<b>Bijsluiter</b>. De hyperlink naar het originele document werkt niet meer. Daarom laat Woogle de tekst zien die in dat document stond. Deze tekst kan vreemde foutieve woorden of zinnen bevatten en de opmaak kan verdwenen of veranderd zijn. Dit komt door het zwartlakken van vertrouwelijke informatie of doordat de tekst niet digitaal beschikbaar was en dus ingescand en vervolgens via OCR weer ingelezen is. Voor het originele document, neem contact op met de Woo-contactpersoon van het bestuursorgaan.<br><br>====================================================================== Pagina 1 ======================================================================

<pre>Bisphenol A
Health-based recommendation on occupational exposure limits
To: the State Secretary of Social Affairs en Employment
No. 2019/04, The Hague, March 26, 2019
                                                            2 2
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<pre> Contents                                                                                                            Bisphenol A | page 2 of 51
 contents
      Samenvatting3                                                  3.3  The European Food Safety Authority (EFSA) (2015)                 15
                                                                      3.4  The Risk Assessment Committee (RAC) (2015)                       16
      Executive summary                                         6    3.5  The National Institute for Public Health and
                                                                           the Environment (RIVM) (2014, 2016)                              17
 01 Scope8                                                           3.6  Recent literature                                                17
      1.1   Background and objective                            9
      1.2   Committee and procedure                             9 04 Hazard assessment                                                    20
      1.3   Data                                                9    4.1  Evaluation of (non-)monotonic dose response relationships        21
                                                                      4.2  Hazard identification                                            22
 02 Uses, existing guidelines and standards                    11    4.3  Quantitative hazard assessment                                   24
      2.1   Uses                                               12    4.4  Skin notation                                                    25
      2.2   Current exposure limits for the working population 12    4.5  Groups with increased risk                                       26
      2.3   Classification                                     12    4.6  Conclusions and recommendation                                   26
 03 Selection of previous evaluations and recent                      Literature                                                           27
      literature13
      3.1   The Netherlands Health Council (1996)              14    Annex                                                                35
      3.2   The Scientific Committee on Occupational                  A    Recent literature                                                36
            Exposure Limits (SCOEL) (2013)                     14
1       Health Council of the Netherlands | No. 2019/04                                                    2                                  3
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<pre> Samenvatting                                                                                                                   Bisphenol A | page 3 of 51
 samenvatting                                                                                           een grenswaarde voor beroepsmatige
                                                                                                        blootstelling kan vaststellen.
 Op verzoek van de minister van Sociale Zaken         Daarnaast kan blootstelling aan bisfenol A leiden In 1996 heeft de Gezondheidsraad voor
 en Werkgelegenheid (SZW) heeft de                    tot schade aan de ogen, allergische huidreacties  beroepsmatige blootstelling een advieswaarde
 Gezondheidsraad de gezondheidskundige                en irritatie van de luchtwegen. Er bestaan        afgeleid van 10 milligram (mg) inhaleerbaar
 advieswaarde voor beroepsmatige blootstelling        binnen de EU beperkingen voor het gebruik van     bisfenol A per kubieke meter (m3) lucht, en een
 aan bisfenol A geactualiseerd.                       bisfenol A. Zo is het sinds 2011 verboden in      advieswaarde van 5 mg per m3 voor respirabel
                                                      flesjes voor babyvoeding en het gebruik in        bisfenol A. Momenteel geldt een grenswaarde
 Dit advies is tot stand gekomen in de Commissie      thermisch papier (voor kassabonnen) is sinds      van 2 mg inhaleerbaar bisfenol A per m3 lucht.
 Gezondheid en beroepsmatige blootstelling aan        2016 aan banden gelegd. Voor het gebruik in       Die grenswaarde is gebaseerd op een advies
 stoffen (GBBS). Op www.gezondheidsraad.nl            materialen die in contact komen met voedsel       van de Europese Scientific Committee on
 staat informatie over de taken van deze vaste        gelden maxima voor de hoeveelheid bisfenol A      Occupational Exposure Limits (SCOEL) uit
 commissie van de Gezondheidsraad. De                 die eruit vrij mag komen.                         2013.
 samenstelling van de commissie is te vinden
 achterin dit advies.                                 Gezondheidskundige advieswaarde                   Sinds 2013 zijn er tal van wetenschappelijke
                                                      Voor schadelijke stoffen waaraan mensen           onderzoeken verschenen over mogelijke
 Bisfenol A: schadelijk voor ogen, huid,              tijdens hun werk kunnen worden blootgesteld,      gezondheidseffecten van bisfenol A. Voor het
 luchtwegen en vruchtbaarheid                         gaat de commissie na of er uit wetenschappelijk   beoordelen van onderzoeken tot 2015 is de
 Bisfenol A (in de volksmond beter bekend als         onderzoek een concentratie is vast te stellen     commissie uitgegaan van een rapport van de
 BPA) is een weekmaker. Het wordt onder meer          waarbij geen gezondheidseffecten te               European Food Safety Authority (EFSA). Voor
 gebruikt in plastic flessen. Bisfenol A is           verwachten zijn. Deze gezondheidskundige          de onderzoeken vanaf 2015 heeft de commissie
 geclassificeerd als schadelijk voor de               advieswaarde is de basis waarop de minister       zelf gezocht naar publicaties.
 vruchtbaarheid (gevarencategorie 1B).
2        Health Council of the Netherlands | No. 2019/04                                                               2                                 4
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<pre> Samenvatting                                                                                                                    Bisphenol A | page 4 of 51
 Wel of geen monotone dosis-ffectrelatie?             Nitschke en anderen uit 1988, waarin ratten aan   voortplanting. Er zijn veel onderzoeken waarin
 Bij het afleiden van een gezondheidskundige          bisfenol A zijn blootgesteld via de luchtwegen en proefdieren (met name ratten en muizen) oraal
 advieswaarde wordt uitgegaan van het principe        waardoor ontstekingen in de neusholte             bisfenol A krijgen toegediend, waarbij wel
 dat een effect groter wordt bij een hogere dosis     ontstonden. Dit onderzoek heeft ze ook in haar    mogelijke effecten op de voortplanting zijn
 (een monotone dosis-effect relatie). Sommige         eerdere advies gebruikt en ook het advies van     bestudeerd. Veel van deze onderzoeken zijn
 onderzoekers veronderstellen dat voor bisfenol       de SCOEL uit 2013 is er op gebaseerd.             van onvoldoende kwaliteit en voldoen niet aan
 A effecten kunnen optreden volgens een niet-         Op basis van het onderzoek van Nitschke komt      de internationale richtlijnen voor
 monotone dosis-effectrelatie. De commissie is        de commissie uit op een advieswaarde van          toxiciteitsonderzoek. De commissie heeft de
 van oordeel dat daar onvoldoende bewijs voor is      3,3 mg/m3. Deze waarde is lager dan de eerdere    resultaten van het meest geschikte onderzoek,
 en hanteert daarom haar gebruikelijke                waarde van de commissie en wijkt af van het       waarin overigens geen effecten op de
 werkwijze.                                           advies van de SCOEL. Deze verschillen zijn        voortplanting werden gevonden, omgerekend tot
                                                      terug te voeren op verschillen in werkwijze. Zo   een advieswaarde in de lucht. Die waarde is
 Voorkeur voor onderzoeken naar                       heeft de commissie in het verleden geen           vergelijkbaar met de waarde die is afgeleid van
 inademing                                            onzekerheidsfactor toegepast voor de vertaling    het onderzoek van Nitschke naar bisfenol A
 Voor het afleiden van een advieswaarde in de         van diergegevens naar de mens en rondt de         blootstelling via de luchtwegen.
 lucht gebruikt de commissie bij voorkeur             SCOEL de afgeleide waarden af tot
 onderzoeken waarin blootstelling via de lucht        zogenoemde voorkeurswaarden.                      Blootstelling via de huid
 plaatsvond. Er zijn in de laatste jaren slechts                                                        De commissie concludeert op basis van de
 enkele studies uitgevoerd met blootstelling aan      Voor effecten op voortplanting ook                huidige stand van de wetenschap dat een
 bisfenol A via de lucht en daarin werden geen        onderzoeken naar inname meegenomen                huidnotatie voor bisfenol A niet nodig is.
 duidelijke nadelige effecten gevonden. Daarom        In het onderzoek van Nitschke is niet gekeken     Huidcontact draagt namelijk niet substantieel bij
 gebruikt de commissie het onderzoek van              naar mogelijke effecten van bisfenol A op de
3        Health Council of the Netherlands | No. 2019/04                                                                2                                 5
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<pre> Samenvatting                                             Bisphenol A | page 5 of 51
 aan de totale inwendige blootstelling bij een
 concentratie ter hoogte van de advieswaarde.
 Advies aan de staatssecretaris
 Voor de beroepsmatige blootstelling aan
 bisfenol komt de commissie tot een
 gezondheidskundige advieswaarde van 3,3 mg
 bisfenol A per m3 lucht. Deze waarde geldt voor
 de inhaleerbare fractie (dat deel van de in de
 lucht aanwezige stof dat kan worden ingeademd
 via mond en/of neus) en als een gemiddelde
 concentratie over een achturige werkdag.
4       Health Council of the Netherlands | No. 2019/04 2                          6
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<pre> Executive summary                                                                                                               Bisphenol A | page 6 of 51
 executive summary                                                                                      metre (m3), and an advisory value of 5 mg/m3 for
                                                                                                        respirable bisphenol A. Currently, an
                                                                                                        occupational exposure limit of 2 mg inhalable
 At the request of the Ministry of Social Affairs and irritations of the airways. Bisphenol A has been  bisphenol A per m3 air is applied. This
 Employment, the Health Council of the                restricted in several products in the EU. For     occupational exposure limit is based on a
 Netherlands has derived a health-based advisory      instance, bisphenol A is restricted in thermal    recommendation of the European Scientific
 value for bisphenol A. This advisory report has      paper since 2016 and banned from infant           Committee on Occupational Exposure Limits
 been composed by the Dutch Expert Committee          feeding bottles since 2011. Also, bisphenol A     (SCOEL) from 2013.
 on Occupational Safety (DECOS). More                 can be used in materials that are in contact with Since 2013, many studies have been published
 information on the tasks of this permanent           food, but there is a maximum amount that is       on the toxicity of bisphenol A. For the evaluation
 committee of the Health Council of the               allowed to leach out of the material.             of studies until 2015, the Committee has
 Netherlands can be found at                                                                            adopted the conclusions of the 2015 opinion of
 www.gezondheidsraad.nl. The members of the           Health-based advisory value                       the European Food Safety Authority (EFSA).
 Committee are listed on the last page of this        For hazardous substances to which people can      Publications from 2015 and onwards were
 report.                                              be occupationally exposed, the Committee          evaluated by the Committee itself.
                                                      determines whether a concentration can be
 Bisphenol A: Hazardous to eyes, skin,                derived at which no adverse health effects are    Monotonic or non-monotonic dose-
 airways and reproduction                             expected. These health-based advisory values      response relationship?
 Bisphenol A (BPA) is a plasticizer. It is used in a  are the basis at which the State Secretary can    When deriving health-based advisory values,
 wide range of consumer products such as              set an occupational exposure limit.               the principle is applied that an effect increases
 plastic bottles. Bisphenol A is classified as                                                          with an increasing dose (a monotonic dose-
 reproduction toxicant (Category 1B; for fertility).  In 1996, the Health Council has derived an        response relationship). Some investigators
 Exposure to bisphenol A can also lead to             advisory value for occupational exposure of 10    assume that for bisphenol A, effects can develop
 damage to the eyes, allergic skin reactions and      milligram (mg) inhalable bisphenol A per cubic    according to a non-monotonic exposure-
5        Health Council of the Netherlands | No. 2019/04                                                                2                                 7
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<pre> Executive summary                                                                                                               Bisphenol A | page 7 of 51
 response relationship. The Committee is of the       the SCOEL. These differences can be explained      Skin exposure
 opinion that there is insufficient evidence of such  by differences in methodology. Namely, for its     The Committee concludes that the data
 a relationship and therefore applies its usual       previous advisory value the Committee did not      available do not indicate that a skin notation for
 approach.                                            apply an uncertainty factor for extrapolation from bisphenol A is warranted. Dermal absorption
                                                      animals to humans and the SCOEL applied the        does not substantially contribute to the internal
 Preference for inhalation studies                    so-called preferred value approach.                exposure to bisphenol A, at the level of the
 For derivation of health-based advisory values in                                                       advisory value.
 air, the Committee preferentially uses studies       For effects on reproduction oral studies
 with exposures by inhalation. In recent years,       taken into account                                 Advice to the State Secretary
 only few inhalation studies with bisphenol A         In the study by Nitschke et al. (1988), possible   For occupational exposure to bisphenol A, the
 were published and in none of them, clear            effects of bisphenol A on reproduction were not    Committee derives a health-based advisory
 adverse effects have been observed. The              addressed. Many animal studies are available       value of 3.3 mg bisphenol A per m3 air. The
 Committee therefore uses the rat inhalation          (mostly with rats and mice) investigating          value relates to the inhalable fraction (the
 study by Nitschke et al. (1988), in which animals    possible effects on reproduction after oral        fraction of the substance in air that can be
 developed inflammation in the epithelium of the      exposure to bisphenol A. Many of these studies     inhaled through mouth and/or nose) and
 anterior portion of the nasal cavity. This study     are of insufficient quality and do not meet        represents a mean concentration during an 8-h
 was used by the Committee for its previous           international testing guidelines. The Committee    working day.
 report and it also forms the basis of the SCOEL      has translated the results of the most relevant
 recommendation from 2013. From the Nitschke          oral study with bisphenol A, in which no specific
 et al. (1988) study the Committee has derived a      effects on reproduction were observed, to derive
 health-based advisory value of 3.3 mg/m3. This       a value in air. This value is comparable with the
 value is lower than the Committee’s previous         advisory value that is based on the inhalation
 value, and differs from the recommendation of        study by Nitschke et al. (1988).
6        Health Council of the Netherlands | No. 2019/04                                                                2                                   8
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<pre> chapter 01 | Scope                                      Bisphenol A | page 8 of 51
 01
 scope
7      Health Council of the Netherlands | No. 2019/04 2                          9
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<pre> chapter 01 | Scope                                                                                                            Bisphenol A | page 9 of 51
 1.1 Background and objective                                               1.3 Data
 At request of the Minister of Social Affairs and Employment, the Dutch     An extensive dataset on bisphenol A toxicity is available. Also several risk
 Expert Committee on Occupational Safety (DECOS), a committee of the        assessments and opinions have been published, including those prepared
 Health Council of the Netherlands, performs scientific evaluations on the  by the EU (2003, 2008), the European Food Safety Authority (EFSA)
 toxicity of chemical substances that are used in the workplace. The        (2015), the Scientific Committee on Emerging and Newly Identified Health
 purpose of these evaluations is to recommend a health-based                Risks (SCENIHR) (2015; 2017), the Risk Assessment Committee (RAC)
 occupational exposure limit for concentrations in the air, provided the    of the European Chemicals Agency (ECHA) (2015), the Scientific
 database allows derivation of such a value. In the Netherlands, these      Committee on Occupational Exposure Limits (SCOEL) (2004, 2013) and
 recommendations serve as a basis in setting public occupational exposure   the Dutch National Institute for Public Health and the Environment (RIVM)
 limits by the Minister.                                                    (2014, 2016).1-12 Most relevant in the context of deriving a health-based
                                                                            recommended exposure limit (HBROEL) are the quantitative assessments
 In this advisory report, an evaluation is made for bisphenol A.            on health risks and exposure to bisphenol A, which are related to either
                                                                            the workplace (evaluations of the DECOS, the SCOEL, and the RAC)3,10,13
 1.2 Committee and procedure                                                or consumers (the EFSA)5,6. The recent evaluations by the RIVM are also
 This document contains the assessment of the DECOS, hereafter called       of interest, as the RIVM concluded that recently published literature
 the Committee. The members of the Committee and consulted experts are      suggests that exposure limits derived previously should be reconsidered
 listed at the end of the report.                                           (most notably the t-TDI derived by EFSA for consumers and exposure via
 In July 2017, the Committee released a draft report for public review. The food and the recommended occupational exposure limit (OEL) derived by
 Committee has taken the comments received into account, and released a     the SCOEL for workers and exposure through air).8,7
 second draft version in October 2018. No comments were received on the
 second draft report. The comments on the first draft report and the        In particular, the Committee notes the thorough evaluation of the literature
 response of the Committee can be found on the website of the Health        on bisphenol A by the EFSA.6 The Committee used this evaluation as
 Council.                                                                   starting point for the hazard assessment, although it recognizes that the
                                                                            EFSA evaluation focusses on oral exposure while inhalation is the most
8        Health Council of the Netherlands | No. 2019/04                                                             2                                  10
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<pre> chapter 01 | Scope                                                              Bisphenol A | page 10 of 51
 relevant route for the workplace.The literature reviewed by the EFSA was
 used as such and not re-evaluated by the Committee. An additional
 literature search was done until May 2018 using key words “bisphenol A”
 and “tox*” to identify literature published since the publication of the EFSA
 opinion. Relevant publications were selected based on the abstracts.
9        Health Council of the Netherlands | No. 2019/04                       2                           11
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<pre> chapter 02 | Uses, existing guidelines and standards    Bisphenol A | page 11 of 51
 02
 uses, existing
 guidelines and
 standards
10     Health Council of the Netherlands | No. 2019/04 2                           12
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<pre> chapter 02 | Uses, existing guidelines and standards                                                                                                           Bisphenol A | page 12 of 51
 2.1 Uses
                                                                                    Table 2.1. Occupational exposure limits applied word-wide. (source: Social Economic Council#; Institut
 Bisphenol A (BPA, 4,4’-Isopropylidenediphenol, CAS-number 80-05-7) is              für Arbeitsschutz der Deutschen Gesetzlichen Unfallversicherung (IFA)##)
 manufactured from phenol and acetone by an acid or alkaline catalysed                Country (organization)                 Concentration (mg/m3)              TWA        Type of OEL
                                                                                      The Netherlands                        2                                  8h         Inhalable fraction
 condensation reaction. Bisphenol A is used as a monomer in the
                                                                                      Germany (AGS)                          5                                  8h         Inhalable fraction
 manufacture of polycarbonates, which is its main use, and epoxy resins               France                                 10                                 8h
 and as an additive in plastics. It is subsequently present in a wide range of        UK                                     10                                 8h
                                                                                      EU                                     2                                  8h         Inhalable fraction
 consumer products such as plastic bottles and receipts. Due to its                   USA
 hazardous properties, bisphenol A has already been restricted in several               NIOSH                                None
                                                                                        OSHA                                 None
 products in the EU. For instance, bisphenol A is restricted in thermal paper       #
                                                                                       https://www.ser.nl/nl/thema/arbeidsomstandigheden/Grenswaarden-gevaarlijke-stoffen/Grenswaarden (accessed
                                                                                       March 3, 2019).
 since 2016 and banned from infant feeding bottles since 2011. Also, there          ##
                                                                                       Gestis Limit Values: http://limitvalue.ifa.dguv.de (accessed March 9, 2019).
 is a maximum amount of bisphenol A that is allowed to leach out of the
 materials that are in contact with food.a Other uses include for example
 flame retardants, unsaturated polyester resins and polyacrylate,                   2.3 Classification
 polyetherimide and polysulphone resins.4-6,9                                       According to the harmonised classification and labelling (ATP09) approved
                                                                                    by the EU, bisphenol A may damage fertility (Repr. 1B; H360F), causes
 2.2 Current exposure limits for the working population                             serious eye damage (Eye Dam.; H318), may cause an allergic skin
 Current occupational exposure limits of several countries are presented in         reaction (Skin sens.1; H317) and may cause respiratory irritation (STOT
 Table 2.1.                                                                         SE 3; H335).
 a
    https://chemicalsinourlife.echa.europa.eu/bisphenol-a (accessed December, 2018)
11           Health Council of the Netherlands | No. 2019/04                                                                                        2                                          13
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<pre> chapter 03 | Selection of previous evaluations and recent literature   Bisphenol A | page 13 of 51
 03
 selection of previous
 evaluations and recent
 literature
12     Health Council of the Netherlands | No. 2019/04                2                           14
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<pre> chapter 03 | Selection of previous evaluations and recent literature                                                                                          Bisphenol A | page 14 of 51
 3.1 The Netherlands Health Council (1996)                                                                the nasal cavity of rats exposed to 50 or 150 mg/m3. These effects were
 In a previous evaluation in 1996, the Committee concluded that there                                     fully reversible within 12 weeks after cessation of exposure.
 were insufficient human data available to derive a health-based                                          The Committee used a NOAEL of 10 mg/m3 as a starting point. To
 recommended occupational exposure level (HBROEL).13 Therefore, it                                        extrapolate from rat to humans, a safety factor was not considered
 derived a HBROEL for Bisphenol A based on animal data.                                                   necessary because of the absence of systemic effects and the fact that
 The Committee based its derivation on a study by Nitschke et al. (1988)a.                                the margin of safety between the NOAEL and the LOAEL for local effects
 In this study, groups of 30 rats per sex were exposed by inhalation to 0,                                is a factor 5. The Committee proposed a HBROEL of 5 mg/m3 for
 10, 50 or 150 mg/m3 bisphenol A for 6 hours/day, for 5 days/week for 13                                  respirable bisphenol A, and of 10 mg/m3 for the compound in inhalable
 weeks.14 Animals were necropsied either on the day following the last                                    form, to be averaged over an eight-hour workday (8-h TWA).
 exposure, or allowed to recover for 4 or 12 weeks. Terminal body weights
 of male rats exposed to 10, 50 or 150 mg/m3 were not different from                                      3.2 The Scientific Committee on Occupational Exposure Limits
 controls. Body weights of female rats in the highest dose group were                                            (SCOEL) (2013)
 decreased (approximately 11%). The following effects on organ weight                                     In 2013, the SCOEL published an update of its earlier criteria document
 were noted: decreased absolute liver weight in males at 10 or 150 mg/m3;                                 (2004) and amended its original recommendation.9,10 The SCOEL
 decreased absolute liver and kidney weights in females exposed to 150 mg/                                evaluated the existing toxicological literature on bisphenol A up to May
 m3; increased relative brain weights in females exposed to 50 or 150 mg/m3;                              2012, covering the data published since the publication of previous
 and increased relative lung weights in females exposed to 150 mg/m3.                                     evaluation.
 These changes were not accompanied by microscopic changes. Enlarged                                      To establish a recommended OEL, the SCOEL focused on the available
 ceca were found the day after exposure but were not apparent after 12                                    data relating to inhalation exposure. The SCOEL identified respiratory
 weeks recovery. Examination of the respiratory tract revealed very slight to                             tract irritation as the most critical effect of bisphenol A upon inhalation and
 slight epithelial hyperplasia and chronic inflammation of the submucosa in                               therefore used the same study as the Committee for derivation of an OEL,
                                                                                                          i.e. that of Nitschke et al. (1988).14 Also the SCOEL derived a NOAEL of
                                                                                                          10 mg/m3, with mild olfactory epithelium inflammation as the critical
 a
   The Committee referred to a ‘study from 1988,reported by Webb (1990)’. This appears to be the study by
   Nitschke et al. (1988), which was also used by the SCOEL.
13         Health Council of the Netherlands | No. 2019/04                                                                                             2                                 15
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<pre> chapter 03 | Selection of previous evaluations and recent literature                                                                                             Bisphenol A | page 15 of 51
 adverse effects at 50 and 150 mg/m3.14 No evidence of systemic toxicity                                      starting point, the EFSA determined in a weight of evidence approach the
 was noted in this study according to the SCOEL.                                                              likelihood that a particular effect was associated with exposure to
 This NOAEL was taken as the starting point for deriving a recommended                                        bisphenol A. Regarding general toxicity, bisphenol A was considered to
 OEL, and divided by an assessment factor of 3 to cover the uncertainties                                     affect kidney and liver weight based on observations in parental animals
 related to the inter-species extrapolation. Using the preferred value                                        and in the following generations of rats (Tyl et al., 2002)15 and mice (Tyl et
 approach, 3 mg/m3 was rounded to a recommended value of 2 mg/m3 (as                                          al., 2008)16 examined in multi-generation studies. In addition, the EFSA
 inhalable dust). The SCOEL also noted that concern has been raised for                                       indicated that bisphenol A might induce several other adverse health
 systemic toxicity after long-term exposure. This was related to the effects                                  effects at exposure levels below the NOAEL for general toxicity, involving
 on kidney weight and liver seen in rodents, for which a BMDL10 of 3.5 mg/                                    effects on the mammary gland, as well as on reproduction, metabolism,
 kg bw and a NOAEL of > 5 mg/kg bw was derived, respectively. According                                       neuro-behaviour and the immune system. However, due to methodological
 to the SCOEL, these doses are equivalent to an inhalation exposure level                                     shortcomings in the evaluated studies, these effects were not considered
 of 34 and 49 mg/m3, respectively. Because the observed liver and kidney                                      ‘likely’ by the EFSA and therefore not used as starting point for deriving a
 effects were very mild even at the highest dose levels, the recommended                                      health-based guidance value.
 value was considered sufficiently conservative by the SCOEL (covering                                        The EFSA used general toxicity observed in a reproduction toxicity study
 also the extrapolation to long-term exposure, and possible remaining inter-                                  by Tyl et al. (2008)16 as starting point to derive a TDI. In this 2-generation
 and intra-species differences in toxicokinetics and toxicodynamics).                                         study in mice, animals (28 per sex per group) received bisphenol A in the
                                                                                                              diet at concentrations of 0, 0.018, 0.18, 1.8, 30, 300, or 3500 ppm
 3.3 The European Food Safety Authority (EFSA) (2015)                                                         (equivalent to 0, 0.003, 0.03, 0.3, 5, 50, or 600 mg bisphenol A/kg bw/day.
 In 2015, the EFSA updated its opinion on the human health hazards of                                         The only systemic effects in the F0-generation were observed in males:
 bisphenol A exposure, based on an extensive assessment of the available                                      centrilobular hepatocyte hypertrophy at ≥ 300 ppm (50 mg/kg bw/day),
 literature.4-6 In this hazard evaluation, the EFSA also reconsidered its                                     and reduced body weight, increased kidney and liver weights,
 tolerable daily intakea (TDI) derived previously. For the selection of a                                     centrilobular hepatocyte hypertrophy, and renal nephropathy at ≥ 3500
                                                                                                              ppm (600 mg/kg bw/day). There were no effects on fertility reported. Only
 a
    An estimate of the amount of a substance in food or drinking water which is not added deliberately (e.g.  at the highest exposure dose, developmental effects were observed.
    contaminants) and which can be consumed over a lifetime without presenting an appreciable risk to health.
14          Health Council of the Netherlands | No. 2019/04                                                                                               2                                  16
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<pre> chapter 03 | Selection of previous evaluations and recent literature                                                            Bisphenol A | page 16 of 51
 These involved reduced F1/F2-generation weanling body weight, reduced       this BMDL10 and its corresponding HED, the EFSA applied an extra safety
 weanling spleen and testes weights (with seminiferous tubule hypoplasia),   factor of 6 in addition to a factor of 25 (consisting of a factor of 2.5 for
 slightly delayed preputial separation (PPS), and increased the incidence    interspecies differences, and a factor of 10 for intraspecies differences),
 of undescended testes (only in weanlings, which did not result in adverse   resulting in a total uncertainty factor of 150. Applying this uncertainty
 effects on adult reproductive structures or functions), and increased       factor subsequently resulted in a t-TDI of 4 μg/kg bw/day.
 gestational length by 0.3 days in F1/F2-generations. The EFSA concluded
 that alteration in kidney weight was the most critical effect because other 3.4 The Risk Assessment Committee (RAC) (2015)
 effects were only observed at higher doses. However, the EFSA noted         In 2015, the RAC of the European Chemicals Agency (ECHA) published
 that there were remaining uncertainties about possible toxic effects below  an opinion on an Annex XV dossier proposing restrictions on bisphenol A.3
 the dose at which effects on the kidney are observed. The EFSA included     This restriction proposal concerns the health risks identified for pregnant
 these effects in an overall uncertainty evaluation to derive a temporary    workers and consumers (in particular for their unborn children) exposed to
 tolerable daily intake (t-TDI) as described hereafter.6                     bisphenol A contained in thermal paper they may handle.
 The EFSA performed benchmark dose analyses on the multi-generation          The RAC took a number of evaluations into account (EU, 2003, 2008);
 reproductive toxicity study in mice (Tyl et al., 2008)16 and subsequently   SCOEL, 2014; SCENIHR, 2015, with an emphasis on the most recent
 used a BMDL10a of 8.96 mg/kg bw/day, for alteration in kidney weight in     evaluation of EFSA 2015.1,2,4-6,9-11
 mice, as a starting point for derivation of a TDI.6                         Taking into account the overall data set, the RAC agreed with the HED
 The BMDL10 based on mouse data was translated into a human dose level       approach applied by the EFSA for the risk assessment of bisphenol A. The
 (the human equivalent dose (HED)), by considering the ratio between         RAC considered that for kidney effects, the HED of approximately 600 μg/
 internal exposure (area under the curve; AUC) derived from toxicokinetic    kg bw per day would result in a DNEL of 24 μg/kg bw/day using default
 studies in mice and the internal exposure predicted using physiologically-  assessment factors (600 divided by 2.5 (remaining factor for interspecies
 based pharmacokinetic (PBPK) modeling in humans. Using AUCs of              differences) and divided by 10 (factor for intraspecies differences).
 0.244 (mice) and 3.6 nmol × h × L-1 (human reference value), the BMDL10     The RAC, however, also agreed with the EFSA that kidney effects are not
 of 8.96 mg/kg bw/day in mice was translated to an HED of 609 μg/kg bw/      the most critical effects of bisphenol A , noting that studies on other
 day. Due to the remaining uncertainties about possible toxic effects below  endpoints do not provide a sufficiently robust starting point. The EFSA
15       Health Council of the Netherlands | No. 2019/04                                                                 2                                 17
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<pre> chapter 03 | Selection of previous evaluations and recent literature                                                             Bisphenol A | page 17 of 51
 argued that other adverse effects could occur starting from a HED of 100       furthermore concluded that these effects could also be possible at 0.5 μg/
 μg/kg bw/day (approximately 6-fold lower than the HED for kidney effects).     kg bw/day, but noted that the studies supporting these conclusions have
 Applying an additional factor of 6 to a DNEL of 24 μg/kg bw/day results in     limitations and that a more detailed weight of evidence analysis of the
 a DNEL 4 μg/kg bw/day, which is equal to the t-TDI derived by the EFSA.        underlying data is needed to determine whether effects at this lower dose
 The RAC therefore supported the t-TDI derived by the EFSA for the use          level should be considered adverse.
 as an oral-DNEL for the general population. The corresponding oral DNEL        The RIVM stated that the recent insights into the immunotoxicity warrant
 for workers was therefore set at 8 μg/kg bw/day (due to a 2-fold lower         reconsideration of both the t-TDI derived by the EFSA and the DNELs
 uncertainty factor for intraspecies differences).                              derived by the RAC. The available information on occupational exposure
                                                                                and the exposure limit applied led the RIVM to conclude that there is a
 3.5 The National Institute for Public Health and the                           health risk for workers from inhalation of bisphenol A during the
       Environment (RIVM) (2014, 2016)                                          manufacture of bisphenol A, and possibly during the manufacture of epoxy
 In 2016, the RIVM published a recommendation on bisphenol A for human          resins. However, the RIVM did not address the derivation of a health-
 and environmental risk management.7 In this report, the RIVM provided an       based advisory value for occupational exposure by inhalation.
 overview of human health issues and assessments published up to March
 2014.1-11 In addition, the RIVM identified a number of scientific publications 3.6 Recent literature
 on the developmental effects of bisphenol A exposure on the immune             Literature published after finalisation of the EFSA report is summarised in
 system that were not taken into account by the EFSA and the RAC. For an        Annex B.
 evaluation of these studies, the Committee refers to a workshop report on
 developmental immunotoxicity studies with bisphenol A in the context of        Inhalation exposure
 the opinion by the EFSA.17                                                     Data on adverse effects after inhalation exposure to bisphenol A are very
 The RIVM concluded that for pre- or perinatal exposure to bisphenol A, a       limited. The only recent inhalation study is a study on the effects on the
 LOAEL of 5 μg/kg bw/day can be derived for effects on the immune               oestrous cycle, spatial learning, and memory in rats exposed by inhalation
 system, possibly resulting in increased risk of food intolerance,              to 0, 10, 30, and 90 mg/m3 for 6 hr/day, 5 days/week for 8 weeks
 inflammation and sensitivity to infections in the offspring in rats. The RIVM  (equivalent to 1.4, 4.3 and 12.9 mg/kg bw).18 The particle size mostly
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<pre> chapter 03 | Selection of previous evaluations and recent literature                                                                                      Bisphenol A | page 18 of 51
 ranged from 2.10 to 7.0 μm, accounting for 75% of the total distribution.   • A statistically significant dose-response relationship has been
 Mortality, clinical signs, body weight, haematology, serum chemistry,           established.
 oestrous cycle parameters, performance in the Morris water maze testa,      The Committee is of the opinion that none of the recently published
 and organ weights, as well as gross and microscopic histopathological       studies fulfils all criteria. Many studies have only applied a single dose of
 findings, were studied. At the highest exposure concentration, slight but   bisphenol A. In addition, for most of them it is not clear whether the
 statistically significant decreases were observed for total serum           studied effect should be considered as an adverse health effect. The
 cholesterol and adrenal gland weight. However, these effects were not       reported studies often analysed changes in very specific and
 considered adverse and there were no effects reported on the other          unconventional parameters, for which it is not clear whether they should
 endpoints in any of the male or female rats exposed to bisphenol A.         be considered as, or to lead to, adverse health effects. These effects
 The Committee derives a 90-d NOAEL of 90 mg bisphenol A/m3 from this        include, for example, changes in gene expression, protein levels, receptor
 study.                                                                      expression, DNA methylation and signalling pathways, and may equally
                                                                             well reflect reversible physiological adaptations preventing adverse
 Oral exposure                                                               outcomes.
 A large number of additional studies on bisphenol A toxicity in rats and    Several well-conducted studies have been published on relevant
 mice after oral administration has been published.19-73 Most of these       endpoints and include multiple doses. However, in these studies no
 studies do not follow international guidelines. A summary table of the      adverse effects have been observed, or only at relatively high doses.
 recent publications (from 2014 until May 2018) on effects of bisphenol A at Delclos et al. (2014) performed a FDA/GLP-compliant subchronic study
 dose levels below the starting point of EFSA’s assessment (9 mg/kg bw/      with rats exposed from gestation until start of labor, and the pups
 day) is provided in Annex B. To determine whether one or more of these      subsequently from birth to day 90.62 Clear adverse effects were only
 studies might provide an alternative starting point for deriving a HBROEL,  observed at doses of 100 and 300 mg/kg bw/d. Another well-conducted
 the Committee assessed them by applying three criteria:                     study is the core study of the CLARITY-bisphenol A programa. The results
 • More than one dose level has been tested
                                                                               Consortium Linking Academic and Regulatory Insights on BPA Toxicity: a research program developed by NIEHS,
 • The studied effect parameter is considered as a relevant adverse
                                                                             a
                                                                               NTP, and the U.S. Food and Drug Administration (FDA). This program consists of two parts: a core study
    health effect                                                              according to federal regulatory and statutory guidelines for toxicity testing and grantee studies conducted by
                                                                               university researchers but testing a broader range of health endpoints.
17       Health Council of the Netherlands | No. 2019/04                                                                                     2                                                19
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<pre> chapter 03 | Selection of previous evaluations and recent literature                                                            Bisphenol A | page 19 of 51
 of this extensive FDA-guideline perinatal and chronic extended dose-          Overall, the Committee concludes that the recent literature does not
 range finding study of bisphenol A in rats are currently only available in a  provide an alternative for the starting point used previously by the EFSA,
 draft report.72 In this study, rats were administered bisphenol A by oral     the RAC or the Committee. The Committee notes that several studies
 gavage (0-25 mg/kg) from gestation day 6 continuously, directly to pups       have been published that suggest that bisphenol A causes developmental
 from postnatal day 1 until termination at one or two years. In addition to a  effects at exposure levels far below the EFSA starting point. However, all
 continuous study, a stop-dose group was included with animals dosed           of these studies have limitations as noted by both the EFSA and the
 until post-natal day 21. In this study, no treatment-related effects were     Committee above. Importantly, in addition to limitations in design and
 observed. The other studies within the CLARITY project did not follow         reporting, these studies describe findings that are not consistent with
 international guidelines but used animals raised in the same conditions       results obtained in other studies. In view of the extensive dataset on
 and exposed to the same doses of bisphenol A as the core study. Most of       bisphenol A, the Committee considers the likelihood of incidental findings
 these studies do not report any effects57,67-69,73 or do not report on effect high. The Committee is therefore of the opinion that these studies are not
 parameters that are considered to be related to adverse health                suitable for risk assessment and no conclusions can currently be drawn
 effects19,59,63 (see Annex B).                                                based on the findings reported in these studies.
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<pre> chapter 04 | Hazard assessment                          Bisphenol A | page 20 of 51
 04
 hazard
 assessment
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<pre> chapter 04 | Hazard assessment                                                                                                    Bisphenol A | page 21 of 51
 4.1 Evaluation of (non-)monotonic dose response relationships                 The authors noted that a formal method for (dis)proving NMDRs does not
 The Committee notes that it has been suggested that bisphenol A-induced       exist. They applied a tiered approach, in which first literature was identified
 effects can develop according to a non-monotonic dose-response                that indicated NMDR for a substance relevant within the area of food
 relationship (NMDR). NMDRs have been defined as non-linear                    (other than essential nutrients, pharmaceuticals, hormones, radioactive
 relationships between dose and effect where the slope of the dose-            substances, nanomaterial or abstracts of botanicals) based on abstract
 response curve changes sign somewhere within the range of doses               and title. Thereafter, the relevant studies were first selected by applying
 examined (a ‘u-shaped’ dose-response curve). For bisphenol A, NMDRs           criteria relating to the indication of possible NMDRs, number of dose
 have been reported for various endpoints in in vitro, in vivo, and human      groups applied, substance composition, and reliability (see for details the
 studies (reviewed in Vandenberg 2014).51 The existence of such NMDRs          publication by Beausoleil et al. (2016)74).
 would conflict with the methodology used by the Committee to establish        The dose-responses described in the remaining studies were analysed
 HBROELs, which is based on monotonic dose-response relationships.             using six ‘checkpoints’ consisting of the following questions:
 Therefore, the Committee will first consider the potential role of NMDRs in   1. Can the apparent NMDR be explained by random fluctuations around a
 bisphenol A toxicity.                                                             horizontal dose-response (=no effect at all)?
                                                                               2. Can the apparent NMDR be explained by random fluctuations around a
 After the publication of the EFSA evaluation6, EFSA initiated a project in        monotonic dose response?
 which the evidence for the NMDR hypothesis was evaluated by critically        3. Can the apparent NMDR be explained by one single potential outlying
 reviewing the scientific peer-reviewed literature from 2002 onwards for           dose group?
 bisphenol A and other substances for which NMDRs have been reported.74        4. Is the effect size in one of the directions of the NMDR smaller than 5%?
 Although the focus in this project differed from the Committee’s focus (i.e.  5. Is the steepness of the dose-response curve outside the range of
 exposure via food by oral consumption versus exposure via inhalation),            biologically plausible/realistic dose-response shapes?
 the Committee considers this review of the NMDR hypothesis for                6. Does the apparent NMDR consist of more (or less) than two directions?
 bisphenol A of relevance since it addresses systemic effects and it is likely The checkpoints were applied to weigh the evidence of a potential NMDR,
 that such effects could also occur after exposure by inhalation.              without ranking them in a particular order. The total number of fulfilled
                                                                               checkpoints (i.e. when the answer to the question was ‘no’) was used to
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<pre> chapter 04 | Hazard assessment                                                                                                    Bisphenol A | page 22 of 51
 rank the datasets; datasets with more fulfilled checkpoints were              The Committee concludes that there is currently insufficient evidence to
 considered to contain more evidence for NMDR than datasets with fewer         support the existence of a NMDR for bisphenol A. Based on the weight of
 fulfilled checkpoints.                                                        evidence available, the Committee considers a NMDR for bisphenol A not
                                                                               likely and therefore, does not take it into account in the hazard
 The Committee has retrieved and examined the detailed results of the          assessment.
 review by Beausoleil et al. (2016)74 on bisphenol A. When focussing on in
 vivo studies, only 4 studies (Angle et al. (2013)75; Kendig et al. (2012)76;  4.2 Hazard identification
 Tyl et al. (2002)15 and Zsarnovszky et al. (2005)77) fulfilled the            In the evaluations published before on bisphenol A,1,3,6,7,9 data from animal
 requirements set for the initial relevance and reliability assessment. These  experiments are summarised that indicate that bisphenol A can cause a
 four studies contained 34 datasets, which were evaluated according to the     diversity of toxic effects, including acute toxicity, irritation, sensitisation,
 6 checkpoints outlined above. None of the datasets fulfilled all 6 or 5       repeated dose toxicity (including inflammation and hyperplasia of the
 checkpoints, 6 datasets fulfilled 4 checkpoints, 5 datasets fulfilled 3       airways after inhalation exposure, and effects on liver and kidneys), and
 checkpoints, 4 datasets fulfilled 2 checkpoints, 16 datasets fulfilled 1      developmental toxicity. Also with respect to specific developmental toxicity,
 checkpoint, and 3 datasets fulfilled none of the checkpoints (see Table       a diversity of effects are reported in recent studies, including effects on the
 4.1).                                                                         prostate, mammary gland, brain and behaviour, and recently, on the
                                                                               immune system.
 Importantly, only 3 datasets meet checkpoint 3, which means that for most
 datasets a possible NMDR can be explained by one single potential             4.2.1 Assessment of relevant studies
 outlying dose group. Also, the endpoints for which 3-4 checkpoints are        In previous evaluations by the EU, the SCOEL and the EFSA, the
 fulfilled show no consistency between studies (i.e. involve different types   quantitative hazard assessment is based on animal data as it is concluded
 of effect and different tissues) and for most effects analysed, it is unclear that the available human data are not suitable. The Committee adopts this
 whether it should be considered an adverse health effect.                     conclusion and will focus therefore on animal data. Although inhalation
                                                                               studies are most relevant for the occupational situation, the Committee
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<pre> chapter 04 | Hazard assessment                                                                                                                                                        Bisphenol A | page 23 of 51
 Table 4.1. Results of the analysis of Beausoleil et al. (2016) on the in vivo studies on bisphenol A that met the reliability criteria. Red: answer to the checkpoint = yes (checkpoint
 is not met); green: answer to checkpoint = no (checkpoint is met)
  First    Animal  Strain            Duration of administration of test substance (if           Outcome/effect/ endpoint          Number Checkpoint Checkpoint Checkpoint Checkpoint Checkpoin Checkpoint           Total
  author   species (EFSA: id_strain) developmental include age).                                measured (that indicates NMDR)    of dose 1 fulfilled 2 fulfilled 3 fulfilled 4 fulfilled t 5 fulfilled 6 fulfilled number of
                                     (EFSA: exp_duration and id_duration_unit)                  (EFSA: effect_desc)               levels                                                                            checkpoints
                                                                                                                                  tested                                                                            fulfilled
                                                                                                                                  (excl.
                                                                                                                                  negative
                                                                                                                                  control)
  Angle    Mouse   CD-1              GD 9-18                                                    Body weight at day of birth       5                                                                                 0
  Angle    Mouse   CD-1              GD 9-18                                                    Body weight week 3                5                                                                                 1
  Angle    Mouse   CD-1              GD 9-18                                                    Body weight week 13               5                                                                                 1
  Angle    Mouse   CD-1              GD 9-18                                                    Body weight week 19               5                                                                                 1
  Angle    Mouse   CD-1              GD 9-18                                                    Body weight average weeks 7-19    5                                                                                 1
  Angle    Mouse   CD-1              GD 9-18                                                    Energy intake week 3-4            5                                                                                 1
  Angle    Mouse   CD-1              GD 9-18                                                    Energy intake week 4-5            5                                                                                 1
  Angle    Mouse   CD-1              GD 9-18                                                    Energy intake week 15-19          5                                                                                 1
  Angle    Mouse   CD-1              GD 9-18                                                    Gonadal fat pad weight            5                                                                                 1
  Angle    Mouse   CD-1              GD 9-18                                                    Renal fat pad weight              5                                                                                 2
  Angle    Mouse   CD-1              GD 9-18                                                    Total abdominal fat               5                                                                                 1
  Angle    Mouse   CD-1              GD 9-18                                                    Cell number in gonadal fat pad    5                                                                                 4
  Angle    Mouse   CD-1              GD 9-18                                                    Gonadal adipocyte volume          5                                                                                 3
  Angle    Mouse   CD-1              GD 9-18                                                    Cell number in renal fat pad      5                                                                                 4
  Angle    Mouse   CD-1              GD 9-18                                                    Renal adipocyte volume            5                                                                                 3
  Angle    Mouse   CD-1              GD 9-18                                                    Liver weight                      5                                                                                 2
  Angle    Mouse   CD-1              GD 9-18                                                    Glucose tolerance AUC             5                                                                                 2
  Angle    Mouse   CD-1              GD 9-18                                                    Insulin tolerance AUC             5                                                                                 1
  Angle    Mouse   CD-1              GD 9-18                                                    Serum Leptin                      5                                                                                 3
  Angle    Mouse   CD-1              GD 9-18                                                    Serum Adiponectin                 5                                                                                 4
  Kendig   Mouse   CD-1              from arrival until necropsy, including pre-mating, mating, Sperm count                       5                                                                                 4
                                     pregnancy, …P and F1 generation until necropsy
  Kendig   Mouse   CD-1              from arrival until necropsy, including pre-mating, mating, Sperm motility                    5                                                                                 4
                                     pregnancy, …P and F1 generation until necropsy
  Kendig   Mouse   CD-1              on PND14 for female F1 and PND21 for male F1               Male AGD at PND 14                5                                                                                 0
  Kendig   Mouse   CD-1              on PND14 for female F1 and PND21 for male F1               Male AGD at PND 21                5                                                                                 1
  Tyl      Rat     Sprague-Dawley    3-generations (F0-F3)                                      Liver weight                      6                                                                                 1
  Tyl      Rat     Sprague-Dawley    3-generations (F0-F3)                                      Relative liver weight             6                                                                                 1
  Tyl      Rat     Sprague-Dawley    3-generations (F0-F3)                                      Relative liver weight             6                                                                                 1
  Tyl      Rat     Sprague-Dawley    3-generations (F0-F3)                                      Relative liver weight             6                                                                                 2
  Tyl      Rat     Sprague-Dawley    3-generations (F0-F3)                                      Paired testes weight              6                                                                                 1
  Tyl      Rat     Sprague-Dawley    3-generations (F0-F3)                                      Paired testes weight              6                                                                                 1
  Tyl      Rat     Sprague-Dawley    3-generations (F0-F3)                                      Anogenital distance in F2 females 6                                                                                 0
  Tyl      Rat     Sprague-Dawley    3-generations (F0-F3)                                      Absolute liver weight             6                                                                                 4
  Tyl      Rat     Sprague-Dawley    3-generations (F0-F3)                                      Relative liver weight             6                                                                                 3
  Zsarnovs Rat     Sprague-Dawley    n.a.                                                       pERK-IR cell number in the        7                                                                                 3
  zky                                                                                           cerebellar cortex at P10.
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<pre> chapter 04 | Hazard assessment                                                                                                                   Bisphenol A | page 24 of 51
 also considers systemic effects observed in oral studies relevant when      (1988)14, from which a NOAEL of 10 mg/m3 was established, the critical
 these are also expected to occur after inhalation exposure.                 study for quantitative hazard assessment. This NOAEL is based on the
 For the previously proposed occupational exposure limits or tolerable       absence of inflammation of the upper respiratory tract epithelium and
 intake level, one of two studies has been used as starting point. The       olfactory epithelium, and relates to the inhalable fraction. This study was
 DECOS, the SCOEL and the EU have used the repeated dose inhalation          previously used by the Committee to derive an HBROEL of 10 mg/m3.13
 study in rats by Nitschke et al. (1988)14, whereas the EFSA has used the    According to its current guideline, the Committee applies an uncertainty
 oral multi-generation studies in mice by Tyl et al. (2002, 2008)15,16 The   factor of 3 for intraspecies extrapolation, which results in a HBROEL of:
 Committee considers all these guideline-compliant studies suitable for risk
 assessment. Tyl et al. (2002, 2008)15,16, however, applied the oral route   10 mg/m3 / 3 = 3.3 mg/m3 (inhalable fraction)
 which is considered less suitable to derive an advisory value for
 concentrations in air.                                                      This approach of deriving an advisory value is similar to the approach
 As outlined in section 3.6 (Recent literature), the Committee concludes     applied by the SCOELa.
 that the recent low dose studies on bisphenol A do not provide an
 alternative starting point to derive an HBROEL.                             The Committee notes that the inhalation studies do not address potential
                                                                             effects on the offspring. Three oral repeated dose toxicity studies that are
 4.3 Quantitative hazard assessment                                          suitable for risk assessment have included potential developmental
 In line with the earlier reports of the DECOS, the SCOEL and the EU, the    effects. In the recent perinatal and chronic toxicity study in rats by the
 Committee prefers data on inhalation exposure as starting point for         NTP, no treatment-related findings were reported up to the highest dose
 deriving a HBROEL, since inhalation exposure is most relevant for           tested (25 mg/kg bw/d). Also, in the multi-generation studies of Tyl et al.
 occupational settings. Only two inhalation studies are available: the       (2002, 2008) in rats and mice no developmental effects were noted. The
 13-week repeated dose inhalation study of Nitschke et al. (1988)14 and the  most critical effect observed consisted of alterations in kidney weight in
 recently published 8-week repeated dose inhalation study by Chung et al.    mice, as has been outlined by EFSA.6 Based on a corresponding BMDL10
 (2017)18 Since Chung et al. (2017).18 did not report any adverse effects up
 to 90 mg/m3, the Committee considers the study of Nitschke et al.           a
                                                                               Because the SCOEL applies the preferred value approach, it recommends a limit value of 2 mg/m3.
23       Health Council of the Netherlands | No. 2019/04                                                                               2                                       25
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<pre> chapter 04 | Hazard assessment                                                                                                                              Bisphenol A | page 25 of 51
 of 8.96 mg/kg bw/day, an HBROEL can be derived taking into account                                      4.4 Skin notation
 default uncertainty and conversion factors:                                                             The Committee applies the ECETOC strategy to decide on assigning a
                                                                                                         skin notation.78 In this strategy, a skin notation is warranted when
 8.96 mg / (3 x 3 x 2a) = 500 µg/kg bw/d                                                                 absorption through 2000 cm2 exposed skin in 1 h is estimated to exceed
                                                                                                         10% of the systemic dose after inhalation during a day exposure at a level
 500 x 70/10b = 3.5 mg/m3                                                                                of the HBROEL (if HBROEL is based on a systemic toxicity endpoint). This
                                                                                                         is the case when the Critical Absorption Value (CAV; the rate of absorption
 This value, based on oral toxicity studies that take effects on the offspring                           above which dermal exposure is considered an important contributor to
 into account, is comparable with the HBROEL based on a repeated dose                                    the total exposure) exceeds:
 inhalation toxicity study. Thus, it is assumed that the HBROEL proposed
 also covers these endpoints. The Committee notes that a derivation based                                (10 [m3] x OEL [mg/m3] x f x 0.1)/2,000 [cm2]
 on oral studies results in a substantially higher value than the t-TDI
 derived by the EFSA. This difference is explained by the fact that the                                  in which 10 m3 is the human inhalation volume per 8h working day, f is the
 Committee does not take into account potential toxicokinetic differences                                absorption factor for inhalation (here assumed to be 1), 0.1 denotes the
 between species, and applies a less conservative approach for its                                       10% criterion, 2,000 cm2 is the surface area of the hands and forearms,
 uncertainty factors.                                                                                    and OEL is in this case the HBROEL. Thus the CAV will be:
 The Committee is aware of the fact that in some studies,                                                (10 [m3] x 3.3 [mg/m3] x 1 x 0.1)/2,000 [cm2] = 1.65 μg/cm2*h
 neurodevelopmental effects after oral administration have been reported
 at doses in the range of 0.5 µg bisphenol A/kg bw. However, as outlined                                 The RAC3 notes a publication by Marquet et al. (2011)79, reporting the
 above, the Committee considers these studies not suitable for deriving an                               measured cutaneous absorption flow of bisphenol A, varying between
 health-based advisory value.                                                                            0.026 μg.cm-2.h-1 (minimum) and 0.331 μg.cm-2.h-1 (maximum). Because
                                                                                                         the estimated CAV is higher than this reported cutaneous absorption flow
 a
   3 for intraspecies differences, 3 for interspecies species, 2 for subchronic to chronic extrapolation range, the Committee considers a skin notation not necessary.
 b
   70 kg for assumed body weight, 10 m3 for assumed inhalation volume in workers
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<pre> chapter 04 | Hazard assessment                                               Bisphenol A | page 26 of 51
 4.5 Groups with increased risk
 Bisphenol A is classified as a substance toxic for reproduction (Cat 1B).
 Furthermore, some recent animal data suggest that developmental effects
 can occur at relatively low exposure levels. Although the Committee
 considers these studies not suitable for risk assessment, it is of the
 opinion that pregnant women and their offspring represent groups at
 increased risk.
 4.6 Conclusions and recommendation
 For bisphenol A, the Committee recommends an occupational exposure
 limit of 3.3 mg/m3 (inhalable fraction), as a mean concentration during an
 8-h working day. In addition, the Committee concludes that a skin notation
 is not indicated.
25        Health Council of the Netherlands | No. 2019/04                   2                           27
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<pre> Literature                                               Bisphenol A | page 27 of 51
 literature
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<pre> Literature                                                                                                                     Bisphenol A | page 28 of 51
 1.
    European Union. European Union Risk Assessment Report                       7
                                                                                   RIVM. National Institute for Public Health and the Environment.
    4,4’-isopropylidenediphenol (bisphenol-A). European Commission,                Bisphenol A: Part 2. Recommendations for risk management. RIVM
    Joint Research Centre 2003.                                                    Report 2015-0192: 2016.
 2.
    European Union. European Union Risk Assessment Report,                      8
                                                                                   RIVM. National Institute for Public Health and the Environment.
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<pre> Annex                                                   Bisphenol A | page 35 of 51
 annex
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<pre> Annex                                                                                                                                                             Bisphenol A | page 36 of 51
 A          recent literature
 Reference               Analysed        Study design                        Tested doses       Critical effect(s) observed Studied        Lowest effect level         Comments/Main limitations
                         species and sex                                                                                     generation                                for risk assessment
 Arambula et al.         Rat, male and   Part of CLARITY.                    0, 2.5, 25, 250,   Sex-specific transcriptional F1            2.5 µg/kg bw/d (in          Not clear whether effects
 (2016)19                female          Pregnant female rats were exposed 2,500, and 25,000    changes in mainly                          females; males no           should be considered adverse;
                                         by gavage GD6 until birth. At PND1 µg/kg bw            hypothalamus; ER                           statistical significant     no dose-response relationship
                                         the hippocampal and hypothalamic                       expression.                                Effects observed)           observed.
                                         transcriptome was analysed.
 Arambula et al.         Rat, males and  Part of CLARITY.                    0, 2.5, 25, 250,   Changes in gene              F1            No significant/treatment    Not clear whether effects
 (2017)63                females         Animals were dosed by oral gavage 2,500 and 25,000     expression pathways in the                 related effects were        should be considered adverse;
                                         continuously and were euthanized    µg/kg bw/d         amygdale (estrogen,                        observed                    no functional parameters
                                         on postnatal day (PND) 1.                              oxytocin, and vasopressin                                              assessed; limited dose-
                                         Subsequently, amygdalae were                           signalling pathways).                                                  response relationships
                                         microdissected and gene                                                                                                       observed.
                                         expression was assessed.
 Berger et al. (2016)21  Mouse, female   Mice were exposed by gavage,        0, 0.5, 20, and 50 No transgenerational effects F1, F2, F3    0.5 µg/kg bw/d              Not clear whether effects
                                         daily from GD11-birth. Ovaries were µg/kg bw           on germ cell nest                                                      should be considered adverse;
                                         collected at PND 4 and 21 from the                     breakdown and gene                                                     critical effect size unclear; no
                                         F1–F3 generations and subjected                        expression on PND 4, but it                                            apparent dose-response in
                                         to histological evaluation (trans-                     caused transgenerational                                               changes in follicle type.
                                         generational effects on germ cell                      changes in expression in
                                         breakdown; only in F2 en F3),                          multiple genes on PND 21.
                                         anti-oxidant gene expression, and
                                         distribution of follicle types.
 Borman et al. (2017)22 Mouse, female    Mice were subcutaneously exposed 0, 3 and 4 mg/kg bw   Reduction in proportion      F0            3 mg/kg bw/d                Not clear whether effects
                                         at GD1-4, uterine histo-morphology                     positive for Cad-11. No                                                should be considered adverse;
                                         and immunohistochemistry was                           statistically significant                  (effect on implantation     effect shown only at one dose;
                                         performed at GD6.                                      effects on development                     sites not stat. sig)        exposure route not relevant.
                                                                                                implantation sites.
 Brouard et al. (2016)23 Rat, male       Animals were exposed from day 15    0 and 50 µg/kg bw  Increased testis weight;     No study on   50 µg/kg bw/d               Promoting effects on
                                         30 post-partum. At day 30, testis                      increase testicular markers  developmental                             spermatogenesis not in line
                                         was weighed, histologically                            (proteins and gene           effects.                                  with most other publications;
                                         analysed at day 30. Also DNA                           expression); decrease in                                               adversity of endpoints unclear;
                                         fragmentation was determined and                       gene expression of blood-                                              only one dose tested.
                                         RNA was extracted.                                     testis-barrier.
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<pre> Annex                                                                                                                                                             Bisphenol A | page 37 of 51
  Reference             Analysed         Study design                         Tested doses         Critical effect(s) observed Studied         Lowest effect level     Comments/Main limitations
                        species and sex                                                                                          generation                            for risk assessment
 Castro et al. (2018)71 Rat, male,       BPA was administered BPA             2.4 and 10 μg/kg bw  Significantly decreased       F1            2.4 μg/kg               Only 2 doses tested; not clear
                        juvenile         subcutaneously from gestational                           mRNA and protein levels of                                          whether effects should be
                                         day 12 to parturition, pups received                      5α-R2 in prostate tissue.                                           considered adverse;
                                         the same dose from PND1-21.                               Also decreased levels of                                            sub-cutaneous exposure
                                         Thereafter, levels of 5α-R isozymes                       testosterone and                                                    route.
                                         and aromatas in prostate and                              di-hydrotestosterone, and
                                         plasma testosterone and                                   increased levels estradiol in
                                         di-hydrotestosterone were                                 plasma were measured.
                                         measured in offspring.
 Chen et al. (2016)24   Rat, adult, male Animals were exposed to 50 µg/kg 0 and 50 µg/kg bw        Decreased protein/lysine      No study on   50 µg/kg bw/d           not clear whether effects
                                         bw/d in diet for 35 weeks.                                acetylation levels and        developmental                         should be considered adverse;
                                         Thereafter testes and epididymis                          decreased histone             effects.                              only one dose tested.
                                         were dissected and weighed, testes                        acetylation in testes;
                                         and epididymis coefficients were                          increased protein
                                         calculated and one testis from each                       expression of deacetylase
                                         animal was used for protein                               Sirt1 and reduced binding
                                         determination and the other testis                        of Sirt1, together with
                                         was used HE staining and TUNEL                            increased binding of
                                         experiments.                                              oestrogen receptor β (ERβ)
                                                                                                   to caveolin-1 (Cav-1).
 Chianese et al.        Rat, male        Dams received the treatment all      0.1 mg/L in drinking Reduced the body weight       F1            130 μg/kg bw            Only one dose tested; no
 (2018)70                                over lactation and at weaning; each water (equivalent to  gain in male offspring at 45                                        guideline followed.
                                         newborn received the same            130 μg/kg bw)        postnatal days and the first
                                         treatment of the mother via drinking                      round of spermatogenesis,
                                         water. Possible effects of BPA on                         with impairment of blood
                                         the first round of spermatogenesis                        testis barrier, reactive
                                         (apoptosis, oxidative stress,                             oxygen species production,
                                         metabolism and energy                                     DNA damage and
                                         homeostasis), the male newborns                           decreased expression of
                                         were sacrificed at 17 PND, 45 PND,                        SIRT12.
                                         or 60 PND.
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<pre> Annex                                                                                                                                                          Bisphenol A | page 38 of 51
 Reference               Analysed        Study design                         Tested doses         Critical effect(s) observed Studied      Lowest effect level     Comments/Main limitations
                         species and sex                                                                                         generation                         for risk assessment
 Chung et al. (2017)18   Rats, males and Rats were exposed 6 hr/day, 5        0, 10, 30, and 90    No adverse effects                       No (significant)        Results water maze test not
                         females         days/week for 8 weeks via whole-     mg/m3                observed.                                treatment - related     reported.
                                         body inhalation. Mortality, clinical                                                               effects were observed.
                                         signs, body weight, haematology,
                                         serum chemistry, oestrous cycle
                                         parameters, performance in the
                                         Morris water maze test, and organ
                                         weights, as well as gross and
                                         histopathological findings, were
                                         assessed.
 Delclos et al. (2014)62 Rat, males and  Rat dams were dosed daily from       2.5, 8, 25, 80, 260, Clear adverse effects         F1         100,000 μg/kg bw        Conducted in compliance with
                         females         gestation day 6 until the start of   840, 2,700, 100,000, including depressed                                              FDA GLP regulations.
                                         labor, and their pups were directly  300,000 µg/kg bw/d gestational and postnatal
                                         dosed from PND1 through PND90.                            body weight gain, effects on
                                         Analyses including gestation and                          the ovary (increased cystic
                                         litter endpoints, pup preweaning                          follicles, depleted corpora
                                         survival, growth, clinical chemistry                      lutea, and antral follicles),
                                         (serum hormones), organ weights                           and serum hormones
                                         and histopathology, sperm                                 (increased serum estradiol
                                         parameters.                                               and prolactin and
                                                                                                   decreased progesterone).
 Dere et al. (2018)69    Rat, male       Part of CLARITY. Animals were        0, 2.5, 25, 250,     -                             F1         No exposure related     Extensive dose range applied.
                                         gavaged from gestational day (GD) 2,500 and 25,000                                                 effects were observed.
                                         6 until parturition, and their pups  µg/kg bw/d
                                         were directly gavaged daily from
                                         postnatal day (PND) 1 to 90 with
                                         BPA where after the testes were
                                         histologically evaluated for altered
                                         germ cell apoptosis, sperm
                                         production, and altered spermiation.
                                         DNA and RNA was isolated from
                                         isolated sperm to assess changes.
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<pre> Annex                                                                                                                                                       Bisphenol A | page 39 of 51
 Reference              Analysed        Study design                        Tested doses         Critical effect(s) observed Studied    Lowest effect level      Comments/Main limitations
                        species and sex                                                                                      generation                          for risk assessment
 Ferguson et al.        Rat, males and  Pregnant rats were dosed orally on 2.5 or 25.0 µg/kg     BPA treatment did not alter F1         No significant/treatment Only two doses tested.
 (2014)61               females         GD6–21. Body weight, pubertal       bw/d                 any measured endpoint.                 related effects were
                                        age, oestrous cyclicity, and adult                                                              observed.
                                        serum hormone levels were
                                        measured. Adolescent play, running
                                        wheel activity, flavoured solution
                                        intake, female sex behaviour, and
                                        manually elicited lordosis were
                                        assessed.
 Fergusonet al.         Rat, males and  Pregnant rats were doses on         2.5 or 25.0 µg/kg    BPA treatment did not alter F1         No significant/treatment Only two doses tested.
 (2015)60               females         GD6–21. Beginning on PND1 and       bw/d                 any measured endpoint.                 related effects were
                                        continuing until PND21, the pups                                                                observed.
                                        were orally treated with the same
                                        dose their dam had received. Body
                                        weight, pubertal age, oestrous
                                        cyclicity, and adult serum hormone
                                        levels were measured. Adolescent
                                        play, running wheel activity,
                                        flavoured solution intake, female
                                        sex behaviour, and manually
                                        elicited lordosis were assessed.
 Galyon et al. (2016)25 Rat, male and   Rat dams were fed BPA in drinking   0 and 239 µg/d       Male but not female         F1         239 µg/d during          Not clear whether effects
                        female          water from 2 weeks prior to mating  during pregnancy;    offspring had impaired                 pregnancy; 466 µg/d      should be considered adverse;
                                        and through pregnancy and           466 µg/d during      glucose tolerance at 6                 during lactation         only one dose tested.
                                        lactation. Glucose tolerance was    lactation            weeks and 6 months. Sex
                                        tested at 6 weeks and 6 months,                          and tissue-specific effects            (around 2 mg/kg bw
                                        liver and skeletal muscles tissue                        on insulin signalling                  assuming bw 125-500
                                        was collected from 3 w and 10 m                          proteins were reported.                mg).
                                        old offspring.
 Gear et al. (2017)54   Rat, male and   Part of CLARITY. Dams were dosed 0, 2.5, 25, 250,        Decreased collagen in       F1         2.5 µg/d                 High level cardiomyopathy in
                        female          with BPA from GD6 – PND0. Pups      2,500 or 25,000 µg/d hearts females at PND90                                         controls; No apparent dose-
                                        were dosed from PND1                                     and PND180;                            Females                  response; effects at PND21
                                        continuously or until PND21, to day                      cardiomyopathy incidence                                        but not statistically significant
                                        of sacrifice at PND21, PND90 or 6                        and severity increased in                                       at later time points.
                                        months. Subsequently, isolated                           females at PND21 with
                                        hearts were analysed by                                  myocardial degeneration in
                                        quantitative morphometry and                             both males and females at
                                        histopathology.                                          PND21 and PND90.
38        Health Council of the Netherlands | No. 2019/04                                                                                          2                                               40
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<pre> Annex                                                                                                                                                               Bisphenol A | page 40 of 51
 Reference              Analysed        Study design                          Tested doses           Critical effect(s) observed Studied         Lowest effect level     Comments/Main limitations
                        species and sex                                                                                            generation                            for risk assessment
 Gear and Belcher       Mouse, male and Dams were treated via the diet from   0, 0.03, 0.3, 3.0, 30, No effects on gestation       F1            4 µg/d                  Not clear whether effects
 (2017)55               female          conception, offspring were            300 ppm (doses         period, pups per litter and                                         should be considered adverse;
                                        separated from the mothers at         from 4-40,000 µg/kg    sex ratio. Increase spleen                                          no clear dose-response.
                                        PND21 and exposed via the diet        bw/d)                  weight; increase histological
                                        until 12-14 weeks of age where                               and microstructural
                                        after histological analysis was                              changes in the spleen.
                                        performed.
 Hass et al. (2016) 26  Rat, male and   Dams were gavaged BPA daily from 0, 25, and 250 µg/kg         Males: Decreased sperm       F1            -                       Effects very limited in
                        female          GD7 to PND22, analysed during life bw, 5 and 50 mg/kg        count only at lowest dose.                                          magnitude; no dose-responses
                                        (3 m-14 m). In the offspring, growth, bw                     Females: increased body                                             observed.
                                        sexual maturation, weights and                               weight late in life and
                                        histopathology of reproductive                               altered spatial learning.
                                        organs, oestrus cyclicity and sperm
                                        counts were assessed. Neuro-
                                        behavioural development was
                                        investigated using a behavioural
                                        testing battery including tests for
                                        motor activity, sweet preference,
                                        anxiety and spatial learning.
 Jardim et al. (2017)56 Mouse, male and Mice were gavaged BPA from            0 and 5 mg/kg bw/d     Impaired object recognition No study on     5 mg/kg bw/d            Only one dose tested.
                        female          PND21 to 60. The mice performed                              memory in both sexes;         developmental
                                        the behavioural memory tests and                             Impaired spatial memory in effects.
                                        the [3H] glutamate uptake and                                females and impaired
                                        NMDA receptor subunits (2A and                               passive avoidance memory
                                        2B) analyses were carried out in                             in males. Also a decrease in
                                        the hippocampus and cerebral                                 the [3H] glutamate uptake
                                        cortex of mice.                                              and NMDA receptor subunit
                                                                                                     levels in the cortical and
                                                                                                     hippocampal regions was
                                                                                                     observed depending on the
                                                                                                     sex.
 Jedeon et al. (2016)29 Rat, male and   Female rats were exposed by           0 and 5 µg/kg bw       A moderate phenotype          F1            5 µg/kg bw/d            not clear whether effects
                        female          gavage from GD1 until weaning;                               enamel hypo-mineralisation                                          should be considered adverse;
                                        pups were subsequently exposed                               was observed with 12.5% of                                          only one dose tested.
                                        at the same dose in the drinking                             enamel breakdown.
                                        water during the next 44 days (the
                                        time necessary for the full growth of
                                        the rat incisor). At PND65 enamel
                                        defects were assessed.
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<pre> Annex                                                                                                                                                              Bisphenol A | page 41 of 51
 Reference              Analysed        Study design                          Tested doses          Critical effect(s) observed Studied        Lowest effect level      Comments/Main limitations
                        species and sex                                                                                          generation                             for risk assessment
 Jiang et al. (2016)30  Rat, male       Adult male rats were administered     0.5 or 5 mg/kg bw     Reduced sperm motility,      No study on   0.5 mg/kg bw/d           Only two doses tested; very
                                        BPA for 8 weeks by gavage.                                  deformity ratios and         developmental                          limited dose-dependency.
                                        Thereafter, the reproductive system                         histological changes.        effects.
                                        was dissected.
 Jinpeng et al. 2018    Rat, male and   Part of CLARITY. Animals were         0, 2.5, 25, 250,      Most, if not all, of these   F1            No exposure related      Extensive dose range applied.
                        female          dosed by oral gavage continuously 2,500 and 25,000          alterations were found to be               effects were observed.
                                        and were euthanized on postnatal      µg/kg bw/d            transient with no persistent
                                        day (PND) 21, 90, 6 month and 1                             trend over the one-year
                                        year. At these time points, leukocyte                       period.
                                        compositions (including B cells, T
                                        cells, NK cells, granulocytes,
                                        monocytes, macrophages and
                                        dendritic cells) were determined in
                                        spleen and thymus.
 Johnson et al.         Rat, males and  Part of CLARITY.                      0, 2.5, 25, and       At-2500: more incorrect      F1            No significant/treatment No dose-response, not clear
 (2015)59               females         Animals were dosed from               25,000 µg/kg bw/d     holes sniffed on day 7                     related effects were     whether effects should be
                                        gestational day 6 to parturition, and                       -2500: females were less                   observed.                considered adverse.
                                        offspring were directly orally dosed                        likely than control females
                                        until weaning PND21. At adulthood,                          to locate the escape box
                                        animals were tested for seven days                          (latency)
                                        in the Barnes maze. Also, serum                             2.5: prolonged latency in
                                        testosterone concentrations were                            females, not significant
                                        measured.                                                   2.5: improved latency in
                                                                                                    males, significance
                                                                                                    uncertain.
                                                                                                    No differences in serum T in
                                                                                                    males and females.
 Jones et al. (2016)31  Rat, male       (1) Juvenile rats were exposed by     0, 5, 50, 500, and    No effect on neural survival F0, F1        5 µg kg/bw d             not clear whether effects
                                        gavage pre- and post-natally          5,000 µg/kg bw        or soma size in experiment                                          should be considered adverse.
                                        (GD7-PND14) and the number and                              (1); decrease in soma size
                                        size of motor neurons were                                  in retrodorsolateral nucleus
                                        examined in adulthood;                                      (RDLN) pool in experiment
                                        (2) adult rats were doses BPA for                           (2).
                                        28 days in drinking water after
                                        which the soma size of motor
                                        neurons was measured.
 Kazemi et al. (2016)32 Rat, male       Rats were exposed by gavage for       0, 5, 25, and 125 μg/ Reduced bw; seminiferous     No study on   5 µg/kg bw/d             No dose-response
                                        35 days; plasma hormone levels        kg bw                 tuble diameter and           developmental                          relationships observed.
                                        and testis were analysed at D36.                            thickness of seminiferous    effects.
                                                                                                    epithelial was decreased.
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<pre> Annex                                                                                                                                                        Bisphenol A | page 42 of 51
 Reference              Analysed        Study design                         Tested doses        Critical effect(s) observed Studied      Lowest effect level     Comments/Main limitations
                        species and sex                                                                                        generation                         for risk assessment
 Kazemi et al. (2017)66 Rat, male       BPA was administered by gavage       5, 25 and 125 µg/kg Decreased bodyweight at       F0         5 μg/kg bw              Not clear whether effects
                                        for 35 consecutive day. Thereafter,                      25 and 125 mg/kg. ALP and                                        should be considered adverse,
                                        levels of aspartate                                      AST decreased significantly                                      limited dose-response
                                        aminotransferase, alanine                                at all doses, decreased                                          relationship observed.
                                        aminotransferase and alkaline                            Beta-2 protein and
                                        phosphatase were determined, liver                       increased Gama protein
                                        histology was performed and serum                        serum levels in rats.
                                        protein were visualised.
 Ke et al. (2016)33     Mouse, male     Female mice were exposed from        0, 0.5 µg/kg bw/d   Hepatic accumulation of       F1         0.5 µg/kg bw/d          Not clear whether effects
                                        the first day of childbirth and                          triglycerides and                                                should be considered adverse;
                                        throughout lactation. After weaning,                     cholesterol; abnormalities in                                    only one dose tested.
                                        the male offspring from the mothers                      liver cells (gene expression,
                                        were daily administered BPA in                           methylation).
                                        water similar to the BPA water that
                                        their mothers drank. Part of male
                                        offspring was sacrificed at the age
                                        of 8 weeks, the rest mice were
                                        killed at the age of 10 months.
                                        Blood, liver and the perigonadal
                                        white adipose tissue were
                                        analysed.
 Komada et al.          Mouse, newborn  Exposure by oral gavage from GD6 0, 20 and 200 µg/kg     abnormal neuronal             F1         20 µg/kg bw/d           Not clear whether effects
 (2014)34                               to GD18, 20 and 200 µg/kg bw/d, At bw/d                  development (at 20 and 200                                       should be considered adverse;
                                        1 and 3 d after birth, tests for the                     µg/kg bw) and behaviour (at                                      no dose-response relationship
                                        presence of behavioural                                  200 µg/kg bw.)                                                   present.
                                        abnormalities and a detailed
                                        histologic analysis of the neocortex
                                        and were performed, respectively.
 Li et al. (2016)36     Mouse, adult,   An experimentally induced delayed    0, 60 and 600 µg/kg Improper endometrial          F0         60 µg/kg bw/d           Limited and inconsistent data
                        female          implantation model was used to       bw/d                epithelial and stromal                                           on dose-response;
                                        study the implantation process.                          functions, reduced number                                        unconventional study design
                                        Mice were exposed orally from                            of implantation sites.                                           (experimentally induced
                                        PND22, for 5 weeks, during mating                        Expression of progesterone                                       delayed implantation
                                        period until D9 of pregnancy, when                       receptor and its                                                 Model; 3x-d dosing; depletion
                                        uterine tissues were examined.                           downstream target gene                                           oestrogen from other sources).
                                                                                                 was markedly suppressed.
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<pre> Annex                                                                                                                                                             Bisphenol A | page 43 of 51
 Reference            Analysed        Study design                         Tested doses         Critical effect(s) observed Studied         Lowest effect level         Comments/Main limitations
                      species and sex                                                                                         generation                                for risk assessment
 Li et al. (2016)35   Mouse, male     Mice were treated subcutaneously     0 and 100 µg/kg bw   Decrease of insulin           No study on   100 µg /kg/d                Exposure route not relevant;
                                      for 30 days, where after the insulin                      sensitivity, GLUT1, 3 protein developmental                             not clear whether effects
                                      signalling and glucose transporters                       levels, hyper-activation of   effects.                                  should be considered adverse;
                                      in the hippocampus and prefrontal                         IR/IRS/AKT/GSK3 axis in                                                 only one dose tested.
                                      cortex were detected by western                           mouse brain.
                                      blot.
 Li et al. (2018)67   Rat, male and   Part of CLARITY. Animals were        0, 2.5, 25, 250,     No dose dependent,            F1            No exposure related         Extensive dose range applied.
                      female          dosed by oral gavage continuously    2,500 and 25,000     statistically significant                   effects were observed.
                                      and were euthanized on PND21,        µg/kg bw/d           effects observed.
                                      90, 6 month and 1 year.
                                      Subsequently, measurements of
                                      lympho-proliferation in response to
                                      mitogenic stimuli, immunoglobulin
                                      production by B cells, and cellular
                                      activation of T cells, NK cells,
                                      monocytes, granulocytes,
                                      macrophages and dendritic cells.
 Ling et al. (2016)37 Mouse, male and Pregnant mice were exposed by        0, 40 and 400 µg/kg  Neural migration in the       F1 (embryo)   40 µg/kg bw/d               Adversity of endpoint unclear;
                      female          osmotic pump from E14.5-E18.5;       bw                   cortical plate was                                                      exposure route not relevant;
                                      neuronal migration in the foetus                          significantly decreased in                                              no dose-dependency
                                      visualised at E18.5 after                                 the 40 µg/kg group (not in                                              observed.
                                      transfecting a plasmid by in utero                        the 400 µg/kg group).
                                      electroporation.
 Luo et al. (2016)38  Mouse, male and Pregnant mice were exposed via       0, 10, 100 or 1,000  Dose-dependent Increase       F1            Around 5 µg/kg bw/d         Only intermediate parameters
                      female          drinking water from GD0-PND 21.      nM in drinking water in Th17 cells in spleen                     (increase 0.5 µg/kg bw/d were included; functional
                                      At PNDs 21 and 42, blood was         (equivalent doses    (gender specific; most                      not yet stat. significant). consequences unclear.
                                      collected from the offspring for     0.5, 5 and 50 µg/kg  pronounced in females),
                                      cytokine measurement and spleens     bw, recalculated     effect on gene expression
                                      were collected for determination of  using TGD,           and cytokines.
                                      Th17 cell frequency and expression   assuming bw of
                                      of the transcription factor retinoic 30 gr).
                                      acid-related orphan receptor.
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<pre> Annex                                                                                                                                                          Bisphenol A | page 44 of 51
 Reference              Analysed        Study design                         Tested doses        Critical effect(s) observed Studied      Lowest effect level       Comments/Main limitations
                        species and sex                                                                                        generation                           for risk assessment
 Mahalingam et al.      Mice, female    Daily oral dosing of the dams in the 0.5, 20, and 50 µg/ Decreased preantral follicle F1 and F2   No significant/treatment  Not clear whether effects
 (2017)65                               respective treatment groups was      kg/day              numbers in F1. Estradiol                 related effects were      should be considered adverse,
                                        started on GD 11 and continued                           levels were decreased at                 observed.                 effect size limited, functional
                                        until the birth of pups. Some of the                     three months in F1 at 20                                           reproductive parameters not
                                        adult F1 females were used to                            and 50, µg /kg/day. In F2, at                                      reported, limited dose-
                                        generate the F2 progeny, and at                          12 m, BPA at 20 µg/kg/day                                          response relationship
                                        least one F2 female from each litter                     significantly reduced                                              observed.
                                        was euthanized at three months                           testosterone levels during
                                        and 12 months of age. Ovaries                            diestrus. In both F1 and F2,
                                        were collected or assessment of                          various ovarian mRNA
                                        histological analysis of follicle                        levels of steroidogenic
                                        numbers and health as well as                            enzymes were decreased.
                                        mRNA levels of steroidogenic
                                        enzymes.
 Mandrup et al.         Rat, male and   Dams were gavaged BPA daily from 0, 0.025, 0.25, 5,      Male offspring showed         F1         No consistent effect.     No dose-response
 (2016)39               female          GD7 to the day before expected       and 50 mg/kg bw     increased mammary                        Increased mammary         relationship, inconsistent
                                        birth. Effects on the mammary                            outgrowth on pup day                     outgrowth was observed results.
                                        gland in the offspring were                              PND22; increased                         only in males, only at a
                                        measured at 22, 100, and 400 days                        prevalence of intraductal                dose of 0.025 µg/kg
                                        of age.                                                  hyperplasia was observed                 bw/d). Intraductal
                                                                                                 in females at PD 400,                    hyperplasia was
                                                                                                                                          observed in females,
                                                                                                                                          only at a dose of 0.25
                                                                                                                                          µg/kg bw/d).
 Medwid et al. (2016)40 Mouse, adult,   Pregnant mice fed 25 mg/kg diet      0 and 25 mg kg food Increased adrenal gland       F1         4.8 mg/kg bw/d            Not clear whether effects
                        male            from GD7 until delivery. At 8 weeks  pellet              weight (males and females);                                        should be considered adverse;
                                        of age, offsprings were sacrificed,                      elevated plasma corticos-                (recalculated using       critical effect size unclear; only
                                        blood samples and adrenalglands                          terone levels (males and                 TGD, assuming bw of       one dose tested.
                                        were collected for hormone assays                        females), increase                       30 gr).
                                        and western blot analysis,                               expression of StAR and
                                        respectively.                                            cyp11A1 (female).
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<pre> Annex                                                                                                                                                             Bisphenol A | page 45 of 51
 Reference              Analysed        Study design                          Tested doses        Critical effect(s) observed Studied          Lowest effect level     Comments/Main limitations
                        species and sex                                                                                          generation                            for risk assessment
 Menard et al. (2014)80 Rat, female     Pregnant and lactating rats were      0, 0.5, 5, 50 µg/kg Increase of anti-OVA IgG       F1            0.5 µg/kg bw/d          Oral allergy/tolerance model
                                        treated by gavage from GD15 to        bw/d                titers at all BPA dosages in                                         limited relevance for workers;
                                        weaning (D21). At D45, Immune                             OVA-tolerized rats, and at 5                 (for IgG titers)        only for one parameter all
                                        parameters of offspring were tested                       µg/kg/d in OVA-immunized                                             doses were tested; limited
                                        after oral tolerance or immunisation                      rats compared to control                                             dose response information.
                                        to OVA.                                                   rats treated with vehicle. In
                                                                                                  BPA-treated and OVA
                                                                                                  tolerized rats, increased
                                                                                                  anti-OVA IgG titers were
                                                                                                  associated with higher IFN
                                                                                                  secretion by the spleen,
                                                                                                  and an increase of
                                                                                                  activated T-cells. Also,
                                                                                                  when BPA-treated
                                                                                                  OVA-tolerized rats were
                                                                                                  orally challenged with OVA,
                                                                                                  colonic inflammation
                                                                                                  occurred, with neutrophil
                                                                                                  infiltration, increased IFN
                                                                                                  and decreased TGF.
 Menard et al. (2014)81 Rat, female     Rats were fed with BPA for GD15 to 0 and 5 µg/kg bw/d     Decrease in OVA-induced        F1            5 µg/kg bw/d            Oral allergy/tolerance model
                                        weaning. Immune parameters of                             IFN secretion and T-cells/                                           limited relevant for workers;
                                        offspring were tested after oral                          dendritic cells in spleen and                                        only one dose tested.
                                        tolerance or immunisation to OVA;                         mesenteric lymph nodes;
                                        or infection by intestinal nematodes.                     1.5-fold increase in N.
                                                                                                  brasiliensis living larvae in
                                                                                                  the intestine of
                                                                                                  BPA-exposed rats
                                                                                                  compared to controls.
 Moore-Ambriz           Mouse, young    Mice were exposed to 50 µg/kg         0 and 50 µg/kg bw   No effect on parameters        No study on   50 µg/kg bw/d           No effects reported that were
 (2015)41               adult, female   bw/d by gavage for a period of 3                          related to ovulation, but it   developmental                         considered adverse; only one
                                        reproductive cycles.                                      reduced the fraction of        effects.                              dose tested; limited number of
                                                                                                  fertilized oocytes after in                                          animals.
                                                                                                  vitro fertilisation or mating.
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<pre> Annex                                                                                                                                                             Bisphenol A | page 46 of 51
 Reference           Analysed        Study design                            Tested doses           Critical effect(s) observed Studied      Lowest effect level       Comments/Main limitations
                     species and sex                                                                                              generation                           for risk assessment
 NTP (2018)(draft)72 Rat, male and   CLARITY regulatory perinatal and        0, 2.5, 25, 250,       Few significant effects, not  F1         No clear exposure-        Study with extended dose
                     female          chronic study. BPA was                  2,500 and 25,000       clear whether these were                 related effects reported. range and various parameters,
                                     administered by oral gavage from        µg/kg bw/d             treatment related.                                                 conducted following FDA GLP
                                     GD 6 through the start of labor and                                                                                               regulations.
                                     then directly to pups from PND 1,       Positive control:      In the stop-dose group, at 2
                                     until 1 year or 2 years. In addition, a (0.05 and 0.5 µg/kg    years there was a statist
                                     stop-dose study arm was included        bw/d)                  significant increase in
                                     with animals dosed only until PND                              incidence of female
                                     21.                                                            mammary gland
                                     Data collected included body                                   adenocarcinoma (22% vs
                                     weights, litter parameters, age at                             6%) and adenoma and
                                     vaginal opening, vaginal cytology,                             adenocarcinoma combined
                                     clinical chemistry, sperm                                      (24% vs 8%) at 2.5 µg/kg
                                     parameters, organs weights and                                 bw/d.
                                     histopathology.
                                                                                                    In the continuous dose arm,
                                                                                                    there was an increase in
                                                                                                    female mammary gland
                                                                                                    atypical foci at 2.5 µg/kg
                                                                                                    bw/d (14% vs 0% (interim)
                                                                                                    and 15% vs 4%
                                                                                                    (termination)).
                                                                                                    There was a significant
                                                                                                    trend (p=0.037) for uterine
                                                                                                    stromal polyps in the interim
                                                                                                    sacrifice group of the
                                                                                                    continuous dose arm.
 Nygaard et al.      Mouse, males    Dams were exposed to BPA in             0, 10 or 100 mg/mL     At 100 mg/mL: increased       F1         20 mg/kg bw               Relevance airway allergy and
 (2015)82            and females     drinking water from time of mating      (equivalent to 0, 2 or eosinophil numbers in                                              model unclear; food tolerance
                                     and until the end of the lactation      20 mg/kg)              bronchoalveolar lavage fluid                                       not relevant for workers.
                                     period. Offspring were administered                            (BALF) and a trend of
                                     10 mg OVA intraperitoneally without                            increased OVA-specific IgE
                                     adjuvant at PND4 and 18. At                                    levels.
                                     PND25 they were challenged OVA
                                     intranasally. Analysis at PND30.
45        Health Council of the Netherlands | No. 2019/04                                                                                               2                                           47
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<pre> Annex                                                                                                                                                         Bisphenol A | page 47 of 51
 Reference              Analysed        Study design                        Tested doses       Critical effect(s) observed Studied      Lowest effect level        Comments/Main limitations
                        species and sex                                                                                      generation                            for risk assessment
 Patel et al. (2017)57  Rat, female     Part of CLARITY.                    0, 2.5, 25, 250,   Reduction of ovarian follicle F1         -                          Effects only borderline
                                        Rats were dosed with BPA from       2,500 and 25,000   numbers and reduction of                                            significant; decrease follicle
                                        GD6 until up to 1 year. Ovarian     μg/kg              sex steroid levels at some                                          count only observed at
                                        morphology and serum estradiol                         doses and time points.                                              PND21; no dose-response
                                        and progesteronea were analysed                                                                                            relationship.
                                        at PND1, 21, 90, and at 6 months
                                        and 1 year.
 Quan et al. (2016)42   Rat, male       Rats were exposed by gavage from 0, 1, 10 and 100 mg/  Effects on hormones,          F1         1 mg/kg bw/d               Limited or lack of dose-
                                        GD14-21 and analysed at PND21       kg bw              oxidative stress, inhibition                                        response; relevance of
                                        and levels of sex hormones and                         spermatogenesis,                         Sperm density and          parameters; relatively high
                                        reactive oxygen species,                               apoptosis.                               abnormalities              doses applied.
                                        expressions of proteins and genes
                                        in the Akt/mTOR, and mitochondrial
                                        apoptosis pathways were
                                        measured. Sperm quality was
                                        assessed at PND50.
 Rahman et al.          Mouse, analysed Mice were gavaged BPA from          0, 50 μg/kg, bw, 5 Inhibition sperm count,       F1         50 µg/kg bw/d              Adversity of most critical
 (2016)44               at PND 120,     GD7-14. At PND120, function,        and 50 mg/kg bw    motility parameters and                                             parameter unclear; no dose
                        male            fertility, related processes and                       intracellular ATP, reduced               Only for intracellular ATP response for intracellular ATP
                                        protein profiles of F1 spermatozoa                     litter size                              reduction and SOD2;        reduction; functional effects
                                        were assessed.                                                                                  other effects only at      only observed at high doses.
                                                                                                                                        higher doses (reduced
                                                                                                                                        litter size only observed
                                                                                                                                        at highest tested dose.
 Rebuli et al. (2015)73 Rat, males and  Part of CLARITY.                    0, 2.5, 25, and    No consistent effects of      F1         No significant/treatment   National Research Council
                        females         Animals were dosed by oral gavage 25,000 µg/kg bw/d    BPA were observed for any                related effects were       guidelines followed; limited
                                        from GD 6 and continued until                          endpoint, in either sex, at              observed.                  study power was noted.
                                        parturition, pups after birth until                    either age compared to
                                        postnatal day (PND) 21.                                vehicle controls.
                                        Behavioural assessments were
                                        performed either on PND25-27 or
                                        PND97-125, and included open
                                        field, elevated plus maze, and zero
                                        maze.
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<pre> Annex                                                                                                                                                             Bisphenol A | page 48 of 51
 Reference              Analysed        Study design                          Tested doses         Critical effect(s) observed Studied        Lowest effect level      Comments/Main limitations
                        species and sex                                                                                         generation                             for risk assessment
 Santamaria et al.      Rat, adult,     Low dose exposure to pregnant         0, 0.5 and 50 µg/kg  Abnormalities in ovaries     F1            0.5 µg/kg bw/d           Not clear whether effects
 (2016)45               female          rats via drinking water from GD9 to bw                     and follicles.                                                      should be considered adverse;
                                        weaning on PND21. Oestrous cycle                                                                      (calculated by authors   only one dose tested; effects
                                        was assessed in offspring treated                                                                     based on mean body       difficult to interpret.
                                        with BPA from PND45-90. At                                                                            weight dams).
                                        PND90, blood and ovaries from F1
                                        dams were obtained.
 Suglia et al. (2016)46 Mouse, male,    Pregnant mice were exposed            0 and 10 µg/mL in    Reduced body weight and      F1            1.8 mg/kg bw/d           Not clear whether effects
                                        during a foetal-perinatal period (10d drinking water       food intake; decreased                                              should be considered adverse;
                                        post coitum to PND31), body                                epididymal fat mass; effects               (specified by authors)   only one dose tested.
                                        weight, food intake, fat mass, and                         on gene expression in
                                        hypothalamic signals related to                            hypothalamus.
                                        anorexigenic control of food intake
                                        were analysed at day 78.
 Tarapore et al. (2016; Rat, male       Female rats were fed a diet with      0, 2.5, 25, 250 and  Impaired spermatogenesis,    F1            25 µg/kg bw/d            Only one dose tested with
 2017)47,48                             BPA before and during pregnancy.      2,500 µg/kg bw/d     measured as an increased                                            standard diet; limited number
                                        Different stages of spermatogenesis                        fraction of seminiferous                                            of animals; no pattern in the
                                        was analysed in, offspring at                              tubules with impaired                                               results.
                                        PND210.                                                    spermatogenesis at the
                                                                                                   round spermatid step 1.
 Tiwari and Vanage      Rat, male       Rats received BPA by gavage for       0, 0.01 and 5 mg/kg  Increased lipid peroxidation No study on   0.01 mg/kg bw/d          Not clear whether effects
 (2017)58                               6d, where after lipid peroxidation    bw/d                 and decreased activity       developmental                          should be considered adverse.
                                        and various antioxidant enzymes                            various antioxidants in      effects.
                                        were measured in bone marrow                               various cells/tissues.
                                        cells and blood lymphocytes, and in
                                        testis and epididymis.
 Van Esterik (2014;     Mouse, male and Mice were exposed during              8 doses (0-3,000 µg/ Reduction in tissue and      F1            Lowest BMDL05: 233       Not clear whether effects
 2015)49,50             female          gestation and lactation via pellet;   kg bw                body weights; physical                     µg/kg bw/d               should be considered adverse
                                        Offspring were followed for 20                             activity; biochemical                      (interscapular weight in (noted as non-toxic doses by
                                        weeks. Glucose tolerance test,                             parameters.                                females).                the authors).
                                        spontaneous locomotor activity,
                                        histopathology, clinical chemistry,                                                                   (dose was calculated by
                                        gene expression analysis and DNA                                                                      authors based on food
                                        methylation was performed.                                                                            consumption).
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<pre> Annex                                                                                                                                                                        Bisphenol A | page 49 of 51
  Reference                Analysed           Study design                         Tested doses          Critical effect(s) observed Studied             Lowest effect level        Comments/Main limitations
                           species and sex                                                                                              generation                                  for risk assessment
  Whitehead et al.         Mouse, foetus,     Pregnant mice fed 25 mg/kg diet      0 and 25 mg/kg diet   Increased number of            F1               4.8 mg/kg bw/d             Not clear whether effects
  (2016)52                 sex not reported   from GD7.5-18.5. Thereafter foetal                         islet-cell clusters, increased                  (recalculated using        should be considered adverse;
                                              pancreata were collected and                               glucagon expression in                          TGD, assuming bw of 30 only one dose tested.
                                              analysed for morphological                                 islets and numbers of                           gr).
                                              changes.                                                   glucagon-expressing
                                                                                                         islet-cell clusters.
  Xie et al. (2016)53      Mouse, male        Newborn male mice were               0 and 0.01, 0.1 and   Testis: signs of meiotic       No study on      100 µg kg bw/d             Model not relevant for workers.
                                              subcutaneously injected with BPA     5 mg/kg body weight   arrest (spermatogonia and      developmental                               Exposure route not relevant for
                                              on PND1-21. At PND22, histological                         spermatocytes with             effects.                                    workers; limited dose-
                                              analysis was performed on the                              markedly less round                                                        response.
                                              testes.                                                    spermatids), increased
                                                                                                         apoptosis, abnormal
                                                                                                         proliferation.
 Abbreviations: AST - aspartate aminotransferase; ALT - alanine aminotransferase; BPA - bisphenol A; FDA - Food and Drug Administration; GD - gestation day; GLP - Good Laboratory Practice; GLUT 1,3 -
 transporter 1, 3; GPx - glutathione peroxidase; MDA - malondialdehyde; NO - nitric oxide; PND - post natal day; OVA - ovalbumine; StAR - steroidogenic acute regulatory protein; SOD2 - superoxide dismutase.
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<pre>                                                                                                                                                                         Bisphenol A | page 50 of 51
 The Committee                                                                                        Observers:
 • F.G.M. Russel, Professor of Pharmacology and Toxicology, Radboud University Medical Center,        •  H. Stigter,  Occupational Physician, Expertise Centre, Ministry of Social Affairs and Employment
   Nijmegen, chairman                                                                                 •  D. Theodori, Social and Economic Council, The Hague
 • P.J. Boogaard, Professor of Environmental Health and Human Biomonitoring, Wageningen
   University and Research Centre, and Toxicologist, Shell International BV, The Hague                Scientific secretary:
 • R. Houba, Occupational Hygienist, Netherlands Expertise Centre for Occupational Respiratory        •  S.R. Vink, Health Council of the Netherlands, The Hague
   Disorders (NECORD), Utrecht
 • E.D. Kroese, Toxicologist, TNO Innovation for Life, Zeist
 • F.C. Kuper, Toxicologic Pathologist, Utrecht
 • H. van Loveren, Professor of Immunotoxicology, Maastricht University, Maastricht
 • I.M.C.M. Rietjens, Professor of Toxicology, Wageningen University and Research Centre,
   Wageningen
 • G.B.G.J. van Rooy, Occupational Physician/toxicologist, Arbo Unie Expert Centre for Chemical Risk
   Management, and Radboud UMC Outpatient Clinic for Occupational Clinical Toxicology, Nijmegen
 • L.A. Smit, Epidemiologist, Institute for Risk Assessment Sciences, Utrecht
 • R.C.H. Vermeulen, Professor of Environmental Epidemiology and Exposome Science, Institute for
   Risk Assessment Sciences, Utrecht
 • A.H. Piersma, Professor of Reproductive and Developmental Toxicology, Utrecht University, and
   National Institute for Public Health and the Environment, Bilthoven, structurally consulted expert
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<pre> The Health Council of the Netherlands, established in 1902, is an independent scientific advisory body. Its remit is “to advise the government and
 Parliament on the current level of knowledge with respect to public health issues and health (services) research...” (Section 22, Health Act).
 The Health Council receives most requests for advice from the Ministers of Health, Welfare and Sport, Infrastructure and Water Management, Social
 Affairs and Employment, and Agriculture, Nature and Food Quality. The Council can publish advisory reports on its own initiative. It usually does this in
 order to ask attention for developments or trends that are thought to be relevant to government policy.
 Most Health Council reports are prepared by multidisciplinary committees of Dutch or, sometimes, foreign experts, appointed in a personal capacity.
 The reports are available to the public.
 This publiation can be downloaded from www.healthcouncil.nl.
 Preferred citation:
 Health Council of the Netherlands. Bisphenol A. Health-based recommendation on
 occupational exposure limits.
 The Hague: Health Council of the Netherlands, 2019; publication no. 2019/04.
 All rights reserved
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