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

Substance name: Butyl 4-hydroxybenzoate
CAS number: 94-26-8
EC number: 202-318-7

The substance is proposed to be identified as meeting the following SVHC criteria set out in Article 57 of the REACH Regulation: Endocrine disrupting properties (Article 57(f) – human health)

Disclaimer: Comments provided during the consultation are made available as submitted by the commenting parties. It was in the commenting parties own responsibility to ensure that their comments do not contain confidential information. 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. 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, submitter type, country)	Comment	Responses	
5383
2020/04/16	Health and Environment Alliance (HEAL),
International NGO,
Belgium		Thank you for your support and the information provided in your detailed comments with several references proposed for consideration.  

The reference from WHO, (World Health Organisation, Human Biomonitoring: Facts and figures, 2015) discusses parabens in general and no data on butylparaben is provided. Therefore, this reference is not included in the revised document. 

The reference proposed from Hass et al (2012) is based on a Danish project analysed by national experts who raised the concern of butylparaben having endocrine disrupting properties. Conclusions are based on proposed Danish criteria for EDs (to date not internationally agreed) and is considered a “historical” background document and thus not included in the reference list. 

The reference from the European Commission (Call for data on potential EDs, May 2019) does not provide data that are directly relevant for the hazard identification of butylparaben as a SVHC and thus not included in the supporting document. 

The reference from La Merrill et al. (2020) is considered a conceptual paper on ED substances and does not discuss butylparaben. Therefore, this paper is not included in the reference list. 

The reference proposed from Iribarne-Durán et al. (2019) presents an overview of the presence of BPA and parabens, including butylparaben, in neonatal intensive care unit along with two in vitro assays (E-Screen and PALM). 
Data from these in vitro screening assays are pooled which makes it impossible to determine the magnitude of each substance related to the observed effects. Therefore, the direct link between butylparaben and the effects observed is not clear. 
The overview of the presence of each of the substances is in our view relevant for the exposure of butylparaben. However, exposure and uses of the substance is not relevant for the hazard identification of butylparaben as a SVHC. Therefore, this reference is not included in the supporting document. 

The reference from Hagai et al. (2017) is a general analysis of trends in sperm counts and does not discuss impacts directly related to butylparaben. Therefore, this reference is not included in the reference list. Additionally, the consequences of impaired fertility are addressed in section 6.1.2, 6.1.3 and Annex II in the supporting document.

References from Brunel University (2013), WHO Europe (2006), Health and Environment Alliance (2013), Trasande et al. (2016) and Fertility Europe (2017) does not discuss butylparaben. They are considered relevant for general ED and infertility discussions and thereby are not directly relevant for the identification of butylparaben as a SVHC. 
	
		Attachment: 5383_HEAL comments-Butylparaben.pdf

5405
2020/04/17	Health & Safety Executive,
National Authority,
United Kingdom	In vitro an in vivo mechanistic studies (4.5.1.2 – page 13/14)

Endocrine Activity In Vitro
The available in vitro information on the endocrine activity of butyl paraben comes from non-standard published studies.  There is limited information on design, conduct and reporting of these studies, such that it is difficult to conduct an independent evaluation.

In these studies, there is some in vitro evidence that butyl paraben can bind to and activate the oestrogen receptor.  There is limited evidence that butyl paraben may have some antiandrogenic and antiestrogenic capacity, and interfere with steroidogenesis.  There is no information on the relative potency in these assays of butyl paraben compared with reference compounds. Therefore, it is not possible to infer how potent butyl paraben may be.

We also note butyl paraben appears to be rapidly and extensively metabolised in vivo.  As none of these in vitro studies apparently included an exogenous metabolising system, the relevance of any conclusions for the in vivo situation is uncertain.

Endocrine Activity in vivo
There are no standard studies available to investigate the in vivo endocrine activity of butyl paraben.  The only available information comes from a number of non-standard uterotrophic assays & published studies, which have a number of limitations, in terms of design, conduct and reporting, when compared to modern OECD TGs.

There are 8 uterotrophic assays available, 7 of which employed the sub-cutaneous route of exposure, which is not relevant for industrial chemicals.  Butyl paraben was reported to have tested negative in the only oral uterotrophic assay, conducted in rats.  There is no further information available, such as positive control performance or doses employed.

There are no Hershberger assays available conducted with butyl paraben.  Evidence for antiandrogenic activity in vivo was provided by a single positive Hershberger assay, conducted with the analogue propyl paraben.

It is stated that AGD was decreased in two rat studies, in which dams were dosed orally with butyl paraben at doses of up to 1000 mg/kg/day (GD7-PND 21/22).  However, there is no information on the magnitude of the changes, nor on the presence or degree of general toxicity/changes in weight-gain of the offspring.  Without this information, it is not possible to judge the toxicological significance of these findings.
It is noted that some of these testicular effects occurred at doses which also caused a decrease in testosterone and an increase in oestrogen levels.  However, it is unclear whether the changes in hormone levels caused the reported testicular effects.

Changes in some hormone levels (decreased testosterone and increased oestrogen) were reported in intact male rats following pubertal or adult exposure.  These changes may indicate endocrine activity, in vivo.  However, there is no information on general toxicity to inform on the toxicological significance of this finding.

Overall, there is no clear evidence in vivo, in intact animals, employing a relevant route of exposure, which allows the conclusion that butyl paraben has in vivo endocrine activity.

Endocrine Activity in vivo and adversity (4.5.1.3 pages 14-16)
There are no standard studies available to provide information on the potential of butyl paraben to cause reproductive toxicity in experimental animals.

In some oral studies in rats (prenatal/perinatal dosing) it was reported that butyl paraben caused testicular toxicity (decreased sperm count/motility, and inconsistent changes in testicular weight) (1/3 oral dosing studies).  In adult rats, 2/3 studies reported a decrease in sperm numbers and 1/3 studies reported decreased sperm motility and abnormal testicular histopathology.  However, there is very limited information available for any of these studies, including no information on the degree/extent of any general toxicity or magnitude of the reported changes.  Without this information it is not possible to judge the toxicological significance of these findings.
It is noted that some of these testicular effects occurred at doses which also caused a decrease in testosterone and an increase in oestrogen levels.  However, it is unclear whether the changes in hormone levels caused the reported testicular effects.

We note there is limited in vitro information that butyl paraben has some endocrine activity from non-standard tests (oestrogenicity/antioestrogenicity, antiandrogenicity and interference with steroidogenesis).

However, there is no convincing corroborating evidence from a number of in vivo studies; for example butyl paraben tested negative in a non-standard uterotrophic assay.  Furthermore, there is no evidence from the pre-natal oral dosing studies for aberrant development of the male reproductive tract, which might be anticipated from the in vitro endocrine activity data.  Overall, there is no coherence between the in vitro studies and in vivo studies.
	Thank you for your comments. The conclusion that butylparaben exerts ED-related effects is considered robust and relies on a weight of evidence approach taking into account inter alia the Guidance for the identification of endocrine disruptors in the context of Regulations (EU) No 528/2012 and (EC) No 1107/2009 (ECHA/EFSA 2018). Non-guideline information was evaluated for quality, relevance and reliability. In general, the same principles for relevance and reliability apply as for literature data from standard studies as also prescribed by the ECHA Guidance on information requirements and chemical safety assessment, Chapter R4: evaluation of available information.

The internationally agreed WHO/IPCS definition of EDs does not include potency considerations or requirements. Furthermore, there are several indications that for EDs it is the time of exposure during pregnancy and/or early life-stages that matters (exposure during critical time windows) rather than the dose. In the light of this, potency considerations are not required for the identification of 57(f) for butylparaben.

It is clear from human studies that despite rapid metabolism, the parent substance is measureable in human serum samples, and therefore, the endocrine activities of the parent substance is relevant. 
In addition, as noted in the section ‘Lines of evidence’ (Annex I) some metabolites are also tested in vitro. As mentioned in the dossier, in vitro studies performed by Gonzales et al. (2018) and Watanabe et al. (2013) found that butylparaben and the butylparaben metabolite 3-hydroxy n-butyl 4-hydroxybenzoate showed similar estrogen receptor agonistic response patterns to estrogen. The hydroxylated isomers of butylparaben (n-butyl-3-hydroxybenzoate and n-butyl-2-hydroxybenzoate) also activated the estrogen receptor, however, to a lower degree than butylparaben. This information is considered sufficient evidence for estrogen receptor activation.

The conclusion that butylparaben exerts ED-related effects is considered robust following the same argumentation as stated above regarding non-standard studies.

The subcutaneous route has been used in a number of studies on butylparaben because the daily dose can be controlled with greater accuracy. 
Compared to the oral route, subcutaneous administration bypasses the digestive barrier and most importantly the enterohepatic circulation and first-pass metabolism. In addition, in rodents, and to a lesser extent in humans, the hepatic metabolism capacity in newborns is limited, resulting in a reduced hepatic first-pass effect. 
The subcutaneous route of exposure is therefore considered relevant for the purpose of the present analysis that is hazard identification and MoA considerations, in particular when perinatal exposure is considered.
Information from an additional oral study conducted in rats by Vo et al. (2010) is now included in the revised document. Although this study shows no effect on uterus weight, increased thickness of endometrium was reported in rats orally exposed to butylparaben. 
Information on uterine histology was not provided in the oral uterotrophic assay that tested negative with butylparaben. 
Changes of uterine histology indicate some activation of the estrogen pathway and the study by Vo et al. (2010) is thus included in the supporting evidence for estrogenic mode of action of butylparaben.  

The Hershberger assay conducted with propylparaben is reasonable to justify expected effects of butylparaben and additional references on the similarity of the substances are now provided in section 4.5.1.2. This gap-filling approach leads to reduced experimental testing, as there is no need to test every target substance. 
Additionally, reported anti-androgenic effects are considered supporting evidence, due to less consistent data. This points to the possibility of multiple modes of action.

Information on the magnitude of the changes and changes in weight-gain of offspring are now provided in the detailed study information in Annex III for the references Zang et al. (2014) and Boberg et al. (2016). 

Requirements of the WHO/IPCS definition and its agreed European interpretation (JRC, 2013) involve demonstration that the adverse effects are plausibly caused by an alteration of the endocrine function. There is therefore no requirement to understand the whole mode of action in question. The mode of action analysis is presented and discussed in section 4.5.2 and Annex II. 

It is unclear which studies are referenced in your comment. Section 4.5.1.2 does not present nor discuss studies that report changes in hormone levels (decreased testosterone and increased oestrogen) in intact male rats following pubertal/adult exposure. 

The scientific evidence of probable serious effects to human health is considered clear.

Same argumentation regarding non-guideline studies as stated above.

Available information regarding oral rat studies is now provided in the detailed study information (Annex III).

For each of the effects included in the prepared dossier, we consider that the available experimental data provides sufficient evidence to identify the adversity.
Based on weight of evidence assessment of strengths and limitations of all studies available, induction by butylparaben of the adverse effects described cannot be refuted. In absence of conclusive human studies but considering the relevance for humans of the effects observed on animals, these effects can be considered as “presumed” for humans.
	
5414
2020/04/17	Cosmetics Europe,
Industry or trade association,
Belgium	Attachment: 5414_ButylParaben_additional info.zip
	Answers are provided below. 

Specific comments on the justification
Number / Date	Submitted by (name, submitter type, country)	Comment	Responses	
5379
2020/04/16	Finland,
Member State	We thank Danish CA for this proposal. The FI CA acknowledges that SVHC identifications based on the equivalent level of concern (ELoC) in relation to the Art. 57 (f) are made on a case by case basis. The SVHC proposal is based on comprehensive and systematic assessment of the data presented in peer reviewed publications. 

The Finnish CA notes that a Risk Management Option Analysis (RMOA) conclusion document has not been published on the ECHA website for this substance. 

We further note that in contrast to previously identified SVHC´s based on ED properties for human health, the adverse effects presented in this Annex XV report (reduced sperm counts and quality) have not been acknowledged by harmonized classification.	Thank you for your comments.

We take note of this comment although it is not directly relevant for SVHC identification of butylparaben which is based on the intrinsic properties of the substance. Further risk management options will be addressed at the next steps if deemed necessary.
However, as mentioned in the prepared dossier, the ED properties of the substance was discussed by the Endocrine Disruptor Expert Group on 7 October 2019. There was broad agreement by the experts that the available information seems to be sufficient to identify the substance as an ED for human health.

Considerations on classification is noted. 
However, we agree that many substances identified as endocrine disruptors for human health are usually classified or notified as reproductive toxicants. Currently, this is not the case for butylparaben. The concern for endocrine activity of butylparaben was raised by national experts in a project evaluating substances with endocrine disrupting properties (Hass et al. 2012). The data presented in the prepared dossier is not analysed with the aim of classifying butylparaben.

Reference: Hass, U., Christiansen, S., Petersen, M. A., Boberg, J., Andersson, A-M., Skakkebæk, N. E. Bjerregaard, P. (2012). Evaluation of 22 SIN List 2.0 substances according to the Danish proposal on criteria for endocrine disrupters. DTU Food.	
5382
2020/04/16	Environment Agency Austria,
Other contributor,
Austria	The Environment Agency Austria supports the identification of butyl 4-hydroxybenzoate (commonly referred to as butylparaben) as a substance of very high concern in accordance with Article 57(f) of Regulation (EC) 1907/2006 (REACH). There is scientific evidence of probable serious effects to human health due to the substance’s endocrine disrupting properties, which give rise to an equivalent level of concern to those substances listed in points (a) to (e) of Article 57 REACH.
Butylparaben leads to reduced sperm count and quality (number of normal sperm) particularly at perinatal exposure. The evidence for endocrine activity is sufficient with evidence for estrogenic activity, altered steroidogenesis and androgen receptor antagonism. The link between the adverse effects and the endocrine mode of action is plausible.
If deemed adequate, the study of Leppert et al. (2020) could be discussed in the dossier, as for prenatal butylparaben exposure in humans a positive association is observed to overweight within the first eight years of life with a stronger trend in girls in this study - likely due to an epigenetic modification in the neuronal Pro-opiomelanocortin (POMC) enhancer 1 leading to a reduced hypothalamic POMC expression.
Reference: Leppert, B., Strunz, S., Seiwert, B. et al Maternal paraben exposure triggers childhood overweight development. Nat Commun 11, 561 (2020). https://doi.org/10.1038/s41467-019-14202-1 
	Thank you for support and information provided.

Reference from Leppert et al. (2020) is now added in the reference list and analysed in Annex V. It is considered a robust study as it includes mechanistic follow up studies in rats. The mechanism suggested is thereby plausible and seems relevant to the effects observed in humans. This indicate absorption and some activity of butylparaben during pregnancy. However, as the study presented no data relevant to evaluation of reproductive effects of butylparaben, it is not discussed further in the supporting document.	
5383
2020/04/16	Health and Environment Alliance (HEAL),
International NGO,
Belgium	The Health and Environment Alliance (HEAL) thanks Denmark for the proposal to identify Butyl 4-hydroxybenzoate (hereafter butylparaben) as a substance of very high concern (SVHC) under REACH article 57(f). We fully support this proposal, which is documented by a very strong and well-structured dossier.
Our detailed comments can be found in the attached document.	As mentioned in Table 1, we thank you for your support.	
		See the corresponding embedded attachment in table 1 of Part I: 5383_HEAL comments-Butylparaben.pdf		
5390
2020/04/17	Norway,
Member State	The Norwegian Environment Agency thanks the Danish Environmental Protection Agency (DK-EPA) for preparing the SVHC proposal for butylparaben due to the criteria set out in REACH Article 57 (f), based on endocrine disruptors for human health. The regulation of endocrine disruptors is of high priority in Norway both for human health and the environment.

We support the assessment regarding the intrinsic properties of butylparaben as well as the equivalent level of concern (ELOC) assessment performed by DK-EPA.
The adverse health effect reported following exposure to butylparaben was a reduction in sperm count and sperm quality (number of normal sperm). This was observed in experimental animal studies where rodents were exposed during the perinatal period, however uncertainties for the induction of these effects following exposure during the prepubertal/adult exposure periods were observed. These effects are considered severe since similar effects in humans could result in sub- and infertility. The effects occur later in life and a long time after the exposure during the perinatal period has occurred. Supporting evidence of adverse effects were related to an altered AGD (ano-genital distance), altered sperm motility and reports of histological changes in testis in some studies. Further, some studies also reported effects on the female reproductive system following perinatally exposure to butylparaben.

The adverse effects on sperm production is acknowledged. However, as the effects are related to reproductive toxicity it should be considered to submit a proposal for harmonised classification for effects on sexual function and fertility to ECHA for the assessment of the adversity of the effects on male reproduction.

We also consider that there is sufficient evidence to conclude that butylparaben acts via endocrine mode(s) of action. The evidence was shown to be strongest for estrogenic activity in in vitro and in vivo studies, however, some studies also showed altered steroidogenesis and androgen receptor (AR) antagonism.
As regards the equivalent level of concern it is supported that infertility can have an adverse effect on the propagation of the species, as well as having an impact on the quality of life. Further, no safe concentration level can be derived from the available experimental animal studies on the reproductive effects reported to be via an endocrine mode of action. It is therefore supported that exposure to butylparaben is considered to give rise to an equivalent level of concern to those of other substances listed in points (a) to (e) of Article 57 REACH.	Thank you for your comments and support.

Your considerations on a proposal for harmonised classification is noted. However, the data presented in the prepared dossier is not analysed with the aim of classifying butylparaben. The concern for endocrine activity was raised by national experts in a project evaluating substances with endocrine disrupting properties (Hass et al. 2012). The 57 (f) identification of butylparaben for human health was partly the outcome of this project.

Reference: Hass, U., Christiansen, S., Petersen, M. A., Boberg, J., Andersson, A-M., Skakkebæk, N. E. Bjerregaard, P. (2012). Evaluation of 22 SIN List 2.0 substances according to the Danish proposal on criteria for endocrinedisrupters. DTU Food.	
5397
2020/04/17	ANSES,
National Authority,
France	The identification of Butyl 4-hydroxybenzoate (EC 202-318-7) as an SVHC due to its endocrine properties relevant for human health as proposed by Danish authorities is supported.	Thank you for your support.	
5398
2020/04/17	ChemSec,
International NGO,
Sweden	ChemSec supports the identification of butylparaben as an endocrine disrupting chemical and SVHC. We added the substance to the SIN List in 2011 and since then there is even more evidence of its negative impacts on human health. The dossier presents well especially the evidence of butylparaben decreasing sperm quality and sperm number. 

A few more studies are even available since the dossier was published, eg.
https://doi.org/10.1016/j.envpol.2019.112957 
It especially concerning that this chemical is so commonly used that it is very often found in human urine, blood and breast milk. Apart from affecting sperm quality it has also other impacts, ie it has been associated with overweight in offspring. In our opinion also Propylparaben should be identified as an EDC along with butylparaben as they are very similar in both structure, use and negative impacts on human health.	Thank you for support and information provided.

We have noted that butylparaben is on the SIN List. However, reference to this list is not included in the supporting document, as it does not provide additional information on the endocrine disrupting properties of butylparaben related to human health. 

Reference from Maske et al. (2020) is now added in the reference list and quoted in section 4.5.1.3 and Annex I, III and IV. In addition, we added studies by van Meeuwen et al., 2008, and Vo et al., 2010.

We agree that propylparaben is structurally related to butylparaben and the intrinsic properties of both substances seem to be similar. However, identification of butylparaben as an ED is in our view a first step for developing future dossiers for additional parabens.
	
5405
2020/04/17	Health & Safety Executive,
National Authority,
United Kingdom	Before concluding on ELoC and whether Candidate Listing is justified, it should be considered whether the WHO ED (for human health) criteria have been met:

“An endocrine disruptor is an exogenous substance or mixture that alters functions(s) of the endocrine system and consequently causes adverse health effects in an intact organism, or its progeny, or (sub)populations”

From the available information there is no clear, convincing evidence that butyl paraben has any in vivo endocrine activity when administered to intact experimental animals by a relevant route of exposure (oral, dermal or inhalation routes only).

As such, we consider that the information provided in the SVHC report is insufficient for us to conclude that butyl paraben meets the WHO criteria of an ED for human health and therefore its identification as an SVHC is currently not supported.

Substance evaluation would allow for a tiered testing approach following the OECD conceptual framework to allow clarification of any ED properties and we consider this could be a first step.
	Thank you for your comment. However, we do consider the information provided in the prepared dossier as sufficient to conclude that butylparaben meets the WHO criteria of an ED for human health, including convincing evidence of endocrine activity in vivo. 
In the updated dossier, information is added from one additional in vivo study showing effects on sperm parameters in rats (Maske et al., 2020).

We agree with the statement that substance evaluation would allow for additional testing of the substance. However, we consider the available evidence to be sufficient for the identification of butyl paraben as a SVHC based on REACH article 57(f). 
Therefore, we consider initiation of a substance evaluation needless. 	
5407
2020/04/17	CHEM Trust Europe,
International NGO,
Germany	CHEM Trust supports the identification of butyl 4-hydroxybenzoate (butylparaben) as SVHC based on the endocrine disrupting properties and in accordance with REACH Art. 57(f).

The Annex XV dossier convincingly demonstrates that butyl paraben has endocrine disrupting properties, warranting the identification as endocrine disruptor in accordance with the ECHA/EFSA guidance for the identification of endocrine disrupting biocides and pesticides and the WHO/IPCS definition of an endocrine disruptor as interpreted by the JRC Endocrine Disruptors Expert Assessment Group.

The presented analysis of the in vitro and in vivo mechanistic studies revealed that there is sufficient evidence for estrogenicity of butylparaben. Moreover, supporting evidence for altered steroidogenesis and anti-androgenicity is included. It is shown that the mode of action of butylparaben is based on “EATS-mediated adversity”; likely acting via multiple modes of action.

Likewise, the analysis revealed that there is sufficient evidence of adverse effects in vivo (decreased sperm count and decreased number of normal sperm) and this is supported by evidence of endocrine activity in vivo (altered hormone levels) and adverse effects (altered anogenital distance, decreased sperm motility, altered testicular histopathology).

Furthermore, no safe concentration/level can be derived from the available data on adverse reproductive effects via endocrine modes of action.

It should be noted that given the EATS-mediated adversity and in particular the impacts on fertility/reproduction it is highly likely that butylparaben is also an ED for the environment and should be treated as such. Studies have shown that parabens may have estrogenic effects in fish (Pedersen KL, Pedersen SN, Christiansen LB, Korsgaard B, Bjerregaard P. The preservatives ethyl-, propyl- and butylparaben are oestrogenic in an in vivo fish assay. Pharmacol Toxicol 2000;86:110–3).

Butylparaben has been included in ChemSec`s SIN list since 2011 as an endocrine disruptor with estrogenic and antiandrogen activity, affecting sperm function and reproductive organs among others which had been detected in various (bio)monitoring studies.
	Thank you for your support.

We acknowledge that the ED related effects of butylparaben may be wider than those described in the prepared dossier. However, environmental data on butylparaben was not reviewed and thus not included in the document. The identification of butylparaben as an ED for human health is in our view a first step for developing future dossiers for additional concerns.	
5408
2020/04/17	Sweden,
Member State	We thank Denmark for this proposal.
We agree, based on the provided data in the dossier, that this substance has endocrine disrupting properties and that there is scientific evidence of probable serious effects to human health which give rise to an equivalent level of concern, according to the criteria of article 57(f) in REACH, to those of other substances listed in points (a) to (e).

We would like to propose to consider the newly published data from NTP (NIEHS, USA) on reproductive and developmental effects (Link to NTP: https://manticore.niehs.nih.gov/cebssearch/paper/14506. Only raw data is currently provided.).	Thank you for your support and information provided. 

We are aware of the newly published data from NTP. However, we consider that it would be too premature to include it, as only raw data is available. The data from NTP will be relevant for evaluation when a full study report is available. 
Please find attached a note by the Research Group for Molecular and Reproductive Toxicology, Technical University of Denmark (DTU) regarding new NTP studies on butylparaben:   

5414
2020/04/17	Cosmetics Europe,
Industry or trade association,
Belgium	Butyl Paraben Annex XV Dossier (DK, Feb 2020)
COMMENTS BASED ON THREE KEY AREAS OF CONCERN (A to C)
Submitted with regard to:

Substance name: Butyl 4-hydroxybenzoate (Butylparaben)
EC Number: 202-318-7
CAS number: 94-26-8

The comments provided below are solely referring to n-butylparaben and arise from assessments independent from those of other substances, such as isobutylparaben

A. Evidence and its significance
Page 15 of Butyl Paraben Annex XV Dossier (DK, Feb 2020):
The epidemiological studies referred to (Aker et al, 2016, 2019a; Fernandez et al, 2016) do not support the claim in the Danish Annex XV dossier that “effects on human fertility and development have been described in studies that examined exposures in adulthood and during pregnancy and childhood in relation to health effects early or late in life”. The results described borderline trends in lower maternal testosterone levels associated with higher maternal urinary butyl paraben levels and a significant negative association between maternal urinary butyl paraben and sex hormone binding globulin, as well as a negative association between maternal urinary butyl paraben levels and maternal serum levels of oestradiol and the oestradiol/progesterone ratio. It is well accepted that a single urinary measurement at only a single time-point does not demonstrate an increased exposure to parabens. No information is provided about whether the urine samples were taken from a 24-hour urine collection or whether they were adjusted for urinary output (i.e., normalized to creatinine). In absence of this correction the concentration data are meaningless. In other words: the lower the volume of excreted urine, the higher would be the measured butyl paraben concentration.

In addition, association does not mean causal link. Also, there is a lack of biological plausibility between the alleged association between urinary butyl paraben and levels of different sex hormones which have a great variability and fluctuation.

Another study showed no association between placental butylparaben levels and congenital malformations of male genitalia (cryptorchidism and hypospadias). The fact that urinary residues of butylparaben were more frequently detected in cases versus controls does not provide supportive evidence for the hypothesis that butyl paraben may induce any endocrine-related adverse effect during pregnancy. Again, no information was provided about whether butyl paraben data were from a 24-hour urine collection or whether they were adjusted for urinary output (i.e., normalized to creatinine). Without this correction, the concentration data are meaningless. The lower the volume of excreted urine, the higher would be the measured butyl paraben concentration.
Overall, these studies provide no evidence of endocrine-related adverse effects from butyl paraben exposure in humans.

Page 12, 14, 16 of Butyl Paraben Annex XV Dossier (DK, Feb 2020):
The included rodent data do not demonstrate that butyl paraben induced effects on fertility related parameters (sperm count, sperm quality, testicular function) and development (birth parameters, growth, adverse effects on early reproductive development or male reproductive function). The studies outlined in the dossier are largely inconsistent in their results and do not show any pertinent adverse effect caused by butyl paraben. The inconsistencies between studies are explained to be probably due to different exposure periods, dose levels and measuring sensitivity. However, the inconsistent results rather confirm that butyl paraben with its very weak  endocrine activity, i.e. an estrogenic activity which is several magnitudes below 17-β-estradiol, is not able to induce relevant adverse endocrine-related effects in vivo.

There is no evidence of relevant adverse effects on reproductive and developmental parameters in rodents from state-of-the-art GLP studies such as Hoberman et al (2008), whereas academia non-GLP studies (Boberg et al, 2016; Kang et al, 2002; Zhang et al, 2014) claim to demonstrate weak effects which inconsistent with the negligible in vitro endocrine activity of butyl paraben. However, a detailed scientific review of the rodent studies by Boberg et al, 2016; Kang et al, 2002; Zhang et al, 2014 did not confirm the adverse effects claimed by the authors. Furthermore, several inconsistencies between the study results indicate the absence of a relevant in vivo effect by butyl paraben. For example, the Zhang et al (2014) and Boberg et al (2016) studies report a reduction of anogenital distance (AGD) in the offspring of pregnant rats given oral doses (gavage) of butyl paraben during GD7-PND21/22. This effect is not considered to be meaningful since this parameter is not well defined in humans or non-human primates compared to rodents and moreover, was not observed in various other studies even after subcutaneous injection (Kang et al, 2002; Taxvig et al, 2008; Hoberman et al, 2008). On the other hand, Kang et al (2002), Zhang et al (2014) and Boberg et al (2016) report a reduced epididymal sperm count in rats (at doses which very likely induced general toxicity after SC injection in DMSO, Kang et al 2002), also at doses where other studies (Taxvig et al, 2008; Hoberman et al, 2008) do not find this effect. Therefore, on the basis of all these remarkable inconsistencies the available data do not allow to conclude that there is sufficient evidence of adverse effect in vivo (decreased sperm count and decreased number of normal sperm), supported by evidence of endocrine activity in vivo (altered hormone levels) and adverse effects (lower mean anogenital distance, decreased sperm motility, altered testicular histopathology). If the reduction in sperm count in male offspring observed in a few studies is compared to historical control data, the conclusion would be “no relevant effect”. Likewise, with respect to the presented mechanism of action (MoA), such as a reduced testicular CYP19α1 (aromatase) expression in pre-pubertal (not in adult male rats!) leading to disrupted spermatogenesis (Boberg et al, 2016), whereas in Zhang et al (2014), the hypothesis was suggested that an overexpression of aromatase may lead to dysmorphic seminiferous tubules and disrupted spermatogenesis. However, no relevant data supporting this hypothesis were presented in any study to prove such a MoA of butyl paraben.

In conclusion, weighing the evidence is an inherent part of a scientific assessment. Therefore, on the basis of reliability, relevance and consistency, the available in vivo data provide no evidence of endocrine-related adverse effects from butyl paraben exposure in animals or humans.

B. Level of Exposure
Page 11 of Butyl Paraben Annex XV Dossier (DK, Feb 2020):
Metabolism experiments (Hoberman et al, 2008) indicate that butyl paraben is rapidly metabolized by non-specific esterases to p-hydroxybenzoic acid and butanol, neither of which is estrogenic. The rapid metabolism of parabens by esterases is responsible of the very low amount of systemically bioavailable unmetabolised paraben. This probably explains why these weakly estrogenic substances elicit no in vivo effects when administered by relevant exposure routes (i.e., topical and oral). Interestingly, rat studies after subcutaneous (SC) injection of high doses (Taxvig et al, 2008) did not show any adverse butyl paraben-related effects, although the SC injection bypasses the skin barrier and prevents the skin metabolism of butyl paraben. Therefore, it may be expected that subcutaneous administration may induce higher butyl paraben levels than those after oral exposure which would result in quick metabolisation by the liver.

C. ELoC and Safe Concentration
Page 20-22 of Butyl Paraben Annex XV Dossier (DK, Feb 2020):
Article 57(f) of REACH specifically requires scientific evidence of ‘probable serious effects’ to demonstrate an Equivalent Level of Concern (ELoC), but the claim by DK-EPA that butyl paraben meets this criterion is unfounded.

There is no evidence from in vivo data and only insufficient evidence from in vitro data that butyl paraben may exert adverse effects based on an endocrine-related mode of action, i.e. “EATS-mediated adversity”. The DK-EPA itself admits that there is insufficient data on adverse effects to conclude on the endocrine disruption (p. 21).

However, DK-EPA implies that no safe concentration/level can be derived from the available data on adverse reproductive effects via endocrine modes of action. In their opinion, two of the available studies show reduced sperm count or quality in perinatally exposed rats at the lowest tested dose, and no NOAEL can be determined for this endpoint. However, as outlined in section A., these studies do not support the view of DK-EPA. Large state-of-the-art GLP studies (Hoberman et al, 2008) demonstrate that the effects published from academia non-GLP studies are not relevant and that the NOAEL after 8-week oral dietary exposure of male rats to butyl paraben can be established at the top dose of 10,000ppm (>1000 mg/kg bw/day). At this dose no adverse effect occurred on organ weights, histopathology of reproductive tissues, sperm production, motility, morphology and reproductive hormone levels.

On the basis of the absence of toxicity at this high dose, a safe concentration/level can be derived for the relevant routes of potential human exposure (oral and dermal). This is supported by the fact that butyl paraben is a very efficient preservative with a long history of safe use in cosmetic products.	Thank you for your comments. 
Responses are provided below:

The prepared dossier aims to summarise and transparently discuss the overall evidence that butylparaben has ED-mediated effects as well as its uncertainties as pointed out in your comment. We consider it important to base a SVHC identification of butylparaben on a critical scientific assessment. This effort for a balanced analysis should not be seen as a weakness of the dossier but definitely as its strength.
As specified in Annex I presenting the lines of evidence analysis, the epidemiological data are included as supporting evidence. As is often the case for epidemiological studies, the ability to identify cause and effect relationships is limited, particularly when fetal exposure is proposed to have led to adverse health outcomes in adulthood, i.e. decades later.

For details on urine samples, we refer to the data presented in Annex V. Urine samples were collected as ‘random spot’ or ‘timed spot’.  Specific gravity or creatinine were used as adjustment of urinary dilution and in a single study, osmolality was used.

The details on urine samples requested are available in Annex V.

As mentioned above, the epidemiological data are included as supporting evidence which is in line with the ECHA/EFSA ED guidance, page 21:  
“When assembling and assessing the line of evidence, any available epidemiological studies should be considered as supportive evidence for the evaluation of whether an ED is likely to have adverse effects for humans. However, they cannot be used to override or dismiss evidence of adversity found in laboratory studies, nor can they replace laboratory studies.”

It is acknowledged in the prepared dossier that divergent results are reported across studies for these endpoints and these discrepancies were discussed. Most of the effects seem to be sensitive to many factors across studies, including the level of doses, period of exposure, species and strain differences. Due to variations in the combination of these factors in the different experimental studies available, as well as to variations in the endpoints investigated and sometimes to experimental limitations, it is very difficult to capture the specific influence of each parameter on the effect. However, considering the respective limitations and strengths of the different studies available, the endocrine disrupting properties of butylparaben have been sufficiently established using a weight of evidence approach

With respect to GLP versus non-GLP studies, it is particularly important to note that no GLP studies were available examining adverse effects of perinatal exposure to butylparaben. The study by Hoberman et al. (2008) is not relevant in this respect.

With respect to adverse effect on AGD, the limitations are discussed in the dossier, and this is considered supporting evidence.

Findings of reduced epididymal sperm count was considered sufficient evidence of adverse effect of perinatal exposure. In the updated support document, information is added from one additional in vivo study showing effects on sperm parameters in rats (Maske et al., 2020).
The studies by Taxvig et al. (2008) and Hoberman et al. (2008) did not examine epididymal sperm count after perinatal exposure, and it is puzzling why these studies are referred in this comment as studies that “do not find this effect”. 

Comparison with in-study data is considered relevant, whereas historical control data are not available and no conclusion can be drawn in this regard.

One of the presented modes of action is “Altered steroidogenesis/increased estradiol level” (see Annex II). As summarised, some studies show an upregulated aromatase activity, whereas other studies show no increase in estradiol production, and therefore the evidence for this molecular initiating event is considered “moderate”. In contrast, the evidence for the molecular initiating event “Activation of estrogen receptor” is considered “high” (Annex II).   

Comments regarding use, exposure, alternatives, and risks, if relevant, may be considered at a later stages of the risk management process but is not relevant for the identification of the substance as a SVHC which is based on the hazard properties of the substance.

Responses regarding metabolism of butylparaben and subcutaneous administration, are provided in this consultation document (no. 5405). 

We agree that the substance has to show an Equivalent Level of Concern (ELoC) to fulfil the SVHC criteria according to Art 57(f) and the prepared dossier discusses in detail how butylparaben give rise to an equivalent level of concern to those of other substances listed in Art 57 points (a) to (e).  
In addition, as noted in the section 6.3 ‘Equivalent level of concern assessment’, endocrine disrupting (ED) properties of butylparaben relevant for human health include reduced sperm count and quality (number of normal sperm) as observed in rodent studies using perinatal exposure. Effects are irreversible and are shown to occur later in life after exposure in the perinatal period only. No safe concentration/level can be derived from the available data on adverse reproductive effects via endocrine modes of action. There is sufficient evidence to conclude that butylparaben acts via endocrine mode(s) of action according to a mode of action analysis including an evaluation of biological plausibility. These effects are considered severe as similar effects in humans could cause sub- and infertility. Based on the above, there is clear evidence that the substance is of an ELoC.

This statement regarding insufficient data on adverse effects only applies to pubertal/adult exposure. 
For perinatal exposure, the evidence on adverse effects is considered sufficient, as there is sufficient dose and temporal concordance between key events, and effects that are assumed relevant for humans.

In general, there are several indications that for substances with endocrine disrupting properties the time of exposure during pregnancy and/or early life-stages is crucial (exposure during critical time windows) rather than the dose.

Uses of butylparaben are not relevant to the hazard identification and MoA of the substance.

Responses are provided in the table in part II.	
		See the corresponding embedded attachment in table 1 of Part I: 5414_ButylParaben_additional info.zip		
5416
2020/04/17	European Environmental Bureau (EEB),
International NGO,
Belgium	The EEB supports the proposal by Denmark to identify butyl 4-hydroxybenzoate (butylparaben) as SVHC based on its endocrine effects for human health (REACH Article 57(f)). Prenatal exposure to butylparaben causes reduced quantity and quality of sperm later in life. The rapporteur demonstrated convincingly the plausibility of a causal link between the adverse effects and endocrine modes of action of butylparaben via estrogenic activity, androgenic antagonism and possibly altered steroidogenesis. The reduced quantity and quality of sperm after prenatal exposure leads to serious and irreversible adverse effects later in life.
In conclusion, the EEB agrees to the identification of butylparaben as substance of very high concern according to article 57(f) of REACH.	Thank you for your support.	
5417
2020/04/17	Netherlands,
Member State	The reason why the Annex XV dossier was submitted (PBT/vPvB, CMR, equivalent concern):
It is proposed to identify the substance as a substance of equivalent level of concern to those of other substances listed in points (a) to (e) of Article 57 of Regulation (EC) No 1907/2006 (REACH) according to Article 57(f) of REACH Regulation.

1. General Comments
NL agrees that the available information is sufficient to identify the substance as an endocrine disruptor for human health via EAS modalities. For the sake of completeness it could be considered to also include the test results of Toxcast in which the substance has been active in 111 out of 738 assays.
In the dossier it is mentioned that the ECHA/EFSA guidance is followed which means that also an environmental assessment should be included. We therefore do not agree with the statement that an environmental hazard assessment is not relevant for the identification of the substance as SVHC in accordance with Article 57 point (f) REACH due to its endocrine disrupting properties for human health. It could well be that other risk management measures are needed when the substance is identified as an endocrine disruptor for human health. We therefore would like to propose to rephrase this statement, as to say that the available and current information is not sufficient to draw a conclusion that the substance meets the criteria for an endocrine disruptor relevant for the environment.
	Thank you for your support and information provided.

Noted. We have now included reference to Toxcast data in the dossier.

In the dossier, it is mentioned that the ECHA/EFSA guidance is inter alia taken into account and not strictly followed as it primarily applies obligations under the Biocidal Product Regulation (BPR) and the Plant Protection Products Regulation (PPPR).

We acknowledge your considerations on substances with endocrine disrupting properties for human health as being relevant to the environment as well. However, environmental data for butylparaben was not reviewed and thus not included in the document. 
Experts in a national project evaluating substances with endocrine disrupting properties raised the concern for butylparaben exerting endocrine activity for human health (Hass et al. 2012). The 57 (f) identification of butylparaben for human health was partly the outcome of this project. The identification of butylparaben as an ED for human health is in our view a first step for developing future dossiers for additional concerns.

The statement mentioned is rephrased in the updated supporting document.

Reference: Hass, U., Christiansen, S., Petersen, M. A., Boberg, J., Andersson, A-M., Skakkebæk, N. E. Bjerregaard, P. (2012). Evaluation of 22 SIN List 2.0 substances according to the Danish proposal on criteria for endocrine disrupters. DTU Food.	

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, submitter type, country)	Comment	Responses	
5383
2020/04/16	Health and Environment Alliance (HEAL),
International NGO,
Belgium	See the corresponding embedded attachment in table 1 of Part I: 5383_HEAL comments-Butylparaben.pdf
	Responses are provided in table 1 of part 1.	
5414
2020/04/17	Cosmetics Europe,
Industry or trade association,
Belgium	See the corresponding embedded attachment in table 1 of Part I:5414_ButylParaben_additional info.zip	Thank you for your comment. However, your considerations regarding the importance of keeping a wide palette of preservatives are not directly relevant for the 57(f) identification of butylparaben. Your reference to the Cosmetic Product regulation 1223/2009/EC (the positive list concept) is related to uses and exposure of substances and thereby relevant for risk management. Risk management is not a part of the hazard identification of butylparaben as a SVHC. 

Additionally, it is noted that the attached safety report published by the CIR Panel (2019) seems to evaluate the risks of butylparaben along with other parabens used in cosmetics. Risk assessments are not considered directly relevant for the hazard identification of the substance. 

Comments regarding use, exposure, alternatives, and risks, if relevant, may be considered at later stages of the risk management process but is not relevant for the identification of the substance as a SVHC which is based on the hazard properties of the substance.