0 sex ratio, F0 time to spawn, F0 egg production, and F0 fertilization were measured. No effects at any concentration up to the highest tested of 35 μg/L were found on F0 embryo-larval survival, F0 larval length, F0 juvenile survival, F1 embryo-larval survival, or F1 embryo-larval length.  Reduced F0 juvenile length, F0 time to spawn, F0 egg production, F0 fertilization, and F0 sex ratio (fewer males) were reduced at 35 μg/L, but not at 12 μg/L. No secondary endpoints were recorded. These data represent scientific evidence that OP does not cause “probable serious effects” to the environment that are equivalent to CMR Cat. 1A and 1B, PBT, or vPvB substances, because the concentrations of OP required to cause adverse effects with population-level relevance in these fish species are several orders of magnitude above the concentrations of OP that are commonly measured in the environment (discussed further below).

See Table 4. Summary of Aquatic Toxicity Studies with Fish for Octylphenol (OP) in attached comments 

6.2   Amphibians

There have been several studies with amphibians and OP (summarized in attached Table C) that have been scored for reliability as (1) “valid” or (2) “use with care”. Studies scored as either (3) “not valid” or (4) “insufficient information” should not be used for hazard assessment. Explanation for reliability scoring of studies cited in the Report is given in attached Table C. 

Bevan et al. (2003) exposed embryos of African clawed frogs to OP. Effects on body length were noted at 100, but not 20 μg/L, while developmental abnormalities were reported at 1000, but not 100 μg/L. The study with the same species by Kloas et al. (1999) is considered not valid as the authors failed to control temperature during the test, had no analytical confirmation of test concentrations, had skewed sex ratio in controls, and used improper statistics. The study with the same species by Van Wyk et al. (2003) is considered not relevant as the authors employed intraperitoneal injection of OP as the means of dosing. 

Three studies used the Northern leopard frog exposing tadpoles of various stages until stage 36 when hind limb development starts. Crump et al. (2002) reported no effects at any concentration up to 200 μg/L on body weight, development, or hind limb emergence. The authors also reported a 7-d LC50 of 577 μg/L. Since the growth and developmental endpoint NOEC were ≥200 and the narcosis endpoint of lethality was 577 μg/L, the mechanism of action on the former endpoints is more likely to be narcosis related.  Croteau et al. (2009) reported only transitory increases in body weight and development at stage 29, which disappeared at stage 34. No linkage was possible of thyroid receptor activation to body weight or development. Hogan et al. (2006) reported LC50 values of 293 and 598 μg/L for stage 26 and 36 tadpoles of the Northern leopard frog. Hogan et al. (2006) also tested wood frog tadpoles using the same procedures and reported a stage 26 LC50 of 153 μg/L. As noted by the authors, possible effects on body weight cannot be discerned due to excessive crowding within test chambers, which can cause stress-related effects on growth. 

A study with the North American bullfrog by Mayer et al. (2003) exposed stage 32 to 36 tadpoles to 0.2 to 20 μg/L OP for 24 hours. Transient earlier sex differentiation occurred at stages 32 to 34, but disappeared entirely by stage 35. No effects on sex ratio from any concentration of OP were found. 

Rohr et al. (2003) exposed the streamside salamander to 5 to 500 μg/L OP for 37 days. Time to hatch, survival, and growth were reduced at 500, but not 50 μg/L OP. 
 
Overall, effects on amphibians occur in the same concentration ranges of OP as those reported with fish. From the “valid” or “use with care” studies that reported consistent and treatment related effects on amphibians, effects on tadpole development were transient and usually occurred in the same range of concentration causing lethality. Among the studies, there was no linkage of any effect to an estrogenic or thyroid-mediated causation. 

6.3	 Invertebrates
There have been several studies with invertebrates and OP (summarized in attached Table C) that have been scored for reliability as (1) “valid” or (2) “use with care”. Studies scored as either (3) “not valid” or (4) “insufficient information” cannot be used for hazard assessment and explanation for reliability scoring is also given in Table C. The aquatic toxicity of OP has been studied with Daphnia magna, copepods, sea urchins, and snails. 

Two studies with Daphnia magna are shown in attached Table C. Zou and Fingerman (1997) exposed neonates for five days at 10 to 40 μg/L and measured effects on molting. No effects on molting were  reported at any concentration. Huels AG (1992) reported the results of a 21-d life cycle study with Daphnia magna that was conducted according to OECD guideline 202. The study found effects on survival and reproduction at 100, but not 30 μg/L OP. 

With the copepod Tigeropsis japonicus, Marcial et al. (2003) conducted a two-generation test with 0.01 to 10 μg/L OP beginning with nauplii. No consistent and treatment related effects were found in F0 organisms for the endpoints nauplii stage, maturation stage, fecundity, sex ratio or survival. In F1 offspring, developmental delays (nauplii and maturation stages) were noted at most concentrations, but no effects were found on  mature copepods for fecundity, sex ratio or survival (NOEC ≥10 μg/L). 

Arslan et al. (2007) and Arslan and Parlak (2007) reported the results of studies with two sea urchin species Arbacia lizula and Paracentrosus lividus. Due to apparent solvent control effects that were not accounted for in their analysis, the Arslan et al. (2007) study with Arbacia lizula cannot be used as the study was scored as (4) insufficient information. In the study with Paracentrosus lividus (Arslan and Parlak, 2007), sea urchin sperm and eggs were exposed for 72 hours to 5 to 160 μg/L OP.  Developmental effects were found at 20 but not 10 μg/L. A 72-h EC50 for embryotoxicity was estimated from a plot to be 20 μg/L. 

A study with a third sea urchin species, Strongulocentrotus purpuratuns was reported by Roepke et al. (2005). The authors exposed sea urchin eggs to 0.0001 to 5 μg/L OP for 96 hours. An EC50 for developmental effects was reported at 0.174 μg/L. This outlier result is considered unusual and possibly suspect as the corresponding EC50 values for the natural and synthetic estrogens E2 and EE2 from the same study were 14.2 and 30.3 μg/L, respectively. In addition, the EC50 for OP was orders of magnitude more sensitive than other compounds tested by the authors including bisphenol A (EC50 226.5 μg/L). No concentrations of OP or any other of the compounds tested were analytically confirmed. Regardless of the validity of the OP EC50, this study reports results that suggest that developmental effects with this sea urchin species, Strongulocentrotus purpuratuns, are not estrogen-mediated effects. 

There are no reliable (“valid” or “use with care”) studies with mollusks that tested OP. As correctly noted in the Annex XV Report, the studies by Oehlmann et al. (2000) with giant ramshorn snails and dogwhelk and by Jobling et al. (2003) and Duft et al. (2003) with New Zealand mud snail are all properly scored for reliability as (3) “not valid”. All four of the studies suffer from numerous experimental challenges or deficiencies and so cannot be used in a hazard assessment of OP. More details are given in attached Table C. 

	6.4 	Summary of Aquatic NOEC for OP

In conclusion, there are reliable toxicity studies for fish, amphibians, and invertebrates that tested OP. The studies covered all parts of the test organisms’ life cycles from eggs to reproducing adults and cover life stages likely to be sensitive to an endocrine mode of action. Test procedures included screening tests, short-term reproduction tests, and full life-cycle tests. A consistent and treatment-related set of NOEC and LOEC have been reported and range from 10 to 900 μg/L across the apical endpoints related to survival, growth and development, and reproduction. 

7.0 	The Report does not establish that the reproductive and other aquatic toxicity effects of 4-t-OP are solely - or primarily - caused by an endocrine mode of action and an evaluation of the aquatic toxicity of OP does not support concern for “probable serious effects” to the environment that are equivalent to CMR Cat. 1A and 1B, PBT, or vPvB substances.

Relates to Sections  5 and 6 of the Annex Report and to the Report in general

The Report concedes that the question of how to define “probable serious effects to the environment” and how to conclude whether or not they are “of equivalent concern” is still the subject of debate.  While the Report does not suggest any general criteria for these terms, it suggests that the criteria in this case should take into account the potency of the substance and the relevance of the observed effects.  As such the Report develops the case in the OP Annex XV Report around two questions:  
1. Is OP a substance having endocrine disrupting properties?
2. Is there scientific evidence of probable serious effects which give arise to an equivalent concern compared to CMR and/or PBT substances?

Based on the language in Art. 57 it is clear that “equivalent concern” should not rise solely from the mode of action (i.e. endocrine). Rather the effect (i.e. apical endpoint or a population level effect) must be of “equivalent concern” to those of CMR Cat. 1A, 1B, PBT or vPvB compounds.   The Annex XV Report for OP incorrectly suggests that the effects of OP are equivalent to those of known potent estrogens, provides inadequate rationale for a linkage between the very weak estrogenic activity of OP to reproductive or developmental effects in aquatic organisms, and incorrectly attributes obvious general toxicity in amphibians to thyroidal activity. The following comments address these points. 

7.1	OP affects acute lethality and chronic reproduction, growth and development endpoints very differently than natural (E2) or synthetic (EE2) estrogens.

Considerable effort has been made in the Annex XV Report to link effects reported for OP from all the studies with fish to an endocrine disruption mode of action.  Comparison of studies examining OP and natural (E2) and synthetic (EE2) estrogens demonstrates that there are great differences between the effects that they cause to aquatic organisms.

In a life cycle test with zebrafish, Wenzel et al. (2001) demonstrated that estrogenic activity for the natural and synthetic estrogens E2 and EE2 occurs at concentrations that are orders of magnitude below the threshold for systemic toxicity, and result in a suite of responses that are collectively linked to the estrogenic mode of action. In the life cycle study with EE2 and zebrafish, Wenzel et al. (2001) reported the 96-h LC50 (narcosis-based toxicity) for EE2 to be 1700 μg/L and the NOEC for fertilization success to be 0.0003 μg/L. The authors calculated the ratio of the two values to be 5.73 x 106. It is clear for EE2 that the reproduction, growth and development endpoints are affected by a different mechanism of action (endocrine) than the acute lethality (narcosis) endpoints. Given the potency of natural and synthetic estrogens, this is expected, and hence the very high ratio is understandable. 

OP does not affect the same endpoints in aquatic organisms in the same way that natural and synthetic estrogens act. Data from three studies with OP demonstrate that the reproduction, growth and development endpoints are mainly indicative of narcosis-type mode of action, possibly coupled with a very weakly estrogenic mode of action. 

In the life cycle study with OP and zebrafish, Wenzel et al. (2001) reported a 96-h LC50 of 370 μg/L. The ratio of the acute LC50 and the lowest apical development NOEC from this study (fertilization success) of 12 μg/L is 31. In the life cycle study with OP and Japanese medaka, the Japan MOE (2002) did not report an acute value. However, while an LC50 for Japanese medaka is not available for OP, it can be assumed to be about 900 μg/L, based on a review of many data for OP and the structurally similar nonylphenol (NP) that ranged up to 900 μg/L across many freshwater and marine species (Staples et al., 1998). Using the data from the Japanese MOE (2002) report, the ratio of an estimated LC50 of 900 μg/L and the NOEC of fertilization success (30.4 μg/L) was calculated to be 30. For sheepshead minnow, Karels et al. (2003) reported a 72-h LC50 of 720 μg/L for OP, while the NOEC for effects on percent viable eggs was 11.5 μg/L OP. The ratio of the two values is 63. Testing guppy, Toft and Baatrup (2003) reported a 96-h LC50 value of 495 μg/L for OP. A ratio of 50 was calculated using the LC50 and the NOEC of 10 μg/L for the most sensitive reproductive or developmental endpoint, gonopodium length. Thus, the ratio of acute lethality LC50 values and the NOEC of the lowest apical NOEC for reproduction, growth or development endpoints are consistent across the species zebrafish, Japanese medaka, sheepshead minnow, and guppy. Across the four species, the ratios range from 30 to 63, which are approximately 100,000- to 200,000-fold lower than the ratio calculated for EE2. 

Together these data show that OP has a very different mode of action than the natural and synthetic estrogens with effects on survival resulting from narcosis being at least as critical to the organisms exposed to OP as other modes of toxicity. While biomarkers such as VTG indicate that estrogen receptor binding is occurring while exposed to OP, the endpoints related to growth, development, and reproduction are only a factor of 30 to 63 from short term acute toxicity and in the same range as caused by longer term lethality.  Hence it is possible that effects on growth, development, and reproduction for OP are more indicative of narcosis-type mode of action, possibly coupled with a very weakly estrogenic mode of action. Taken together, the acute and chronic datasets for OP and EE2 are very different. 

7.2	The concern expressed in the Annex XV Report on OP is for reproductive or developmental effects in aquatic organisms caused by an endocrine mechanism; however it does not provide adequate rationale for the leap between findings of weak 4-t-OP estrogenic activity and reproductive effects seen in some fish studies. 

The endpoints that are considered to indicate an estrogen agonist mode of action are summarized on page 104 (Table 27) of the Report.  Several of these endpoints are more apical in nature and can be observed naturally and also under various stressful conditions for fish.  Oocyte atresia normally occurs in fish as a part of the natural reproductive cycle, so an understanding of the normal background level of this phenomenon is important. Oocyte atresia can also occur in the ovaries of female fish when exposed not only to an estrogenic compound, but also in response to an androgenic mode of action (Ankley et al., 2003).  Therefore it appears that oocyte atresia, in and of itself, is not indicative of a particular endocrine mode of action.  In addition, a depression of secondary sex characteristics in the fathead minnow or medaka has also been observed in toxicity tests with compounds that operate via a mode of action that is not related specifically to estrogen agonism (USEPA, 2007).  
In various studies with medaka fish, increased VTG was observed in males at concentrations of OP ≥ 11.4 µg/L, altered histopathology of the gonads (specifically occurrence of testis-ova) generally occurred at concentrations of OP ≥ 11.4 µg/L (Gronen et al., 1999; Seki et al., 2003; Gray et al., 1999a; Gray et al., 1999b).  The occurrence of testis-ova in the gonads was more marked when fish were exposed during the time period of sexual differentiation, which occurs before hatch.  Incidences of ova-testis were less frequent and occurred at higher concentrations when exposure to OP was initiated post-hatch (Gray et al, 1999a; Seki et al., 2003).  Male fish with testis-ova were observably undergoing active spermatogenesis and were able to fertilize a female fish, so it appears that the occurrence of testis-ova does not preclude reproduction (Gray et al., 1999b; Seki et al., 2003).  Growth and reproduction were adversely affected at concentrations of OP ≥ 20 µg/L (Gronen et al., 1999; Seki et al., 2003; Gray et al., 1999a; Gray et al., 1999b).  Sex ratios in medaka fish were altered toward a greater proportion of females at concentrations of OP ≥ 48.1 µg/L (Seki et al., 2003; Gray et al., 2009).  As noted by Gray et al. (1999a), effects on apical endpoints that would indicate adverse effects on a population level parameter (such as altered sex ratios, decreased growth and reproduction) occurred at concentrations of OP that are higher than levels found in the environment.  
In guppies exposed for 90 days to nominal concentrations of 1 to 200 μg/L of OP, the lowest concentration to cause statistically significant effects (increased gonopodium length, increased sperm count, decreased ovary weight) was 100 μg/L (Toft and Baatrup, 2003).  This concentration, however, resulted in up to 40% mortality.  The authors stated this concentration of OP was considerably higher than what is found in the environment and concluded “exposure of guppies to environmentally realistic concentrations of OP does not alter the sexual characteristics of these fish”.  When male and female guppies were exposed to 26 μg/L OP for 28 days (males) and 26 to 36 days (females), no differences were observed with survival, gonadosomatic indices, brood intervals, brood size, gonopodium length and sex ratio of the offspring even though there were potential changes to the testis structure (indications of blocked spermatogonial mitosis) in the males (Kinnberg et al, 2003).
In regards to the amphibian data set, the Kloas et al. (1999) study was given a reliability score of (3) “not valid” for several reasons as described in Section 6.2 of these comments. In addition, the Kloas et al. (1999) study should not be considered valid for use with respect to the altered sex ratio endpoint, because gonads of juvenile Xenopus laevis were only inspected based on gross morphology and not on histopathology.  Histopathological examination of the gonads is needed to correctly verify sex, especially in juvenile organisms such as those examined in this study. In addition, due to the non-relevant nature of the administered dose (via injection) and the small sample size, the van Wyk et al. (2003) study is of limited relevance.   In the studies with amphibians that were considered to be valid for use, there were indications of population level effects (on endpoints such as growth, reproduction, survival) at concentrations of OP < 50 µg/L (stage 36).  However, this effect at 50 µg/L (increased body weight) was noted in one stage of Rana pipiens tadpole development and was not noted in other developmental stages with the same species and was not noted among a closely related frog species (e.g., R. sylvatica) in the same experiment (Hogan et al., 2006).  More generally, adverse effects in amphibians were apparent at concentrations of OP ≥ 100 µg/L (Bevan et al., 2003; Rohr et al., 2003), levels that are three orders of magnitude greater than the upper 90th centile concentration of 0.25 µg/L OP in European surface waters.  

In a more recent, well-conducted, chronic study with OP and Xenopus tropicalis, frogs were continuously exposed to 0,  1.1, 3.3, 11, and 36 µg/L OP from Nieuwkoop and Faber stage 46 to adulthood (a total of 31 weeks) with a sampling time-point at the completion of metamorphosis as well (Porter et al., 2011).  In this study, no significant deviation from the control sex ratio was observed for either sampling period, suggesting minimal to no effect of OP exposure on gonad differentiation.  No effects in the adult frogs were observed for mortality, body mass and size, liver somatic index, estradiol and testosterone serum levels, sperm counts, or oocyte counts. The development and growth of oviducts, was observed in males exposed to OP, and VTG levels were increased among juvenile male frogs (sampled at the time of metamorphic completion), but not in adult male frogs at the close of the exposure at 31 weeks.  The transient increase in male vitellogenin and the presence of oviducts in some male frogs exposed to OP indicates weak estrogenic activity of OP, however Porter et al (2011) concludes that the highest OP concentration used, 36 µg/L, was at or below the no observed effect level (NOEL) for general toxicity in X. tropicalis.  

Together, the studies with fish and amphibians did not fully link secondary endpoints such as testis-ova or VTG induction to adverse outcomes. Fish with testis-ova were able to reproduce and male frogs with increased VTG levels developed normally. The Report does not provide adequate rationale for fully connecting findings of weak 4-t-OP estrogenic activity and reproductive or developmental effects seen in some fish studies.

7.3    The case made in the Annex XV Report for OP exhibiting thyroidal activity in
         amphibians is weak and inconclusive, and is not sufficient to warrant OP as being
         of “equivalent concern” as CMR and PBT or vPvB.  

The Annex XV Report states, “4-tert-octylphenol alters the function of the endocrine system in aquatic organisms through an estrogen mediated mode of action, with some evidence of thyroidal activity in amphibians” (pg 6).  However, as will be discussed, the case for OP exhibiting thyroidal activity in amphibians is weak and inconclusive, and is not sufficient to warrant OP as being of “equivalent concern” as CMR and PBT or vPvB.  

The Annex XV Report correctly indicates that the endpoints in the 21-day amphibian metamorphosis assay (OECD Guideline 231) that are intended to indicate potential thyroid activity are: advanced development (measured by advanced developmental stage or increased hind limb length relative to controls), asynchronous development, and/or significant histopathologic alterations of the thyroid glands.  Out of the eight amphibian studies summarized in the Annex XV Report, only studies on two frog species are referenced on page 6 of the Report as providing some evidence for a thyroid agonist mode of action for OP.  These studies include testing on Rana pipens and Rana catesbeiana tadpoles.   These studies will be closely examined for the strength of an OP association with thyroid agonism/potential thyroid activity.

In one study, Rana pipiens tadpoles exposed to OP for eight months showed delayed development and higher body weights (at stage 29 only) starting at 0.02 µg/L OP, increased levels of thyroid receptor β mRNA (at stage 34, but not 29 or 42) at concentrations of 2 µg/L OP, and increased deiodinase III mRNA (at stage 29, but not 34 or 42) at 0.02 but not 2 µg/L OP (Croteau et al., 2009).  Thyroid receptor β binds thyroid hormones and is directly autoinduced by thyroid hormone binding (Denver, 1998; Hogan et al., 2007; Zhang et al 2006).  Deiodinase III inactivates thyroid hormone (Denver, 1998; Hogan et al., 2007).  There were no significant alterations in thyroid hormone levels among stage 29 or 34 tadpoles in this study.  Taken together, the occurrence of increased deiodinase III mRNA at stage 29 (at the low dose of OP), the increased thyroid receptor β mRNA at stage 34 only, the lack of alteration in circulating levels of thyroid hormone levels, and delayed development do not indicate a consistent or cohesive picture of perturbation of thyroid activity in association with OP exposure in the Croteau et al. (2009) study.  Specifically, increased thyroid receptor β mRNA should correspond with increased thyroid hormone levels and advanced, not delayed, development..  Another consideration for this particular study with R. pipens is that the tadpoles were reared at relatively high densities, which may have resulted in stress to the tadpoles potentially interfering with measured endpoints (e.g., increased rearing density increases corticosterone (Glennmier and Denver 2002a), which can slow growth and development (Glennmier and Denver, 2002b).  Also, no positive control was used in this study design to establish what responses would be expected for a known thyroid active compound in this species with the selected endpoints.  In another study with R. pipens tadpoles, shortened time to hind limb emergence (advanced development) was observed among tadpoles exposed to 200 µg/L OP and ultraviolet B (UVB) radiation for 10 days.  This same effect was not noted when tadpoles were only exposed to OP (and not UVB) for 10 days (Crump et al., 2002).  Therefore, there was no indication of thyroid agonism for OP acting alone, apart from UVB radiation (Crump et al., 2002).    

 In another study, Rana catesbiena tadpoles were exposed to OP for 24 hours and an earlier completion of sexual differentiation was observed at concentrations of OP ≤ 0.2 µg/L (Mayer et al., 2003).  However, there is no definitive link between an earlier completion of sexual differentiation and thyroid agonism and/or thyroid perturbation.  The process of gonad differentiation in amphibians appears to be controlled separately from activity in the hypothalamus-pituitary-thyroid axis (Robertson and Kelley, 1996; Buchholz et al., 2006; Coady et al., 2010).  Therefore, this study should not be used to support the potential thyroid agonsim of OP.  Furthermore, in a recent well-designed chronic study wherein the western clawed frog (Xenopus tropicalis) was exposed to four concentrations of OP (ranging from 1 to 36 µg/L) beginning at premetamorphic developmental stages and lasting until sexual maturity (31 weeks), there were no effects on gonad differentiation in OP exposed individuals relative to controls (Porter et al, 2011).  Thus, there are no definitive data to indicate that OP acts on the thyroid system of amphibians either as a thyroid agonist or antagonist, and this line of evidence should not be used to indicate that OP has endocrine disrupting characteristics that are of equivalent concern to CMR, PBT or vPvB classification status.

In summation, results of aquatic toxicity tests conducted OP and EE2 show that: 
1) effects on acute lethality, reproduction, growth, and development are very different for the two substances; 
2) studies with fish and frogs exposed to OP failed to fully link an endocrine mode of action affecting secondary endpoints with adverse reproductive, growth or developmental outcomes; and 
3) there are no definitive data to indicate that OP acts on the thyroid system of amphibians either as a thyroid agonist or antagonist. Together, these data represent further scientific evidence that OP does not cause “probable serious effects” to the environment that are of equivalent concern to CMR Cat. 1a and 1b, PBT, or vPvB substances.

8.0   	The arguments listed in the summary of the Annex XV Report on OP are not supported by an adequate scientific basis to propose identifying this compound as a Substance of Very High Concern (SVHC) on the basis of “equivalent concern” for endocrine modulated environmental effects.

Relates to the Summary and Section 6 of the Annex Report and to the Report in general

The Annex XV Report states that OP “is proposed to be identified as a “substance of equivalent concern according to Article 57 (f) because of its environmental endocrine disrupting properties and subsequent serious effects to the environment” and lists a series of arguments or concern as a basis for this proposal. However, these arguments are not supported when considered relative to the data provided previously in these comments. The following comments respond to the arguments listed as the basis for this proposal.   

8.1	 The case made in the Annex XV Report for OP exhibiting thyroidal activity in amphibians is weak and inconclusive, and is not sufficient to warrant OP as being of “equivalent concern” as CMR and PBT or vPvB.  
The Report summary states: 
•	tert-octylphenol alters the function of the endocrine system in aquatic organisms through an estrogen mediated mode of action, with some evidence of thyroidal activity in amphibians”  
•	 Effects that occurred in two other amphibian species (R. catesbeiana and R .pipens) provide some evidence for a thyroid agonist mode of action. Both effect types indicate that 4-tert-octylphenol might be an endocrine disruptor in amphibians
As discussed in Section 7.3 of these comments, the case made in the Annex XV Report for OP exhibiting thyroidal activity in amphibians is unsubstantiated. Therefore, there is no basis to support the above summary statement and the data are not sufficient to warrant OP as being of “equivalent concern” as CMR and PBT or vPvB.  

8.2 	Adverse effects in fish species are not “clearly endocrine mediated”; rather they are more indicative of general toxicity possibly coupled with very weakly estrogenic activity. 

The Report summary states: 

•	Adverse effects observed in three fish species are clearly endocrine mediated
•	In all other tested fish species the endpoints affected are known to be influenced by estrogen activity and, with only one exemption, an estrogen mediated mode of action was observed.
•	Ranges of NOEC were essentially the same for survival, growth and development, and reproduction endpoints

The first summary statement listed above refers to studies with Japanese medaka, sheepshead minnow, and guppy fish. The following table (Table 5) shows that more than half of the NOEC for these species were actually "no effects at any concentration of OP tested". The range of concentrations showing effects in fish was about 20 to 900 µg/L OP.

See TABLE 5  Summary of effects from exposure to octylphenol for three fish species in attached comments 

As discussed in Section 5.0 of these comments, there were issues with study validity scoring and interpretation in the assessment of fish data in the Annex XV Report.  The endpoints showing effects (survival, growth, and reproductive endpoints) are not necessarily "endocrine mediated". Interpreted properly, the data show that the ratio of acute lethality (LC50) to the lowest reproductive NOEC for OP were 30 to 63 for medaka, sheepshead minnow, and guppy. A similar ratio of 31 for zebrafish is also reported. The comparable ratio for the potent synthetic estrogen EE2 with zebrafish is 5,700,000. The vast difference in these ratios show that effects caused by OP are not "clearly endocrine mediated", but rather are indicative of general toxicity possibly coupled with very weakly estrogenic activity. 

8.3   Effects observed for studies of OP in amphibian species are not comparable to
        those observed for 17ß-estradiol.
     
The Report summary states: 
•	the effects observed in the amphibian species X.laevis after exposure to 4-tert-octylphenol are comparable to effects observed for 17ß-estradiol 

 As discussed in Section 6.3 of these comments, two of the three studies cited with this species were scored (3) "not valid" and are not reliable (see previous comments and attached Table B). Kloas et al. (1999) was found to have numerous experimental design flaws. Van Wyk et al. (2003) utilized intraperitonial injection of the frogs. The third study only reported effects with OP with effects on growth and development at 100 and 1000 µg/L OP, respectively; however there was no positive control and therefore no basis for comparison to E2. Therefore, there is no basis to support the summary statement above that effects observed for OP in amphibian species are comparable to E2. 
8.4	The Annex XV Report does not adequately support its argument that OP is a potent endocrine disruptor 
The Report summary states: 
•	4-tert-octylphenol is a potent endocrine disruptor with regard to the concentration required to cause endocrine mediated effects and the exposure duration needed to cause effects  
•	Tests with some fish species testing 17ß-estradiol and 4-tert-octylphenol indicated that 4-tert-octylphenol may be nearly as potent as the natural estrogen 17ß-estradiol (factor 2- 28 less potent)

As discussed in Section 3.0 of these comments, the Report does not adequately support its argument that OP is a potent endocrine disruptor. The summary statements listed above are simply not supported by the available data.   The relative potencies for OP relative to E2 or EE2 cited in the Annex XV Report (2 to 28) were not calculated correctly.  More than half of the fish and amphibian NOEC were actually no effects at any concentration of OP tested. The range of concentrations showing effects was about 20 to 900 µg/L OP. The endpoints showing effects included survival, growth, and reproductive endpoints and NOEC for these end points are all similar.  When properly calculated from reliable studies, the estrogenic potency of OP is in the range of 1000 to 40,000 less potent than E2 or EE2 based on numerous in vitro and in vivo studies. 

Ratios of acute lethality to lowest reproductive effect NOEC (acute-to-chronic) are 30 to 63 for OP, as compared to 5,700,000 for EE2. OP effects are more likely caused by general toxicity possibly coupled with very weak estrogenic activity. Studies with fish have been conducted with exposure regimes that cover all times of the life cycle from egg to embryo to juvenile to reproducing adult. All critical windows of exposure during any stage of fish development have been tested for effects from OP. 

8.5	The Annex XV Report does not adequately support its argument that OP has effects like a potent estrogen 

The Report summary states: 
•	Taking our knowledge on potent estrogen agonists into account, the level of concern for 4-tertoctylphenol seems to be comparable to PBT /vPvB and CMR substances. 
•	Estrogens are known to result in irreversible, persistent effects.

Section 4.0 of these comments examines the estrogenic potency, efficacy and aquatic toxicity of natural estrogen (E2) and synthetic estrogen (EE2) and provides an understanding of what constitutes a highly potent estrogen.  The effects on aquatic organisms by potent estrogenic substances such as E2 and EE2 collectively add up to a predictable, cohesive signature of effects, which are consistent with the known mechanism of action for these compounds. Natural development of male gonadal tissue is halted, differentiation into male characteristics is blocked, male secondary sex characteristics are prevented from developing, and gonadal tissues are severely compromised and damaged, reproduction within natural populations can be halted, and effects that are mediated by estrogenic activity occur at concentrations that are millions of times lower than those where narcotic effects occur.  In Section 6.0 of these comments a scientifically rigorous review of relevant studies shows the effects of OP on aquatic organisms are very different from the effects from potent estrogens and are not comparable to PBT/vPvB or CMR Cat. 1A or 1B compounds. 

8.6	Sufficient studies covering all critical windows of exposure during all stages of fish development have been considered in the aquatic studies for this compound.

The Report summary states: 

•	A change in the endocrine feedback system during sensitive life stages may result 
             in effects during the entire life. 
•	Impairment of reproduction may already occur after a transient short term exposure of adults as observed in two fish species (21d exposure of adult males)

As discussed in Section 6.0 of these comments, fish have been tested with OP over all critical windows of exposure and life stages likely to be sensitive to endocrine modes of action with a NOEC range from about 10 to 900 µg/L OP. The duration of these studies ranged from short term exposures to life-cycle exposures and the NOEC and LOEC values that have been developed account for any life stage sensitive effects. 

8.7	OP does not meet the bioaccumulative criteria for SVHC compounds and its “distribution toward embryos in viviparous species” did not cause developmental effects in offspring or raise concern for the maternal transfer of OP within female guppy fish.

The Report summary states “with respect to 4-tert-octylphenol the level of concern is increased by its bioaccumulation potential with some evidence of internal distribution toward embryos in viviparous species.

This supporting statement is misleading and inconsistent with the conclusion in section 3.3 of the Annex XV Report that OP is only low to moderately bioaccumulative (as evidenced by laboratory BCF results ranging from 46 to 471) and does not meet the bioaccumulation criteria to be considered a SVHC. The observation "internal distribution toward embryos in viviparous species” was not attributed to a specific study in the Report summary, but may relate to a study by Kinnberg et al. (2003) in which the authors did not find any "level of concern" related to maternal transfer of OP within female guppy fish.  The authors exposed female guppy (a viviparous species) to 26 μg/L OP or 0.85 μg/L E2. According to the authors, "At the tested concentrations, exposure to octylphenol or 17B-estradiol via the mother fish did not significantly affect the weight, length, gonopodium index or sex differentiation of the offspring". In addition, a study with the viviparous species eelpout (Rasmussen et al. (2002) examined the offspring from a female fish exposed to 0, 25, or 100 μg/L OP. Significant effects on the offspring were only found in offspring from the females exposed to 100, but not 25 μg/L OP, including sexual differentiation and VTG induction. These results are fully in line with the range of NOEC and LOEC across numerous species, do not show any unexpected results and do not warrant equivalent concern to a CMR Cat. 1A or 1B or a PBT/vPvB compound.  

8.8 	OP does not meet persistence criteria for SVHC compounds. 

The Report summary states that OP “is persistent under anaerobic conditions combined with a tendency to adsorb to sediment and soil”.   

This supporting statement is misleading and inconsistent with the conclusion in section 3.1 of the Annex XV Report. OP does not meet any of the persistence criteria for SVHC compounds. As is true for many organic substances, OP degrades more slowly under anoxic conditions. The moderate sorption capacity to soil and sediment acts to keep OP from being transported further after release into those compartments. 

8.9	Considering the low estrogenic potency of OP, along with its low occurrence and concentrations in the environment, it is not likely, or probable, that this compound will result in effects that are adverse to that ecosystem with respect to community structure and function. 
  
The Report summary states:
•	Population relevant endpoints affected in these species are known to be sensitive toward estrogen activity, although not being diagnostic for this mode of action  
•	Exposure to 4-tert-octylphenol may result in effects that relevantly influence ecosystems with respect to the community structure and function
•	Comparable to other estrogens, 4-tert-octylphenol influences reproduction parameters as well as sexual development (including changes in sex-ratio) and growth and thus endpoints are affected that may impair population stability and recruitment.
•	Several species belonging to different taxonomic groups were affected at low concentrations indicating that 4-tert-octylphenol may cause endocrine mediated effects in a variety of species.
•	Some evidence for potent estrogen agonists is available that population stability is affected even after exposure has ceased. 
•	Due to a highly conserved receptor among vertebrate species it is likely that several fish species representing a variety of reproduction strategies and migration patterns may be affected.
•	The type of effects observed (e.g., short exposure to 4-tert-octylphenol in adults affects fertilization success) provide some evidence that exposure in one area might influence population stability in another area (e.g., for salmonids migrating long distances passing highly polluted areas during migration towards spawning grounds). 

The data provided in these comments shows that OP has only weakly estrogenic activity, probably combined with general toxicity (e.g, narcosis) and is clearly not "comparable to other estrogens".  As noted above, the potency for OP relative to E2 or EE2, when properly calculated from reliable studies, is in the range of 1000 to 40,000 fold less potent across numerous in vitro and in vivo studies.  Adverse apical effects caused by OP are not "clearly endocrine mediated" but rather are indicative of general toxicity possible coupled with very weakly estrogenic activity.   All valid NOEC based on apical endpoints ranged from about 10 to 900 µg/L. Concentrations measured in Europe were reported to be 0.05 μg/L (median concentration) to 0.25 μg/L(upper 90th percentile concentration), showing that concentrations were well below the full range of NOEC for OP so the probability of OP causing population level effects is low. 	See response to comment 20	
			See also:
Comment 19 attachment (CEPAD-APERC Comments on OP SVHC (Oct. 13, 2011).zip).zip		
18	2011/10/13	National NGO
Danish Ecological Council
Denmark	We do not agree in the conclusion “4-tert-octylphenol does not represent a substance with ED properties of strong potency for the mammalian system” (page 155).
In the dossier it is indeed acknowledged that many studies indicate that 4-tert-octylphenol is an endocrine disrupter, and it is concluded “that 4-tert-octylphenol features some inherent potential for being toxic for reproduction, probably in relation to female sexual maturation and female fertility” (page 153). So with the available evidence we emphasis the importance of a clear statement that 4-tert-octylphenol should be measured as having ED properties, also for humans.	See response to COM 24 in General Comments	
17	2011/10/13	National NGO
Réseau Environnement Santé
France	Page 155] Final conclusion stating that Octylphenol is not a "substance with ED properties of strong potency for the mammalian system". 

It is acknowledged in the dossier that there are several studies that indicate that octylphenol is an endocrine disruptor in mammals with "inherent potential for being toxic for reproduction" [page 153]. Taking into consideration the evidence pointing at octylphenol as an endocrine disruptor, we therefore believe it should be clearly stated that Octylphenol with the available evidence is to be considered an endocrine disrupting chemical also for mammals, including humans.

Our uploaded annex provides additional studies supporting the endocrine disrupting properties of octylphenol in mammals. See references 1 to 6.
	See response to COM 24 in General Comments

As stated in the report, references in which the studied substance could clearly be identified as 4-tert-octylphenol e.g. by name, CAS number or structural formula were included in the assessment. Studies with ‚octylphenol’ without further specification were excluded due to unclear substance identity. Studies with 4-n-octylphenol as investigated substance were excluded.
Sainath et al. 2011: 4-tert-OP; published after end of literature search. Will be considered for future discussions. 
Turan et al. 2008: unclear substance identity (‘octylphenol’ without further specification)
Bonefeld-Jorgensen et al. 2007: substance identity: 4-n-octylphenol (as identified by name and structure)
Christian & Gillies 1999: unclear substance identity (‘octylphenol’ without further specification)
Hernandez-Rodriguez et al. 2007: substance identity: 4-n-octylphenol (as identified by CAS No)
Sweeney et al, 2000: unclear substance identity (‘octylphenol’ without further specification)	
			See also:
Comment 17 attachment (Annex RES octylphenol.pdf).pdf		
16	2011/10/13	Member State
United Kingdom	Section 3.1.3 (Summary and discussion of persistence) (p. 14): We believe that the screening tests show that there is potential for extensive mineralisation (62.1% in 28 days), but agree that this appears to require adaptation. As noted in the discussion on simulation tests, dissipation half-lives were lower in urban areas than in rural locations. In the ESR assessment of nonylphenol, it was noted that the ubiquitous presence of alkylphenols in industrial areas meant that microbes in all compartments might be expected to be acclimated to some degree. Therefore, whilst we agree that the substance may biodegrade slowly under some conditions, the conclusion in the dossier that this is appropriate for the ‘whole environment’ may be misleading. In our risk evaluation report, we concluded that, in general terms, 4-tert-octylphenol is considered to be inherently biodegradable (fulfilling the criteria), whereas it is inherently biodegradable (not fulfilling the criteria) for the purposes of the assessment of WWTP. This assumption underpins some of the exposure estimates that are reported later in the dossier.

Section 5.1.2.3 (Fishes) (p. 102-): In general, we appreciate the efforts that the German c.a. has undertaken to elucidate key endocrine effects. We know that testis-ova may sometimes appear in male control fish, and historical control sex ratios also need to be taken into account. We therefore think that some of the statements should be accompanied by the term “outside the historical control range” (e.g. ‘presence of testis-ova in male fish outside the historical control range is diagnostic for endocrine activity’). 

Table 28: It is not clear why a study that has ‘insufficient documentation’ should be considered reliable with Klimisch score 2. Were any attempts made to clarify the findings with the study authors?

Table 40: It would be helpful to indicate what effects were observed at the “effect concentration” for the two species listed.

Overall, we agree with the conclusions drawn in this section, i.e. that the substance produces adverse effects in some fish species that appear to be correlated with disruption of the endocrine system.

Section 5.1.2.4 (amphibians) (p. 132 -): The description of the study by Kloas et al. (1999) fails to mention that it was a method development study, and that a repeat experiment by another lab failed to replicate the findings for bisphenol-A, which was tested using the same system. To our mind, this casts considerable doubt on the findings. There seems to be no information on historical control variability in sex ratio either. Therefore, it is perhaps less reliable than suggested in the dossier.

Given the absence of studies performed according to ring-tested guidelines, we think the overall conclusion for amphibians is too strong. We agree that there are some indications of possible effects, but the mechanism is not clear.

Section 5.1.2.5 (invertebrates) (p. 137-): As we have remarked in our general comments, we believe the statements about the effects on molluscs are incorrect. None of the available studies is considered reliable. It is therefore misleading to suggest that there are effects mediated by any endocrine mechanism. The same comment applies to the other invertebrate results. Whilst it is clear that the substance affects reproduction in several species, it cannot be concluded that they are ‘possibly mediated by an ED mode of action’. We find the concluding remarks too speculative.

Section 6.2.1.1 (Endocrine disrupting properties) (p. 143-): We support the conclusion that 4-tert-octylphenol is an endocrine disrupter in fish. We do not agree with the statements made about amphibians or invertebrates, for the reasons given above.

Section 6.2.1.2 (Equivalence of concern) (p. 146-): We would like to thank the German c.a. for taking into account some of the issues we have raised in our developing ideas for an EDC definition. In terms of potency, we believe that the discussion should only focus on fish, because this is the only taxonomic group with sufficiently robust data. We would like to see a more systematic comparison with 17β-oestradiol, since the cited potency factor (2-28) only applies in some cases, and this should really be discussed in more detail. However, we agree that the endocrine-related effects in fish do appear to be amongst the more sensitive end points in the available data set (a summary table and/or species sensitivity distribution would help to illustrate this point more clearly). NOECs occur below 0.1 mg/l, and they appear to be within one to two orders of magnitude of other reliable NOECs that have not been identified as endocrine mediated (e.g. invertebrates).

We think the demonstration of ED effects in more than one fish species is sufficient to flag a concern for the taxonomic group in general, and we do not think the reference to amphibians or molluscs is needed. Overall, we can accept the conclusion of equivalent concern.

Our final remark concerns what would happen next, if it were agreed that 4-tert-octylphenol should be identified as an endocrine disrupter of equivalent concern. Either restriction or authorisation will need to establish a PNEC, since the effects are not non-threshold in nature. Has the German c.a. given any thought to what this PNEC might be? 

Editorial remarks

Section 2 (p. 10): The headings seemed to have been mixed up for Sections 2.1 and 2.2.

Section 3.1.2.3 (Biodegradation simulation tests) (p. 13): Terms such as DisDT50 should be explained the first time they are used.

Section 3.2.1 Adsorption/desorption (p. 14): A log Kow of 4.12 is used in this section, but this is not mentioned in Section 1. Similarly, there is a statement that the QSAR has limited applicability for surface active substances, but there is no previous discussion of the surface active properties of this substance. The measured Koc data make this opening paragraph unnecessary, but it should be amended.

We note that this Kow value is also mentioned in Section 3.3.1.1 (bioaccumulation estimation) (p. 16).

Section 3.2.3 (Distribution modelling) (p. 15): The concluding remark that “both models show that the substance will primarily partition to soil” is only true for exclusive release to air and soil. Releases to water will mainly remain in water.
	Section 3.1.3: We appreciate your comment, however, if analysed in detail, the screening tests show contradictory results (between 0% and 62.1% in 28 days), which suggests that 4-tert-octylphenol is not readily biodegradable but has a potential of biodegradation after a period of adaption. The simulation tests show no degradation but rather dissipation. The dissipation is caused by the strong adsorption of 4-tert-octylphenol to soil, sludge and sediment (under anaerobic conditions no elimination was observed in sediment). Therefore we do not agree that 4-tert-octylphenol is inherently biodegradable.

Section 5.1.2.3: Thank you for your comment. However, as common for other endpoints and described in the OECD test guidline 234 for sex-ratio, we suggest, that effects should be compared to controls not to historical control ranges. 

Table 28: With regard to Knörr and Braunbeck, 2002 some documentation was missing but enough information was provided to score a reliability of 2. With regard to the study by the ministry of Japan, 2002 the score was changed to 4 until further information is available from Japan.

Table 40: See header of the table (in vivo vitellogenin induction)

Thank you for the overall support.

Section 5.1.2.4: Thank you for your comments. The overall conclusion in sections 5.1.2.4 was rephrased in order to state more clearly that there are only some indications of a possible endocrine mode of action. 

Section 5.1.2.5: The section was revised in order to state clearly, that studies are of low quality and provide some hints only. 

Section 6.2.1.1.Thank you for your support. See comment above with respect to amphibians or invertebrates.

Section 6.2.1.2: A new paragraph was included in chapter 6.2.1.2 focusing on the relative potency for fish. A more in depth analysis was not included as the overall weight of evidence decision is not based on the relative potency

Thank you for your comment with regard to PNEC derivation. No PNEC was derived yet as this was considered not to be in the scope of the dossier.

Editorial remarks were revised accordingly. 

14	2011/10/13	International NGO
ChemSec
Sweden	[Page 155] Final conclusion stating that Octylphenol is not a "substance with ED properties of strong potency for the mammalian system". 
It is acknowledged in the dossier that there are several studies that indicate that octylphenol is an endocrine disruptor in mammals with "inherent potential for being toxic for reproduction" [page 153]. Taking into consideration the evidence pointing at octylphenol as an endocrine disruptor, we therefore believe it should be clearly stated that Octylphenol with the available evidence is to be considered an endocrine disrupting chemical also for mammals, including humans.
Additional studies supporting the endocrine disrupting properties of octylphenol in mammals can be found below in [1], [2], [3], [4].	See response to COM 24 in General comments
and response to COM 17 in this section	
			See also:
Comment 14 attachment (Annex A_references_Octylphenol.doc).doc		
13	2011/10/13	National NGO
Swedish Society for Nature Conservation (SSNC)
Sweden	[Page 155] Final conclusion stating that Octylphenol is not a "substance with ED properties of strong potency for the mammalian system". 

It is acknowledged in the dossier that there are several studies that indicate that octylphenol is an endocrine disruptor in mammals with "inherent potential for being toxic for reproduction" [page 153]. Taking into consideration the evidence pointing at octylphenol as an endocrine disruptor, we therefore believe it should be clearly stated that Octylphenol with the available evidence is to be considered an endocrine disrupting chemical also for mammals, including humans.

Additional studies supporting the endocrine disrupting properties of octylphenol in mammals can be found below in [1], [2], [3], [4]

Annex A - Studies not included in the German Annex XV dossier:

1.  Bonefeld-Jorgensen et al 2007; Endocrine-Disrupting Potential of Bisphenol A, Bisphenol A Dimethacrylate, 4-n-Nonylphenol, and 4-n-Octylphenol in Vitro: New Data and a Brief Review; 2007, Environ Health Perspect. 2007 December; 115(S-1): 69–76. 

2.  Christian & Gillies 1999; Developing hypothalamic dopaminergic neurones as potential targets for environmental estrogens; 1999, Journal of Endocrinology 160: R1-R6 

3.  Hernandez-Rodriguez et al 2007; Multigenerational study of the hepatic effects exerted by the consumption of nonylphenol- and 4-octylphenol- contaminated drinking water in Sprague–Dawley rats; 2007,  Environmental Toxicology and Pharmacology 23: 73-81 

4.  Sweeney et al, 2000; Maternal Exposure to Octylphenol Suppresses Ovine Fetal Follicle-Stimulating Hormone Secretion, Testis Size, and Sertoli CellNumber; 2000, Endocrinology 141: 2667-2673	See response to COM 24 in General comments
and response to COM 17 in this section	
12	2011/10/13	International NGO
ClientEarth
Belgium	•	[Page 155] Final conclusion stating that Octylphenol is not a "substance with ED properties of strong potency for the mammalian system". 
o	It is acknowledged in the dossier that there are several studies that indicate that octylphenol is an endocrine disruptor in mammals with "inherent potential for being toxic for reproduction" [page 153]. Taking into consideration the evidence pointing at octylphenol as an endocrine disruptor, we therefore believe it should be clearly stated that Octylphenol with the available evidence is to be considered an endocrine disrupting chemical also for mammals, including humans.
o	Additional studies supporting the endocrine disrupting properties of octylphenol in mammals can be found below in:
1.  Bonefeld-Jorgensen et al 2007; Endocrine-Disrupting Potential of Bisphenol A, Bisphenol A Dimethacrylate, 4-n-Nonylphenol, and 4-n-Octylphenol in Vitro: New Data and a Brief Review; 2007, Environ Health Perspect. 2007 December; 115(S-1): 69–76. 
2.  Christian & Gillies 1999; Developing hypothalamic dopaminergic neurones as potential targets for environmental estrogens; 1999, Journal of Endocrinology 160: R1-R6 
3.  Hernandez-Rodriguez et al 2007; Multigenerational study of the hepatic effects exerted by the consumption of nonylphenol- and 4-octylphenol- contaminated drinking water in Sprague–Dawley rats; 2007,  Environmental Toxicology and Pharmacology 23: 73-81 
4.  Sweeney et al, 2000; Maternal Exposure to Octylphenol Suppresses Ovine Fetal Follicle-Stimulating Hormone Secretion, Testis Size, and Sertoli CellNumber; 2000, Endocrinology 141: 2667-2673
5.  Ademollo et al 2008; Ademollo, N.; Ferrara, F.; Delise, M.; Fabietti, F.; and Funari, E. 2008. Nonylphenol and octylphenol in human breast milk. Environment International. 34: 984-987	See response to COM 24 in General comments
and response to COM 17 in this section	
11	2011/10/13	National NGO
Czech Republic	Page 155 - Final conclusion stating that Octylphenol is not a "substance with ED properties of strong potency for the mammalian system".

It is acknowledged in the dossier that there are several studies that indicate that octylphenol is an endocrine disruptor in mammals with "inherent  potential for being toxic for reproduction" [page 153]. Taking into consideration the evidence pointing at octylphenol as an endocrine disruptor, we therefore believe it should be clearly stated that Octylphenol with the available evidence is to be considered an endocrine disrupting chemical also for mammals, including humans.
Additional studies supporting the endocrine disrupting properties of octylphenol in mammals:
Bonefeld-Jorgensen et al 2007; Endocrine-Disrupting Potential of Bisphenol A, Bisphenol A Dimethacrylate, 4-n-Nonylphenol, and 4-n-Octylphenol in Vitro: New Data and a Brief Review; 2007, Environ Health Perspect. 2007 December; 115(S-1): 69­76.
Christian & Gillies 1999; Developing hypothalamic dopaminergic neurones as potential targets for environmental estrogens; 1999, Journal of Endocrinology 160: R1-R6
Hernandez-Rodriguez et al 2007; Multigenerational study of the hepatic effects exerted by the consumption of nonylphenol- and 4-octylphenol-contaminated drinking water in Sprague­Dawley rats; 2007, Environmental
Toxicology and Pharmacology 23: 73-81
Sweeney et al, 2000; Maternal Exposure to Octylphenol Suppresses Ovine Fetal Follicle-Stimulating Hormone Secretion, Testis Size, and Sertoli CellNumber; 2000, Endocrinology 141: 2667-2673	See response to COM 24 in General comments
and response to COM 17 in this section	
9	2011/10/12	Member State
Belgium	p.98 More recent and relevant publications describing different types of stable and transient reporter gene assays using fish cells (e.g. RTG-2, PLHC-1) with different type of constructs are not included in table 26 of the report. Several papers report both EC50 values for estrogen receptor binding (range 133 – 1938 nM) as well as potency data for OP through comparison with E2 (range 0.0013 – 0.006). It is recommended to include also in table 26 recent work on fish reporter assays (Ackermann et al., 2002; Rutishauser et al., 2004; Cosnefroy et al., 2009). 

Ackermann G.E., Brombacher E. & Fent K. (2002). Development of a fish reporter gene system for the assessment of estrogenic compounds and sewage treatment plant effluents. Environ. Toxicol. Chem. 21 (9): 1864–1875.

Rutishauser B.V., Pesonen M., Escher B.I., Ackermann G.E., Aerni H.-R., Suter M.J.-F. & Eggen R.I.L. (2004). Comparative analysis of estrogenic activity in sewage treatment plants effluents involving three in vitro assays and chemical analysis of steroids. Environ. Toxicol. Chem. 23 (4): 857–864, 2004

Cosnefroy A., Brion F., Guillet B., Laville N., Porcher J.M., Balaguer P. & Aït-Aïssa S. (2009). A stable fish reporter cell line to study estrogen receptor transactivation by environmental (xeno)estrogens. Toxicology in vitro 23: 1450-1454.

Editorial comments: 
Heading in table 26 on vitellogenin should be corrected. It is rather measurement of the VTG protein, than the VTG gene expression which is measured in presented assays. 
	Thank you for the new studies. They were carefully analysed. Potency described in these tests was in the same range as described in table 26.  As the dossier did not aim at providing an exhaustive analysis of in vitro data and new results did not change the overall conclusion, they were not included in the dossier. 	
7	2011/10/12	Member State
Norway	Cf. attached document	Thank you for your comments which were used to revise the dossier accordingly. With respect to the Klimisch code scoring, the study by Karels et al remained score 1 as the relevant part of the study was performed with a sufficient number of replicates (10 replicates with one male and female). The study by Anderson et al was changed to score 2 due to the missing information on test concentrations. The phrase “starting at” was removed from the dossier. Please note that – with the exception of P.reticulata - all overall conclusions were based on statistically significant effects and thus the overall conclusion does not change. Effects for which no statistics was provided remained in the dossier as they provided additional information on the mode of action in a weight of evidence approach.	
			See also:
Comment 7 attachment (Norwegian comments on OP SVHC dossier.doc).doc		
5	2011/10/12	National NGO
Bund für Umwelt und Naturschutz Deutschland (BUND) e.V. - Friends of the Earth Germany
Germany	[Page 155] Final conclusion stating that Octylphenol is not a "substance with ED properties of strong potency for the mammalian system". 

It is acknowledged in the dossier that there are several studies that indicate that octylphenol is an endocrine disruptor in mammals with "inherent potential for being toxic for reproduction" [page 153]. Taking into consideration the evidence pointing at octylphenol as an endocrine disruptor, we therefore believe it should be clearly stated that Octylphenol with the available evidence is to be considered an endocrine disrupting chemical also for mammals, including humans.

Additional studies supporting the endocrine disrupting properties of octylphenol in mammals can be found below in [1], [2], [3], [4]	See response to COM 24 in General comments
and response to COM 17 in this section	
			See also:
Comment 5 attachment (Annex to BUND comments on octylphenole dossier.doc).doc		
4	2011/10/12	International NGO
Center for International Environmental Law (CIEL)
United States	[Page 155] Final conclusion stating that Octylphenol is not a "substance
with ED properties of strong potency for the mammalian system".
> * It is acknowledged in the dossier that there are several studies that
> indicate that octylphenol is an endocrine disruptor in mammals with
"inherent
> potential for being toxic for reproduction" [page 153]. Taking into
> consideration the evidence pointing at octylphenol as an endocrine
disruptor,
> we therefore believe it should be clearly stated that Octylphenol with the
> available evidence is to be considered an endocrine disrupting chemical
also
> for mammals, including humans.
> * Additional studies supporting the endocrine disrupting properties of
> octylphenol in mammals can be found below in [1], [2], [3], [4], [5], [6]

Annex A - Studies not included in the German Annex XV dossier:
1.  Bonefeld-Jorgensen et al 2007; Endocrine-Disrupting Potential of
Bisphenol A, Bisphenol A Dimethacrylate, 4-n-Nonylphenol, and
4-n-Octylphenol in Vitro: New Data and a Brief Review; 2007, Environ Health
Perspect. 2007 December; 115(S-1): 69­76.
2.  Christian & Gillies 1999; Developing hypothalamic dopaminergic neurones
as potential targets for environmental estrogens; 1999, Journal of
Endocrinology 160: R1-R6
3.  Croteau, 2008; Chronic Exposure of Rana pipiens Tadpoles to UVB
Radiation and the Estrogenic Chemical 4-tert-octylphenol; 2008, Journal of
Toxicology and Environmental Health, Part A, 71: 134-144
4.  Hernandez-Rodriguez et al 2007; Multigenerational study of the hepatic
effects exerted by the consumption of nonylphenol- and 4-octylphenol-
contaminated drinking water in Sprague­Dawley rats; 2007,  Environmental
Toxicology and Pharmacology 23: 73-81
5.  Martin-Skilton et al 2006; Evidence of endocrine alteration in the red
mullet, Mullus barbatus from the NW Mediterranean; Environmental Pollution
Volume 141, Issue 1, May 2006, Pages 60-68
6.  Sweeney et al, 2000; Maternal Exposure to Octylphenol Suppresses Ovine
Fetal Follicle-Stimulating Hormone Secretion, Testis Size, and Sertoli
CellNumber; 2000, Endocrinology 141: 2667-2673	See response to COM 24 in General comments
and response to COM 17 in this section	
3	2011/10/12	International NGO
WECF - Women in Europe for a Common Future
Germany	[Page 155] Final conclusion stating that Octylphenol is not a "substance with ED properties of strong potency for the mammalian system". It is acknowledged in the dossier that there are several studies that indicate that octylphenol is an endocrine disruptor in mammals with "inherent potential for being toxic for reproduction" [page 153]. Taking into consideration the evidence pointing at octylphenol as an endocrine disruptor, we therefore believe it should be clearly stated that Octylphenol with the available evidence is to be considered an endocrine disrupting chemical also for mammals, including humans.
Additional studies supporting the endocrine disrupting properties of octylphenol in mammals can be found below in [1], [2], [3], [4]	See response to COM 24 in General comments
and response to COM 17 in this section	
1	2011/10/10	Trade union
European Trade Union Confederation
Belgium	4-(1,1,3,3-tetramethylbutyl)phenol, (4-tert-Octylphenol)is included in the Trade Union priority List for REACH authorisation.
see http://www.etuc.org/a/6023
	Thank you for this information	

PART II: Comments and responses to comments on uses, exposures, alternatives and risks

Specific comments on use, exposure, alternatives and risks
Number	Date	Submitted by (name, Organisation/MSCA)	Comment	Response	
20	2011/10/13	Industry or trade association
European Council for Alkylphenols and Derivatives (CEPAD)
Belgium	9.0 	As stated under Article 57 scientific evidence of probable serious effects to human health or the environment is necessary to classify a compound as a SVHC; the very low estrogenic potency and  low environmental occurrence and concentrations do not support a conclusion of OP of probable and serious environmental effects from this compound.
Relates to Part II, Section 7 of the Annex XV Report 
Categorization as SVHC should be reserved for substances for which the weight-of-evidence supports a finding of probable and serious effects.  As discussed previously, OP, at most, has weakly estrogenic activity and the available data do not support a conclusion that this compound is of equivalent concern to a CMR Cat. 1A or 1B, PBT or vPvB compound.  As discussed below, concentrations of OP in European waters are significantly below NOEC and LOEC values for adverse apical effects found in studies on OP in aquatic species.  In addition, studies that examine estrogenically active substances in wastewater treatment plant effluent found that if detected, OP and alkylphenols generally, contributed only minimally to the aggregate estrogenicity. 

9.1    Concentrations of OP in European surface waters do not support concern for
          probable and serious effects in aquatic species

As part of the Water Framework Directive, surface water concentrations of OP and numerous compounds have been measured in various European waterways (DG ENV, 2009). Concentrations of OP ranging up to 25,000 g/L were reported. These higher values are impossible as they exceed the aqueous solubility of OP of 7000 μg/L (IUCLID, 2010). There likely were errors made in the handling of the samples, the transcription of units, or analytical interferences may have been present. Rather than use the erroneous water concentrations, the median and upper 90th percentile concentrations better represent the waterways of Europe. The median and upper 90th percentile concentrations are 0.05 and 0.25 μg/L, respectively. These concentrations are well below all NOEC and LOEC from the screening, short term reproductive, and life cycle studies with OP. These data provide further “scientific evidence” that OP does not cause “probable serious effects” to the environment that are equivalent to CMR Cat. 1A and 1B, PBT, or vPvB substances.

9.2 	Contribution of OP to Measured Estrogenicity of Effluents or Surface Waters 
Numerous studies have examined the extent to which measurable estrogenic activity in wastewater treatment plant (WWTP) effluents is attributable to natural and synthetic compounds (e.g., E2 and EE2) or to other constituents (e.g., OP or other alkylphenols). 
Desbrow et al. (1998) used a sample fractionation system to isolate fractions of WWTP effluent samples that were tested for estrogenicity using a YES assay. Estrogenic fractions were found to contain natural and synthetic hormones including E2, estrone, and EE2. OP and other alkylphenolic compounds were not detectable in the estrogenic fractions. 
Most studies employed in vitro screening assays such as YES assays and concentration measurements from whole effluents to assess relative contributions to estrogenicity. Korner et al. (1999) and Aerni et al. (2004) reported that the contribution of NP in German WWTP effluents was <3% and 2%, respectively, of the total estrogenicity. In WWTP effluents in the US, Drewes et al. (2005) reported that nearly all of the estrogenic activity was attributable to natural and synthetic hormones. Similar findings have been reported for WWTP effluents in Australia, New Zealand, and China, with estrogenic contributions from OP of 0.04 to 3.9% (Leusch et al., 2006; Jin et al., 2007; Lu et al., 2011). 
	
9.3	OP is a chemical intermediate and its uses, emission and exposure profile do not support concern for environmental or human exposures of concern. 

In discussing the use and exposures of OP, the Annex XV Report notes that OP is a component in coatings, printing inks and some sorts of paints and as a tackifier in the production of rubber products.  It should be noted that OP is not used directly in these uses. Rather it is reacted further (i.e. as an ethoxylate or resin) before being used in these applications. 

In Table 4 (p. 159) of the Annex XV Report, a comparison of maximum, mean and median concentration shows single outliers indicative of a dispersive misuse of OP that is not representative for the general environment in Europe. The Report expresses concern that wide dispersive uses resulting in emission to wastewater might be important for OP. However, it should be noted that the alkylphenols industry does not support the use of OP or its derivatives in dispersive uses. 

	Please note that the SVHC identification is based on inherent properties and the possible effects a substance may have due to these properties. Thus a PEC/PNEC calculation is considered not appropriate

For (9.1) 
The concentration of 25000 µg/l refers to the concentration in water as whole with determination of concentration on each separate phase. The value represents the highest concentration found in a single phase. Having in mind the log Kow of 4-tert-octylphenole the substance will primarily adsorb on the suspended organic particles in the water body as stated in section 3.2.1. The concentration of 4-tert-octylphenole in the whole water body without separation of the single phases is indeed lower (1.08 µg/l). Median values proposed in the comments are also reported in the dossier..

For (9.2):
The comment is acknowledged but as stated above, exposure estimates are not considered in the dossier as they are not part of the SVHC identification

For(9.3):
We cannot fully share your opinion that there is no concern for humans and the environment. Your remark that OP as such is not an “active” component in products like paints and inks was confirmed to some extend after further review of the Exposure Scenarios submitted with the Chemical Safety Reports. Nevertheless free 4-tert-octylphenol is still present as impurity in the resins (up to 5%) and ethoxylates (up to 1%, depending on the grade of ethoxylation) and ethoxylates are known to degrade to OP in the environment.
We can not share your opinion that dispersive uses of octylphenole-ethoxylates are not supported by the alkylphenols industry, because the registration dossiers for 4-tert-octylphenole mention the use of octylphenole-ethoxylates for the production of paints, coatings and adhesives which will become available for wide dispersive uses by professional endusers and consumers (please see public section “identified uses” in ECHA Chem database for registered substances).	
			See also:
Comment 20 attachment (CEPAD-APERC Comments on OP SVHC (Oct. 13, 2011).zip).zip
		
19	2011/10/13	Industry or trade association
Alkylphenols & Ethoxylates Research Council
United States	9.0 	As stated under Article 57 scientific evidence of probable serious effects to human health or the environment is necessary to classify a compound as a SVHC; the very low estrogenic potency and  low environmental occurrence and concentrations do not support a conclusion of OP of probable and serious environmental effects from this compound.
Relates to Part II, Section 7 of the Annex XV Report 
Categorization as SVHC should be reserved for substances for which the weight-of-evidence supports a finding of probable and serious effects.  As discussed previously, OP, at most, has weakly estrogenic activity and the available data do not support a conclusion that this compound is of equivalent concern to a CMR Cat. 1A or 1B, PBT or vPvB compound.  As discussed below, concentrations of OP in European waters are significantly below NOEC and LOEC values for adverse apical effects found in studies on OP in aquatic species.  In addition, studies that examine estrogenically active substances in wastewater treatment plant effluent found that if detected, OP and alkylphenols generally, contributed only minimally to the aggregate estrogenicity. 

9.1    Concentrations of OP in European surface waters do not support concern for
          probable and serious effects in aquatic species

As part of the Water Framework Directive, surface water concentrations of OP and numerous compounds have been measured in various European waterways (DG ENV, 2009). Concentrations of OP ranging up to 25,000 μg/L were reported. These higher values are impossible as they exceed the aqueous solubility of OP of 7000 g/L (IUCLID, 2010). There likely were errors made in the handling of the samples, the transcription of units, or analytical interferences may have been present. Rather than use the erroneous water concentrations, the median and upper 90th percentile concentrations better represent the waterways of Europe. The median and upper 90th percentile concentrations are 0.05 and 0.25 μg/L, respectively. These concentrations are well below all NOEC and LOEC from the screening, short term reproductive, and life cycle studies with OP. These data provide further “scientific evidence” that OP does not cause “probable serious effects” to the environment that are equivalent to CMR Cat. 1A and 1B, PBT, or vPvB substances.
9.2 	Contribution of OP to Measured Estrogenicity of Effluents or Surface Waters 
Numerous studies have examined the extent to which measurable estrogenic activity in wastewater treatment plant (WWTP) effluents is attributable to natural and synthetic compounds (e.g., E2 and EE2) or to other constituents (e.g., OP or other alkylphenols). 
Desbrow et al. (1998) used a sample fractionation system to isolate fractions of WWTP effluent samples that were tested for estrogenicity using a YES assay. Estrogenic fractions were found to contain natural and synthetic hormones including E2, estrone, and EE2. OP and other alkylphenolic compounds were not detectable in the estrogenic fractions. 
Most studies employed in vitro screening assays such as YES assays and concentration measurements from whole effluents to assess relative contributions to estrogenicity. Korner et al. (1999) and Aerni et al. (2004) reported that the contribution of NP in German WWTP effluents was <3% and 2%, respectively, of the total estrogenicity. In WWTP effluents in the US, Drewes et al. (2005) reported that nearly all of the estrogenic activity was attributable to natural and synthetic hormones. Similar findings have been reported for WWTP effluents in Australia, New Zealand, and China, with estrogenic contributions from OP of 0.04 to 3.9% (Leusch et al., 2006; Jin et al., 2007; Lu et al., 2011). 
	9.3	OP is a chemical intermediate and its uses, emission and exposure profile do not support concern for environmental or human exposures of concern. 

In discussing the use and exposures of OP, the Annex XV Report notes that OP is a component in coatings, printing inks and some sorts of paints and as a tackifier in the production of rubber products.  It should be noted that OP is not used directly in these uses. Rather it is reacted further (i.e. as an ethoxylate or resin) before being used in these applications. 

In Table 4 (p. 159) of the Annex XV Report, a comparison of maximum, mean and median concentration shows single outliers indicative of a dispersive misuse of OP that is not representative for the general environment in Europe. The Report expresses concern that wide dispersive uses resulting in emission to wastewater might be important for OP. However, it should be noted that the alkylphenols industry does not support the use of OP or its derivatives in dispersive uses. 	See response to comment 20	
			See also:
Comment 19 attachment (CEPAD-APERC Comments on OP SVHC (Oct. 13, 2011).zip).zip		
18	2011/10/13	National NGO
Danish Ecological Council
Denmark	In the dossier, important data are excluded as applicable, such as effects induced through subcutaneous or interperitoneal routes of exposure (page 154). These data should instead be included in the overall weight of evidence for effect on human health, considering that these routes of exposure could indicate effects induced through dermal exposure or in some cases inhalation.
Lately (Feb. 2011), a paper shows that spermatogenesis is blocked at the round spermatid stage in high-dose exposed mice, subcutaneously injected with 4-tert-octylphenol. This further indicates, that data from subcutaneous exposed animals are important to include.
Since 4-tert-octylphenol is found in a wide variety of consumer articles such as clothing, we argue that both dermal and inhalation exposure routes should be taken into consideration. It says in the dossier “results summarized in (Environment Agency UK, 2008) showing that octylphenols were found in nearly all analyzed imported colored textile articles”.	Thanks for the comments. They are appreciated and will provide valuable input in further discussions.	
17	2011/10/13	National NGO
Réseau Environnement Santé
France	Page [156] Exposure:

Despite a reference in the dossier to 
Santillo D.; Labunska I.; Davidson H.; Johnston P.; Strutt M.; Knowles O.: Consuming
Chemicals. Hazardous Chemicals in house dust as an indicator of chemical exposure in the home.
Greenpeace Research Laboratories, Technical Note 2003, 01/2003.
showing the widespread presence of alkylphenols including OP in housedusts, inhalation as a probable route of exposure has not been taken into account in the dossier and no studies have been presented with this route of administration. 

Dermal exposure is not considered in the dossier with a reference to an evaluation made in 2001, Community Strategy for Endocrine Disrupters [COM(2001)262] even if octylphenol is assumed to have a high bioavailability through dermal exposure route. That the dermal route is neglected is rather striking in the light of the UK Environment Agency test of octylphenol in textile 2008 [page157] "Showing that octylphenols were found in nearly all analyzed imported colored textile articles" 

Both dermal and inhalation exposure routes should be taken into consideration due to the fact that octylphenol is found in a wide variety of consumer articles such as clothing.

[Page 163] Human biomonitoring: 
The dossier refers to several human monitoring studies showing that octylphenol can be found in blood, urine and breast milk. 2 additional studies (7 and 8) in our uploaded annex), not included in the Annex XV dossier, show similar result of octylphenol found in human body while study 9 adresses the daily intake of German babies.	Thank you for the information. The studies should be considered in further discussions on human exposure to 4-tert-Octylphenole. For the study by Ademollo et al. (reference 7) it should be clarified, whether the determined substance is 4-tert-Octylphenol, CAS-No. 140-66-9 or 4-Octylphenol, CAS-No. 1806-26-4.  	
			See also:
Comment 17 attachment (Annex RES octylphenol.pdf).pdf		
16	2011/10/13	Member State
United Kingdom	The German c.a. has quoted from three reports produced by the UK under its Co-ordinated Chemicals Risk Management Programme (a risk evaluation report, targeted assessment for textile washing, and risk reduction strategy). We do not have any further data to offer. However, we would like to offer a word of warning about Pollution Inventory data (as cited from the European Environment Agency). From experience, we know that some site operators make simplistic estimates of releases (sometimes based on analytical detection limits), so the veracity of the data would need to be considered.

It is worth noting that the exposure scenarios in our assessments will pre-date those in the REACH registrants’ Chemical Safety Assessments. There seems to be no review of the latter, which is a gap.
	Thank you for your comment.

We are aware that for many substances information from the PRTR is only the result of a rough estimation. However, the number of sites not only in UK but also across Europe indicates that emissions from 4-tert-OP seem to be – together with other uses - the result of a wide dispersive use of products containing ethoxylates made from 4-tert-OP.

The exposure scenarios of the registrants were checked in order to obtain some information on tonnages for the single registered uses and where briefly discussed in the confidential part of the SVHC dossier. A detailed examination on emissions to the environment was not conducted due to the timeframe and because this is not inside the scope of identification process for a SVHC.

Information from the registrants´Chemical Safety Assessments including exposure scenarios is briefly discussed in the confidential part of the dossier. 
	
15	2011/10/13	Other contributor
CALEB MANAGEMENTS SERVICES LIMITED
United Kingdom	The comments below are submitted by CALEB on behalf of the 
European Phenolic Resins Association

7.1.2 Use

Page 156 “as a component in phenolic resins”

For phenolic resins, 4-tOP is not a component but a monomer that is chemically reacted with formaldehyde and possibly other reactants to form 4-tOP based phenolic resins. 4-tOP is only contained at residual concentration in the resins and can not re-form upon degradation of the resins. The content of residual 4-tOP ranges from 0.1-5% depending on the resin grade.

Page 156 “component in coatings, printing inks…..” 

4-tOP is not used in its neat form but in the reacted form of phenolic resins. 
Depending on the grade and amount of phenolic resin used, the content of free 4-tOP in those applications is very low.

Page 156  “as tackifier”

4-tOP is not used in its neat form for the production of rubber products but in the reacted form of phenolic resins. Residual 4-tOP contents in the resin grades used are typically from 1-5%. 

7.1.3.1 Environment

Page 158 ”Measured releases “ 

Monitoring has been conducted in 2006 at two relevant resin manufacturing sites and the PECs (Clocal) based on monitoring results were below the PNEC of 0.632 µg/L.
A further decrease can be predicted for the following years.

Page 159 “Together with the emissions from disposal of non-hazardous waste and incineration of non-hazardous waste in total 2039 kg of 4-tert-octylphenol and his ethoxylates were emitted in the UK area in 2008.“

It is unclear, how 4-tOP emissions could result from incineration of non-hazardous waste since 4-tOP is combustible under the conditions of controlled incineration.

	Thank you for your comments.

The information on the chemical reaction for producing of 4-tert-OP based phenolic resins was acknowledged and will be respected in the further work.

7.1.3.1:
p. 158/159:
Thank you for this additional information. However, this does not change the overall conclusion that emission of 4-tert-octylphenol from resin manufacturing sites as well as other uses might be a relevant source of OP for the environment. Further information would be needed to assess the assignability of these data to the overall situation in Europe. 
The PRTR-register refers to two facilities for incineration or disposal of waste emitting 53.3 kg 4-tert-octylphenol or its ethoxylates in 2008. Incineration of non-hazardous waste is the main activity of these two sites which also serve as urban waste-water treatment plants. The available data does not allow a differentiation if the emissions result from the incineration activities or the treatment of urban waste-water.	
14	2011/10/13	International NGO
ChemSec
Sweden	Page [156] Exposure:
Inhalation as a probable route of exposure has not been taken into account in the dossier and no studies have been presented with this route of administration. 
Dermal exposure is not considered in the dossier with a reference to an evaluation made in 2001, Community Strategy for Endocrine Disrupters [COM(2001)262] even if octylphenol is assumed to have a high bioavailability through dermal exposure route. That the dermal route is neglected is rather striking in the light of the UK Environment Agency test of octylphenol in textile 2008 [page157] "Showing that octylphenols were found in nearly all analyzed imported colored textile articles" 
Both dermal and inhalation exposure routes should be taken into consideration due to the fact that octylphenol is found in a wide variety of consumer articles such as clothing.
[Page 163] Human biomonitoring: The dossier refer to several human monitoring studies showing that octylphenol can be found in blood, urine and breast milk. An additional study [5], not included in the Annex XV dossier, show similar result of octylphenol found in breast milk. 	See response to National NGO
Réseau Environnement Santé
France	
			See also:
Comment 14 attachment (Annex A_references_Octylphenol.doc).doc		
13	2011/10/13	National NGO
Swedish Society for Nature Conservation (SSNC)
Sweden	Page [156] Exposure:

Inhalation as a probable route of exposure has not been taken into account in the dossier and no studies have been presented with this route of administration. 

Dermal exposure is not considered in the dossier with a reference to an evaluation made in 2001, Community Strategy for Endocrine Disrupters [COM(2001)262] even if octylphenol is assumed to have a high bioavailability through dermal exposure route. That the dermal route is neglected is rather striking in the light of the UK Environment Agency test of octylphenol in textile 2008 [page157] "Showing that octylphenols were found in nearly all analyzed imported colored textile articles" 

Both dermal and inhalation exposure routes should be taken into consideration due to the fact that octylphenol is found in a wide variety of consumer articles such as clothing.

[Page 163] Human biomonitoring: The dossier refer to several human monitoring studies showing that octylphenol can be found in blood, urine and breast milk. An additional study [5], not included in the Annex XV dossier, show similar result of octylphenol found in breast milk.

Annex A - Studies not included in the German Annex XV dossier:

5.  Ademollo et al 2008; Ademollo, N.; Ferrara, F.; Delise, M.; Fabietti, F.; and Funari, E. 2008. Nonylphenol and octylphenol in human breast milk. Environment International. 34: 984-987	See response to National NGO
Réseau Environnement Santé
France	
12	2011/10/13	International NGO
ClientEarth
Belgium	•	Page [156] Exposure:
o	Inhalation as a probable route of exposure has not been taken into account in the dossier and no studies have been presented with this route of administration. 
o	Dermal exposure is not considered in the dossier with a reference to an evaluation made in 2001, Community Strategy for Endocrine Disrupters [COM(2001)262] even if octylphenol is assumed to have a high bioavailability through dermal exposure route. That the dermal route is neglected is rather striking in the light of the UK Environment Agency test of octylphenol in textile 2008 [page157] "Showing that octylphenols were found in nearly all analyzed imported colored textile articles" 
o	Both dermal and inhalation exposure routes should be taken into consideration due to the fact that octylphenol is found in a wide variety of consumer articles such as clothing.
•	[Page 163] Human biomonitoring: The dossier refer to several human monitoring studies showing that octylphenol can be found in blood, urine and breast milk. An additional study [5], not included in the Annex XV dossier, show similar result of octylphenol found in breast milk. 
	See response to National NGO
Réseau Environnement Santé
France	
11	2011/10/13	National NGO
Czech Republic	Page 156 - Exposure:
Inhalation as a probable route of exposure has not been taken into account in the dossier and no studies have been presented with this route of administration.

Dermal exposure is not considered in the dossier with a reference to an evaluation made in 2001, Community Strategy for Endocrine Disrupters [COM(2001)262] even if octylphenol is assumed to have a high bioavailability through dermal exposure route. That the dermal route is neglected is rather striking in the light of the UK Environment Agency test of octylphenol in textile 2008 

Page 157 -  "Showing that octylphenols were found in nearly all analyzed imported colored textile articles"

Both dermal and inhalation exposure routes should be taken into consideration due to the fact that octylphenol is found in a wide variety of consumer articles such as clothing.

Page 163 - Human biomonitoring: The dossier refer to several human monitoring studies showing that octylphenol can be found in blood, urine and breast milk. An additional study [Ademollo et al 2008; Ademollo, N.; Ferrara, F.; Delise, M.; Fabietti, F.; and Funari, E. 2008. Nonylphenol and octylphenol in human breast milk. Environment International. 34: 984-987], not included in the Annex XV dossier, show similar result of octylphenol found in breast milk.	See response to National NGO
Réseau Environnement Santé
France	
8	2011/10/12	Industry or trade association
ETRMA - European Tyre & Rubber Manufacturers' Association
Belgium	One of the main uses of octylphenol is in the production of para-tert-Octylphenol(PTOP) based resins, which are used as tackifiers and for metal to rubber bonding applications both in tyres and technical general rubber goods. These PTOP based resins are produced by reaction of PTOP with other substances, for example formaldehyde, and are polymeric in nature. Thus the large majority of the PTOP used in these applications is in a polymerized form and no longer exists as PTOP. These resins do however typically contain free PTOP monomer impurity of between 1-5%.
Specifically, PTOP based resins are sometimes used in tyre compounds, typically in the carcass plies or steel belts, in order to ensure adequate adhesion, and the production of
a high quality product. By promoting the adhesion between tyre components, PTOP based resins are essential to the product safety and performance.
Tyres are high tech products and are extensively tested to make sure that they fulfill the required safety standards. Accordingly, any changes to tyre formulations are treated
very cautiously and require extensive research and development work to assure products’ safety performance.
According to company data on annual use of PTOP based resins, and tyre production,and assuming an average content of 3% free impurity PTOP in the resins, it has been calculated that an average EU tyre contains between 0,007 % and 0,012 % of PTOP.
This takes a highly conservative assumption that all impurity PTOP would remain even after the extensive processing and high temperatures of the manufacturing processes.
The above calculation demonstrates the incorrect assumption in the UK Environment Agency 2005 report, mentioned in the German Annex XV dossier, that tyres contain 0.3% PTOP.
In addition to the fact that PTOP is only present as an impurity, reduced exposure to the ingredients of rubber compounds occurs immediately after mixing with rubber
polymers. The bioavailability of the ingredients from this stage on is strongly reduced as a result of:
- high capacity of the polymeric matrix to entangle substances, due to the physicochemical interactions established between the polymeric network and the individual substances;
- compact characteristics of the compound, which lead to the incorporation of the majority of the ingredients and make them unavailable to the environment and humans.

End of Life tyre applications could provide a potential local exposure point for PTOP,
however the aforementioned aspects of the very low concentration and lack of
bioavailability make it extremely unlikely that this would be a significant source of PTOP.
For instance, as reported in the ADEME study*, from an ecotoxicological viewpoint, the
results obtained show that the nature of the percolates likely to infiltrate into the
ground underlying the artificial turf sports surface proves to be without impact on the
aquatic environment in the short and medium term (standardized tests carried out on
the first percolates and repeated several times during the year).

* (ADEME / ALIAPUR / FIELDTURF TARKETT (2007) Environmental and health evaluation
of the use of elastomer granulates (virgin and from used tyres) as filling in thirdgeneration
artificial turf
http://www.aliapur.fr/media/files/RetD_new/Synthetic_turf_-_Environmental_Study_Report.pdf	Thank you for your support. Any further information to composition of tyres like your one is welcome.

As stated in the dossier resins used as tackifiers in the production of tyres typically contain residues of free 4-tert-octylphenole in concentrations up to 4%. The value maximum of 0.3% 4-tert-OP in tyres is cited from the UK Risk Evaluation Report (2005) is a result of personal communication between the responsible authority and the European Council for Alkylphenols and Derivates (CEPAD). CEPAD is cited in the UK report “...that the maximum figure for the percentage of resin in rubber used for tyres is 10%. This results in a maximum concentration of free 4-tert-octylphenole in tyres of 0.3%.” Because there is less public information on
substances and amounts used for tyre production this value was chosen for the dossier because to represent a worst case. Your information will be respected during RMO development and any further information from your side is welcome. As stated in your comment the concentration of 0.012% refers to an average EU tyre – nevertheless there might be imported tyres from outside the EU which may contain free 4-tert-OP in higher concentrations.

The conclusion on reduced bioavailability due to inclusion in the polymeric matrix cannot be shared, because there are several examples of substances with similar or more complex chemical structure used in the production of rubber goods that are unhinged from these kind of products.

Thank you for sharing information on emissions from artificial turf sports surfaces. Nevertheless there also several studies mentioned in the Annex XV-Dossier indicating that leaching of residual free 4-tert-OP from rubber crumb will occur.	
			See also:
Comment 8 attachment (20111003 Octhylphenol - ETRMA response to consultation - final version.pdf).pdf		
6	2011/10/12	Individual
Belgium	Re: Sec. 7.1

Its Annex XV candidate dossier states that 4-tert-octylphenol (OP) is expected to degrade when released to water post-wastewater treatment.  The following abstract indicates OP may be more refractory than that in aquatic environments, making all the way into drinking water at concentrations your dossier says other studies find OP in drinking water at.  Unfortunately the abstract does not give what concentrations of OP were found downstream of WWTP release, but perhaps the full paper does.  So I ask that you check if it reports any detection of OP in freshwaters, and see if they exceed the freshwater PECs the dossier gives.

I strongly encourage you to encourage & require more listing of chemicals of "equivalent concern" for consideration as SVHC!
--
Environ Sci Technol. 2001 Aug 1;35(15):3201-6.
Determination of endocrine-disrupting phenolic compounds and estrogens in surface and drinking water by HRGC-(NCI)-MS in the picogram per liter range.
Kuch HM, Ballschmiter K.
Source
Department of Analytical and Environmental Chemistry, University of Ulm, Germany.
Abstract
A method for the analysis of phenolic estrogenic active compounds in surface and drinking water in the picogram per liter range is described. Besides the widely used monomer bisphenol A, 4-tert-octylphenol [4-(1,1,3,3-tetramethylbutyl)phenol] and the technical isomer mixture of 4-nonylphenol; phenolic steroid hormones such as the endogenous estrogens estrone, 17alpha-estradiol, and 17beta-estradiol; and the exogenous estrogen 17alpha-ethinylestradiol were determined in water at the 20-200 pg/L level. Water samples from 1 to 5 L were extracted by solid-phase extraction (SPE) on a cartridge system containing LiChrolut EN as sorbent. The phenols and steroids were converted into their pentafluorobenzoylate esters in an extractive derivatization reaction. The derivatives were then determined by high-resolution gas chromatography with negative chemical ionization mass spectrometric detection (HRGC-(NCI)-MS) in the selected ion mode (SIM). All results were also confirmed by HRGC with electron capture detection (ECD). This highly sensitive and specific method gives a limit of detection (LOD) of 20 pg/L for bisphenol A and 4-tert-octylphenol in drinking water samples and 50 pg/L in STW effluent, respectively. The LODs for technical 4-nonylphenol, 17alpha-ethinylestradiol, and other estrogens are in the range of 50 pg/L in drinking water to 200 pg/L in STW effluent, respectively. In all river water samples in southern Germany, bisphenol A was found in concentrations ranging from 500 pg/L up to 16 ng/L, 4-nonylphenol was from 6 up to 135 ng/L, and the steroids were from 200 pg/L up to 5 ng/L. In drinking water, bisphenol A was found in concentrations ranging from 300 pg/L to 2 ng/L, 4-nonylphenol was from 2 to 15 ng/L, 4-tert-octylphenol was from 150 pg/L to 5 ng/L, and the steroids were from 100 pg/L to 2 ng/L. Mean recoveries over the whole analytical protocol, measured in bidistilled water, generally exceeded 70%. These results indicate that environmental endocrine-disrupting estrogens are not completely removed in the process of sewage treatment but are carried over into the general aquatic environment. After ground passage, they can eventually be found in drinking water.
==		
5	2011/10/12	National NGO
Bund für Umwelt und Naturschutz Deutschland (BUND) e.V. - Friends of the Earth Germany
Germany	Page [156] Exposure:

Inhalation as a probable route of exposure has not been taken into account in the dossier and no studies have been presented with this route of administration. 

Dermal exposure is not considered in the dossier with a reference to an evaluation made in 2001, Community Strategy for Endocrine Disrupters [COM(2001)262] even if octylphenol is assumed to have a high bioavailability through dermal exposure route. That the dermal route is neglected is rather striking in the light of the UK Environment Agency test of octylphenol in textile 2008 [page157] "Showing that octylphenols were found in nearly all analyzed imported colored textile articles" 

Both dermal and inhalation exposure routes should be taken into consideration due to the fact that octylphenol is found in a wide variety of consumer articles such as clothing.

[Page 163] Human biomonitoring: The dossier refer to several human monitoring studies showing that octylphenol can be found in blood, urine and breast milk. An additional study [5], not included in the Annex XV dossier, show similar result of octylphenol found in breast milk. 	See response to National NGO
Réseau Environnement Santé
France	
			See also:
Comment 5 attachment (Annex to BUND comments on octylphenole dossier.doc).doc		
4	2011/10/12	International NGO
Center for International Environmental Law (CIEL)
United States	* Page [156] Exposure:
> * Inhalation as a probable route of exposure has not been taken into
account
> in the dossier and no studies have been presented with this route of
> administration. 
> * Dermal exposure is not considered in the dossier with a reference to an
> evaluation made in 2001, Community Strategy for Endocrine Disrupters
> [COM(2001)262] even if octylphenol is assumed to have a high
bioavailability
> through dermal exposure route. That the dermal route is neglected is
rather
> striking in the light of the UK Environment Agency test of octylphenol in
> textile 2008 [page157] "Showing that octylphenols were found in nearly all
> analyzed imported colored textile articles"
> * Both dermal and inhalation exposure routes should be taken into
> consideration due to the fact that octylphenol is found in a wide variety
of
> consumer articles such as clothing.
* [Page 163] Human biomonitoring: The dossier refer to several human
monitoring studies showing that octylphenol can be found in blood, urine and
breast milk. An additional study [7], not included in the Annex XV dossier,
show similar result of octylphenol found in breast milk.	See response to National NGO
Réseau Environnement Santé
France	
3	2011/10/12	International NGO
WECF - Women in Europe for a Common Future
Germany	Page [156] Exposure:
Inhalation as a probable route of exposure has not been taken into account in the dossier and no studies have been presented with this route of administration. 
Dermal exposure is not considered in the dossier with a reference to an evaluation made in 2001, Community Strategy for Endocrine Disrupters [COM(2001)262] even if octylphenol is assumed to have a high bioavailability through dermal exposure route. That the dermal route is neglected is rather striking in the light of the UK Environment Agency test of octylphenol in textile 2008 [page157] "Showing that octylphenols were found in nearly all analyzed imported colored textile articles" 
Both dermal and inhalation exposure routes should be taken into consideration due to the fact that octylphenol is found in a wide variety of consumer articles such as clothing.
[Page 163] Human biomonitoring: The dossier refer to several human monitoring studies showing that octylphenol can be found in blood, urine and breast milk. An additional study [5], not included in the Annex XV dossier, show similar result of octylphenol found in breast milk. 	See response to National NGO
Réseau Environnement Santé
France	

Attachments

Comment number	Public attachment	
5	
Comment 5 attachment (Annex to BUND comments on octylphenole dossier.doc).doc	
7	
Comment 7 attachment (Norwegian comments on OP SVHC dossier.doc).doc	
8	
Comment 8 attachment (20111003 Octhylphenol - ETRMA response to consultation - final version.pdf).pdf	
14	
Comment 14 attachment (Annex A_references_Octylphenol.doc).doc	
17	
Comment 17 attachment (Annex RES octylphenol.pdf).pdf	
19	
Comment 19 attachment (CEPAD-APERC Comments on OP SVHC (Oct. 13, 2011).zip).zip	
20	
Comment 20 attachment (CEPAD-APERC Comments on OP SVHC (Oct. 13, 2011).zip).zip	
23	
Comment 23 attachment (CHEM Trust Position on EDC Criteria - Sept11.pdf).pdf	
24	
Comment 24 attachment (CHEM Trust Position on EDC Criteria - Sept11.pdf).pdf	
26	
Comment 26 attachment (OP_EDC_SVHC-tt.pdf).pdf