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MASTER THESIS zur Erlangung des akademischen Grades „Master of Science in Engineering“ im Studiengang „Technisches Umweltmanagement und Ökotoxikologie“

Bisphenol A: Environmental Fate and Behaviour, Human Health Effects – Implications for EU-chemical Policy Ausgeführt von: Mag.a Christina Hartmann Personenkennzeichen: 1110332006

1. Begutachter: Dr. Mag. Thomas Jakl 2. Begutachterin: Dr. Maria Uhl, MTox

Wien, 04.12.2012

Eidesstattliche Erklärung „Ich erkläre hiermit an Eides statt, dass ich die vorliegende Arbeit selbständig angefertigt habe. Die aus fremden Quellen direkt oder indirekt übernommenen Gedanken sind als solche kenntlich gemacht. Die Arbeit wurde bisher weder in gleicher noch in ähnlicher Form einer anderen Prüfungsbehörde vorgelegt und auch noch nicht veröffentlicht. Ich versichere, dass die abgegebene Version jener im Uploadtool entspricht.“

Ort, Datum

Unterschrift

Kurzfassung Bisphenol A (BPA) ist eine Umweltchemikalie, die in einer Vielzahl von Konsumprodukten ihren Einsatz findet. Weltweit wird BPA in sehr großen Mengen produziert und unter anderem für die Produktion von Polykarbonaten (PC), Polyvinylchlorid (PVC) und Epoxidharzen verwendet. Durch seine chemische Struktur ist es möglich, dass BPA aus den Produkten migrieren kann, und somit in Nahrungsmittel, Getränke, Luft und Wasser übergehen kann. Dadurch erfolgt eine Belastung des Menschen über diverse Aufnahmewege. BPA zählt zu den endokrin wirksamen Chemikalien und kann unterschiedliche Effekte auslösen. In verschiedenen Verordnungen und Richtlinien wird BPA gesetzlich reguliert, ist nach der CLP-Verordnung klassifiziert, steht im Verdacht, die Fertilität zu schädigen und ist gesundheitsschädlich. In den letzten Jahrzehnten wurden eine Vielzahl von wissenschaftlichen Studien durchgeführt, um die Effekte von BPA auf Tiere und Menschen zu untersuchen, sein Umweltverhalten, seine Toxizität auf Menschen sowie das Ökosystem. Das tatsächliche Risiko, welches durch die aktuelle Belastung mit BPA besteht, ist umstritten. In Diskussion stehen beispielsweise die Dosis-Wirkungsbeziehung, Effekte im Niedrigdosis-Bereich und deren Prozesse, Belastungspfade, das Ausmaß der Exposition, die Inaktivierung von BPA im Organismus, die Übertragbarkeit von Ergebnissen aus Studien an Tieren auf den Menschen und deren Relevanz für die Risikobewertung, oder die Qualität und Aussagekraft diverser wissenschaftlicher und toxikologischer Studien. Ziel dieser Arbeit ist es, einen detaillierten Überblick über den REACH-Prozess, die gesetzlichen Bestimmungen zu BPA, die existierenden Meinungsstreitigkeiten und wissenschaftlichen Stellungnahmen, sowie das Umweltverhalten und die Toxizität von BPA zu geben.

Schlagwörter: Bisphenol A, Chemikalienpolitik, REACH, Gesundheit, Umwelt

3

Abstract Bisphenol A (BPA) is an industrial chemical and widespread used in many consumer products. It is manufactured in very high quantities and is used amongst others for the production of polycarbonate (PC), polyvinyl chloride (PVC) and epoxy resins, and is able to leach into food, beverages and air. Thus, humans are exposed to BPA through different routes of exposure. BPA is identified as an endocrine disrupting chemical and is able to lead to various adverse health effects. It is regulated in various directives and regulations, and classified and labelled according to the CLP regulation as a substance that is suspected to damage fertility and be health hazard. In the recent decades, a multitude of efforts have been performed to assess the health effects of BPA in animal studies and studies in humans, the fates and pathways of BPA in the environment, and the its toxicity to humans, wildlife and laboratory animals, as well as its ecotoxicology. Although (or maybe because of) the availability of thousands of studies on BPA, in science, as well as in governments and agencies, many controversies about BPA exist about the dose-response curves, low-dose effects and action, routes of exposure, the inactivation of BPA in the body, the relevance of animal studies for human risk assessments, or the utility, quality and reliability of scientific and toxicological studies. Aim of this thesis is to give a detailed overview on the REACH process, the legislation of BPA and its chemical policy, the existing controversies, scientific opinions, the environmental fates and pathways, as well as the toxicity of BPA to humans, wildlife and ecotoxicology.

Keywords: Bisphenol A, Chemical Policy, REACH, Human Health, Environment

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Danksagung

Mein herzlichster Dank gebührt

Herrn Dr. Mag. Thomas Jakl als Erstbetreuer meiner Masterarbeit für seine überaus freundliche und fachkundige Unterstützung und Begleitung, Frau Dr. Maria Uhl, MTOX als Zweitbetreuerin und Projektleiterin der UM-MuKi-Studie für die Unterstützung, Begleitung und die Bereitstellung der Daten, Frau Ing. Andrea Sitka für die Probenanalysen im Zuge der UM-MuKi-Studie, Herrn Ing. Wolfgang Raffesberg für die Probenanalysen sowie die regelmäßigen fachlichen Diskussionen, sowie Herrn Dr. Stefan Weiß als Arbeitsgruppenleiter für seine Unterstützung.

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Table of Contents 1

INTRODUCTION ........................................................................................................ 8

1.1

Problem definition .................................................................................................................... 8

1.2

Objectives ................................................................................................................................ 9

2

GENERAL INFORMATION ON BPA ..........................................................................10

2.1

Physical and chemical properties .......................................................................................... 11

2.2

Biochemical properties........................................................................................................... 11

2.3

Production, usage and alternatives ....................................................................................... 12

3

ENVIRONMENT ........................................................................................................16

3.1

Environmental Releases ........................................................................................................ 16

3.2

Environmental Fates and Pathways ...................................................................................... 18

3.3

Ecotoxicology ......................................................................................................................... 25

4

HUMAN HEALTH .......................................................................................................35

4.1

Human exposure .................................................................................................................... 35

4.2

Metabolism and Toxicokinetics .............................................................................................. 38

4.3

Toxicity ................................................................................................................................... 38

5

CHEMICAL POLICY ..................................................................................................47

5.1

Former Chemicals Legislation in the EU ............................................................................... 47

5.2

REACH Regulation ................................................................................................................ 48

5.3

Endocrine disruptors legislation in the EU ............................................................................. 59

5.4

Legislation of BPA in the European Union ............................................................................ 66

5.5

BPA under REACH ................................................................................................................ 71

5.6

International Legislation of BPA ............................................................................................. 73

5.7

Opinions by Stakeholders and Authorities ............................................................................. 75

5.8

Controversies on BPA............................................................................................................ 81 6

6

UM-MUKI STUDY ......................................................................................................86

6.1

Human biomonitoring ............................................................................................................. 86

6.2

Methods and Materials........................................................................................................... 88

6.3

Results and discussion .......................................................................................................... 88

7

DISCUSSION.............................................................................................................91

8

METHODS .................................................................................................................93

9

REFERENCES ..........................................................................................................94

ANNEX A: SUMMARY OF THE ANNEXES OF REGULATION (EC) NO. 1907/2006 (REACH) .........................................................................................................................121

7

1 Introduction 1.1 Problem definition Bisphenol A (BPA; 2.2-bis(4-hydroxyphenyl)propane) is an industrially used chemical and is produced worldwide at high annual quantities. BPA is used as a monomer in polycarbonate and epoxy resin production, as well as in polyvinyl chloride manufacture and processing, and thermal paper production. [EC, 2010] Thus, medical equipment, dental inlays, plastic bottles, digital media, household appliances, microwave dishes, and other products that are made of polycarbonate contain BPA.

Epoxy resins are used in

adhesives, can and powder coatings, floorings or industrial protective coating, and a multitude of other products. [German Federal Environment Agency, 2010a; IHCP, n.r.] Because of its chemical structure, BPA is able to migrate from plastic food contact materials, and other consumer products into food, beverages or air, and is also found in house dust. Thereby, humans are exposed to BPA. It can be detected in breast milk, urine, blood, and other body fluids and tissues. [Vandenberg et al., 2007] Many studies have investigated the effects on environment and human health as well as the toxicity of BPA and got conflicting results. BPA is an endocrine disrupting chemical and has weak estrogenic effects. It has been shown that BPA causes a variety of effects on development and reproduction in animals, even at low doses. [Dekant and Völkel, 2008] It is able to stimulate various molecular endpoints and is associated with several diseases. [Newbold et al., 2009] Potential risks arising from human exposure levels are scientifically discussed in a very controversial way. [Dekant and Völkel, 2008]

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1.2 Objectives Aim of this thesis is to give a detailed overview on the effects and toxicity of BPA to environment and human health. It is supported by an outline of its legislation, classification under REACH and relating problems, and of the scientific controversies. Research topics Following research questions are examined: -

What is Bisphenol A and how is it produced and used?

-

What is the legislation related to BPA?

-

How does REACH work?

-

What is the classification of BPA according to GLP?

-

BPA under REACH

-

What is the state of the discussion about BPA and its health effects?

-

What are the environmental fates and pathways?

-

How severe is the environmental and human toxicity of BPA?

9

2 General Information on BPA IUPAC Name: 2,2-bis(4-hydroxyphenyl)propane EC Name: 4,4´-isopropylidenediphenol

Figure 1. Chemical structure of Bisphenol A

Table 1. General Information of Bisphenol A [EC, 2010; ECHA, 2012a] CAS No.

EC No.

Annex I

Annex I Status

Index No.

ATP inserted

European

ATP last update

Priority List No.

80-05-7

201-245-8

604-030-00-0

67/548/EEC

19

annex I

30

3

Synonyms: BPA (abbreviation); 2,2-Bis(4-hydroxyphenyl)propane; 2,2-Bis(p-hydroxyphenyl)propane; p,p’-Isopropylidene-bisphenol; p,p’-Isopropylidene-di-phenol; Phenol, 4,4’Isopropylidene-di; Diphenylol Propane; Parabis (Trademark); Bis (4-hydroxyphenyl) dimethyl methane; Bis (4-hydroxyphenyl)propane; Dian (Trademark); Dimethylmethylenep,p’-di-phenol;

Dimethyl

Bis(p-hydroxyphenyl)methane;

4,4’-Dihydroxy-2,2’-diphenyl

propane; 4,4’-Dihydroxydiphenyldimethyl methane; 4,4’-Dihydroxydiphenyl propane; ß-Dip-Hydroxyphenyl diphenyl propane;

propane;

propane;

p,p’-Dihydroxydiphenyldimethyl

2,2’-(4,4’-Dihdroxydiphenyl)

2,2’-Di(4-hydroxyphenyl)

propane;

propane;

methane;

p,p’-Dihydroxy-

4,4’-Dihydroxydiphenyl-2,2’-

2,2’-Di(4-phenylol)

propane;

4,4’-

Isopropylidene bisphenol; 4,4’-(1-methylethylidene)bisphenol [ECHA, 2012a]

Trade Names: Bisphenol A; BPA; Diphenylol propane; DPP; Bisphenol A ER grade Flake; PARABIS(R) Resin intermediate; Bisphenol A - Polycarbonate grade [EC, 2010; ECHA, 2012a]

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2.1 Physical and chemical properties Table 2. Substance type, chemical and physical information of Bisphenol A [United Kingdom, 2008; EC, 2010; ECHA, 2012a] Substance Type

Organic; mono constituent substance

Chemical Information

Molecular Formula: C15H16O2 Molecular Weight: 228.29 g/mol

Physical Information

State: white solid flakes or powder at normal temperature (20°C) and pressure 1013 hPa) Density: 1.2 kg/m3 (at 25°C) Water solubility: 300 mg/L logKow: 3.4 Flash point: 207 °C Melting point: ≥ 156°C Freezing Point: ≥ 157°C Boiling Point: 360°C (at atmospheric pressure=1013 hPa) Flammability: 532°C Self-ignition temperature: 510°C

At environmentally relevant conditions (20°C, 1013 hPa) BPA is an organic white solid odourless substance [Ashford, 1994]. BPA has its melting point at ≥ 156°C, its freezing point at ≥ 157°C [Sax and Lewis, 1996] and the boiling point is 360°C at 1013 hPa [ECHA, 2012b]. At 25°C the relative density is 1.2 g/cm3 [Sax and Lewis, 1996] and the vapour pressure is 4.12E-9 hPa [ECHA, 2012c]. The water solubility of BPA at 25°C is ca. 300 mg/L [ECHA, 2012d]. The flash point (1013hPa) is 227°C and the auto flammability (autoignition temperature) is 510°C at 1013 hPa [Gerhartz, 1991]. BPA is a stable substance and is not corrosive to metals [ECHA, 2012e].

2.2 Biochemical properties BPA was synthesised for the first time in 1891 by A.P. Dianin. In the 1930s it was investigated and tested for its estrogenic properties because of the interest in synthetic estrogens. [Vandenberg et al., 2009] In 1936, the estrogenic properties were reported by 11

Dodds and Lawson for the first time. [Rykowska and Wasiak, 2006] BPA is synthesised by combination of two phenol equivalents and one acetone equivalent in presence of an acidic ion-exchange resin as catalyst and contains therefore two phenol functional groups. The chemical structure of BPA has two benzene rings and two 4,4’-OH-substitues (Figure 1). BPA is able to bind to the estrogen receptors (ER) ERα and ERβ, with a 10-fold lower affinity to ERα. [Vandenberg et al., 2009] For detailed information of the estrogenic activity of BPA see Chapter 4.3.4.

2.3 Production, usage and alternatives BPA is manufactured usually in high purity by an alkaline or acidic catalysed condensation of phenol and acetone. It is used for polycarbonate (PC) production (71.1%), epoxy resin production (25.0%), phenoplast resins, unsaturated polyester resin production, can coating manufacture, polyvinyl chloride (PVC) production and processing, thermal paper production,

polyols/polyurethane

production,

modified

polyamide

production,

tyre

manufacture, brake fluids and others. [EC, 2010] Depending on the manufacturer, BPA has a purity of 99.0 – 99.8%. The impurities include phenol, ortho- and para-isomers of BPA and water. [EC, 2008a] Figure 2 shows the synthesis process of PC resins.

Figure 2. Synthesis process of polycarbonate resins [Rykowska and Wasiak, 2006]

The advantages of PC plastics are a light weight, a high tensile strength, durability, high elasticity, a high melting point as well as a high vitrification temperature, which is one of the reasons of the wide use. [Rykowska and Wasiak, 2006] PC products have to be marked

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with a recycling code1 (Figure 3). This recycling code is usually used for PC, but also new bio-based plastics may be labeled with this code [Erler and Novak, 2010].

Figure 3. Recycling-Code of Polycarbonate [Erler and Novak, 2010]

Examples for polycarbonate products that can contain BPA are medical equipment, water bottles, electrical equipment, eyeglass lenses, consumer electronics, casings of mobile phones or computers, digital media (e.g. CDs, DVDs, Blu-ray DiscsTM), household appliances, sports safety equipment, automobiles and microwave dishes. Epoxy resins are used in floorings, adhesives, can coatings, powder coatings or industrial protective coatings for example. [German Federal Environment Agency, 2010a; IHCP, n.r.]

Table 3. BPA use pattern data in the EU for the years 1996-1999 (cf. [EC, 2008a])

BPA use pattern data

Tonnes per year (percentage of the EU consumption)

Total EU consumption

685,000 (100%)

PC production

486,880 (71.1%)

Epoxy resin production

171,095 (25.0%)

Phenoplast cast resin processing

8,800 (1.3%)

Unsaturated polyester resin production

3,000 (0.4%)

Can coating production

2,460 (0.4%)

Use PVC production and processing

2,250 (0.3%)

Alkyloxylated BPA manufacture

2,020 (0.3%)

Thermal paper production

1,400 (0.2%)

Polyols/Polyurethane manufacture

950 (0.1%)

Modified polyamide production

150 (0.1%)

Tyre manufacture

110 ( 290 nm directly resulting in photochemical alteration. The extent of the photolysis depends on the conditions of the water like pH, water turbulence and turbidity, among others and the amount of the sunlight. The process of the photooxidation is the degradation of substances through oxidants (mainly hydroxyl radicals). Both processes occur in water and in atmosphere. [Staples et al., 1998] Wang et al., 2006 investigated the photodegradation of BPA in simulated lake water that contained algae, humic acid and ferric ions. The BPA photodegradation was enhanced by algae, humic acid and ferric ions, whereas the photodegradation in water was greater with the combination of raw algae and humic acid and/or ferric ions than water containing raw algae. [Peng et al., 2006] In an experimental study of photodegradation of BPA in water, Nakatani et al., 2003 had shown that in absence of nitrate in the water, BPA became only less degraded. This suggests that the direct photoreaction is not significant under sunlight exposure. Compared with this direct photoreaction, BPA was degraded in a faster way in the presence of nitrate. The authors concluded that the degradation of BPA is caused by nitrate photolysis.

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Additionally these findings suggest that BPA gets transformed into other organic compounds. [Nakatani et al., 2003] In air, the photooxidation half-life of BPA basing on hydroxyl radical reactions was calculated at a range of 0.74 to 7.4 hours. This calculation was made with the so-called Atmospheric Oxidation Program which is basing on structure-properties of BPA. [Cousins et al., 2002]

3.2.3 Bioaccumulation Bioaccumulation is the accumulation of (a) substance(s) in organism tissues through any route of exposure. It is the combination of bioconcentration and biomagnification: [USGS, 2011b; Utah State University, n.r.] Bioconcentration is defined as a process that leads to a higher concentration of a substance in an exposed organism in comparison to the concentration in an environmental media. Thus, it is the process of accumulation of substances through non-dietary routes in aquatic organisms, which means an uptake of a substance from water via respiratory surface and/or via skin. The ratio of the concentration of a substance in organisms to the concentration in its surrounding medium is measured as the so-called bioconcentration factor (BCF). [Utah State University, n.r.; USGS, 2011c] Biomagnification is defined as the uptake via food. [Utah State University, n.r.] The experimental conditions of the estimation of BCF in aquatic species are reported in the OECD 305 guideline. According to REACH, the estimated BCFs of a substance are used for the classification and labelling of a substance, for the PBT and vPvB assessment and the chemical safety assessment. The thresholds for the PBT or vPvB identification of a substance are for bioaccumulative (B) a BCF > 2000 L/kg (or 3.3 in log unit) and for very bioaccumulative (vB) a BCF > 5000 L/kg (3.7 in log unit). [Istituto di Ricerche Farmacologiche Mario Negri, n.r.] In several studies the bioaccumulation of BPA was investigated: Li et al., 2009 investigated in their study the bioaccumulation and removal capability of the marine microalga Stephanodiscus hantzschii to BPA. They reported an EC50 of BPA after 95 h of 8.65±0.26 mg/L. In this microalga, the cell number and chlorophyll a content significantly decreased with increasing BPA levels at concentrations higher than 3 mg/L. BPA was bioaccumulated and biodegraded by cells. Those results suggested that Stephanodiscus hantzschii can be used for BPA remove from contaminated waters. [Li et al., 2009] 21

Lindholst et al., 2001 investigated the uptake, metabolism and excretion of BPA in juvenile rainbow trout (Oncorhynchus mykiss). The estimated BCFs for this fish ranged between 3.5 and 5.5 for plasma, liver and muscle at a concentration of 100 µg/L water. [Lindholst et al., 2001] Staples et al., 1998 reported BCFs between < 20 and 196 L/kg for BPA. [Staples et al., 1998] Lee

et

al.,

2004

investigated

the

BPA

and

nonylphenol

concentrations

and

bioaccumulation in a marine fish after exposure of BPA and nonylphenol in seawater. The estimated BCFs of BPA were 38.14 ± 21.28 L/kg (mean ± SD) after 2 days of exposure and at an exposure concentration of 70 µg/L, and 13.34 ± 5.14 L/kg after 1 day of exposure at a concentration of 700 µg/L. [Lee et al., 2004] Liu et al., 2011 studied the distribution and bioaccumulation of various steroidal and phenolic EDCs in different tissues of wild fish species in a lake in China. BPA was found in muscle tissue with a maximal concentration of 83.5 ng/g dry weight. The highest concentrations were found in the liver. The BCFs ranged between 29 and 49. [Liu et al., 2011] Studies show that the BPA has a low potential of bioaccumulation in aquatic organisms. Thus, it is not classified as a bioaccumulative substance. [American Chemistry Council, 2003-2012a]

Octanol-Water Partition Coefficient The octanol-water partition coefficient (KOW) is defined as the ratio of the concentration of an organic substance in a known volume of n-octanol to its concentration in a known volume of water after reaching the equilibrium. The KOW is used as an indicator of an organic substance’s tendency to adsorb to soil. [USGS, 2011d] By the KOW, the bioconcentration potential of a substance can also be measured. At a log KOW a substance have a medium low affinity for an environmental compartment. Under a log KOW under 3, the substance is not bioaccumulative. [The Edinburgh Centre for Toxicity, n.r.] For BPA, the log KOW is 2.2 – 3.4. [Doerge and Fisher, 2010]

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3.2.4 Transport und distribution Atmospheric transport BPA is ubiquitous in the atmosphere. Fu and Kawamura, 2010 investigated BPA concentrations in atmospheric aerosols from different regions around the world. Measured concentrations ranged between 1 and 17400 pg/m3. In urban regions, they found positive associations between the BPA concentrations and 1,3,5-triphenylbenzene, which is an indicator for burned plastics. This indicates that the open burning of domestic waste containing plastics may be a significant source for BPA in the atmosphere. The highest BPA concentrations in aerosols were measured in metropolises in India. The investigation of marine aerosols showed that the highest BPA levels were estimated at coastal regions. Asia is a strong BPA emitter and by the westerly winds BPA can be transported long distances. In the Polar Regions BPA was also found, whereat in dark winter there were higher levels than in the early summer because the Arctic is acting as a cold sink. This suggests the atmospheric transport of BPA over long distances from Europe, Asia and North America to the Arctic. [Fu and Kawamura, 2010]

Water transport The environmental releases of BPA are mainly to surface waters. Cousins et al., 2002 performed a long-range transport model. Based on this model, BPA may be transported some 100 kilometres downstream from the point source. This estimated distance bases on the distance the water in a river will flow in the time in which BPA takes to be degraded to 50% of its initial concentration. In comparison, the transport in air is < 100 kilometre and thus much lower, because of the low vapour pressure of BPA. [Cousins et al., 2002]

Adsorption on soil and sediments Zeng et al., 2006 performed adsorption experiments on BPA to estimate the adsorption behaviour on sediments. They collected sediment samples from the Xiangjang River in China. They found out that the BPA adsorption process is an exothermic reaction. If the sediment concentration increases, the amount of adsorbed BPA on sediment decreases and increases with an increasing ironic concentration. In acidic sediments the amount of adsorbed BPA decreases with an increasing pH value. [Zeng et al., 2006]

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Ying and Kookana, 2005 estimated the soil-water distribution coefficients (Kd)2 on 4 different soils of various EDCs such as BPA, which had the lowest Kd values with a mean of 20. Additionally they found out, that the Kd values of all measured compounds increased with the soils organic carbon contents. They demonstrated that aerobic conditions fit the degradation more than anaerobic conditions. BPA adsorb to the organic matter in the soils and gets degraded rapidly under aerobic conditions. They reported that they are not persisting in aerated soils but in anaerobic soils. [Ying and Kookana, 2005] Höllrigl-Rosta et al., 2003 determined a soil organic carbon-water partition coefficient (KOC)3 of 890 L/kg. [Höllrigl-Rosta et al., 2003] Ying et al., 2003 investigated the sorption and degradation of EDCs including BPA in the laboratory by using groundwater and sediment from an aquifer from South Australia. The estimated sorption coefficient Kd of BPA was 3.89 L/kg and the KOC was determined to 778 L/kg. Under aerobic conditions BPA remained unchanged in the aquifer material. They calculated a retardation factor (Rf) for the BPA transport in groundwater of 8, which means that BPA is transported 1/8 of the velocity of the groundwater velocity. [Ying et al., 2003]

Persistence Cousins et al., 2002 provided models with inputs of environmental half-lives of BPA for air, water, soil and sediment. The half-time of a substance in a media is used for the assessment of the persistence of a chemical in the environment. The major loss routes of BPA are reaction in water (29.6%) and soil (56.8%). The predicted overall residence time of BPA in the model is 6.3 days, the reaction residence time 6.6 days and the advection residence time 136 days. Thus, BPA is not predicted to be persistent in the environment. [Cousins et al., 2002]

Henry’s Law constant The equilibrium of dilute substances of volatile, soluble substances between gas and liquid (water) is characterised by the Henry’s Law Constant. [U.S. EPA, 2012c] For BPA the calculated Henry´s law constant H is 3.12E-7 mPa*m3/mol at a temperature of 25°C and

2

The Kd describes the soil adsorption of a substance. The larger the K d the greater is the binding capacity of a substance. Kd > 5 has a high binding capacity in soil. [The Edinburgh Centre for Toxicity, n.r.] 3

The KOC describes the adsorption of a substance to organic carbon in soil. K OC > 500 has a high binding

capacity. [The Edinburgh Centre for Toxicity, n.r.] 24

atmospheric pressure (1013 hPa). Thus, the potential of BPA to volatilize from water or soil is very low. [ECHA, 2012i]

3.2.5 Summary of the environmental fates and pathways In air, the phototransformation of BPA occurs in a fast way [EC, 2003], and because of its chemical structure, the hydrolysis is not expected under environmental conditions [REACH consortium, 2011]. The biodegradation of BPA in water occurs rapidly [Dorn et al., 1987; Ying and Kookana, 2005; EC, 2010]. BPA is not persistent [Klečka et al., 2001; Cousins et al., 2002] or bioaccumulative in the environment [American Chemistry Council, 20032012a]. Additionally, BPA has no high binding capacity to organic carbon in soil. [HöllriglRosta et al., 2003] However, studies have shown that BPA is ubiquitous in atmosphere and becomes transported over long distances. Relationships between BPA concentrations and industrialized regions were shown in several studies. [Fu and Kawamura, 2010]

3.3 Ecotoxicology 3.3.1 Toxicity Aquatic toxicity Acute In a 14-day short-term toxicity test of BPA in zebrafish (Danio rerio) under semi-static conditions according to OECD guideline 204 the animals were exposed to BPA concentrations between 0.1 and 10 mg/L. Assessed endpoints were mortality and other visible effects on behavior and appearance. A 14-day LOEC of 10.15 mg/L and a 14-day NOEC of 3.2 mg/L were derived. [ECHA, 2012k] Mihaich et al., 2009 investigated the acute toxicity level of to BPA in aquatic invertebrates and compared their results with others in the scientific literature. Figure 6 shows the investigated NOEC values and EC50 values in various species. The EC50 values are ranging between 1.1 (Mysid, Mysidopsis bahia) and 10.2 mg/L (Water flea, Daphnia magna) and the NOEC between 0.51 and 4.1 mg/L, respectively. Additionally they investigated EC50 levels in aquatic plants and diatoms, which are ranging between 1.0 mg/L in Marine diatom (Skeletonema costatum) and 20 mg/L in Duckweed. (Lemna gibba). [Mihaich et al., 2009] 25

Figure 6. Invertebrate aquatic acute toxicity to BPA [Mihaich et al., 2009, p. 1397]

Various investigations derived by industry on the acute aquatic toxicity of BPA were present in the IUCLUD Dataset4 of the EC. In the following some of the reported toxicity values (only investigations under GLP conditions were used) are summarized [EC, 2000]: Marine fish (Menidia menidia):

9.4 mg/L (96-h LC50)

Fresh water fish (Pimephales promelas):

4.8 mg/L (96-h LC50)

Fresh water fish (Brachydanio rerio):

9.9 mg/L (96-h LC50)

Aquatic Invertebrates (Daphnia magna):

10.2 mg/L (48-h EC50)

Aquatic Invertebrates (Mysidopsis bahia):

1.1 mg/L (96-h LC50)

Algae (Selenastrum capricornutum):

2.7-3.1 mg/L (96-h EC50)

Algae (Skeletonema costatum):

1 mg/L (96-h EC50)

4

The IUCLID Dataset is a dossier basing on data by the European Chemicals Industry according to the “Council Regulation (EEC) No. 793/93 on the Evaluation and Control of the Risks of Existing Substances”. The available data in the IUCLID Dataset was not evaluated by the EC. [EC, 2000] 26

Chronic Mihaich et al., 2009 performed a 42-day chronic amphipod study exposing Amphipods (Hyalella azteca) to nominal BPA concentrations. They reported a LC50 of 0.78 mg/L, a LOEC of 1.1 mg/L and a NOEC of 0.49 mg/L. For chronic toxicity in invertebrates, various NOECs between 0.025 mg/L in Snail (Marisa cornuarietis) and > 3.16 mg/L in Water flea (Daphnia magna) were reported. LOECs ranged from 0.46 mg/L in Hydra (Hydra vulgaris) to 160 mg/L in Sponge (Heteromyenia sp.) (Figure 7). [Mihaich et al., 2009]

Figure 7. Invertebrate aquatic chronic toxicity to BPA [Mihaich et al., 2009, p. 1398]

Segner et al., 2003 investigated endocrine-disrupting effects in aquatic invertebrates and vertebrates. Zebrafish (Danio rerio) were exposed to BPA concentrations of 94 to 1,500 µg/L. The measured LOEC (endpoints: vitellogenin induction and gonad histology changes) was 375 µg/L and the NOEC 188 µg/L. For juvenile growth, time to spawning, fertilization success, eggs per female and mating behavior, the LOEC was 1500 µg/L and the NOEC 750 µg/L. [Segner et al., 2003; EC, 2010] Reported in the IUCLID Dataset, in aquatic invertebrates (Water flea, Daphnia magna) a 21-d NOEC, LOEC and EC50 value of > 3.2 were estimated. [EC, 2000]

Sediment toxicity Acute In a study of the acute toxicity of BPA to the amphipod Mud shrimp (Corophium volutator) in saltwater sediment published in 1999, the calculated 10-d LC50 values were 1.4 mg/L (46 mg/kg dw) for acetone spiked tests and 1.6 mg/L (60 mg/kg dw) for directly spiked tests. 27

The estimated 10-d EC50 values were 1.1 mg/L (31 mg/kg dw) for acetone spiked tests, and 1.3 mg/L (36 mg/kg dw) for directly spiked tests. [EC, 2010] Chronic In a 28-day chronic estuarine amphipod study Leptocheirus plumulosus was exposed to nominal BPA concentrations in natural marine sediment. The measured LOEC was 78 mg/kg and the NOEC was 32 mg/kg for amphipod survival, and > 32 mg/kg and 32 mg/kg for amphipod growth, respectively. The estimated LC50 for 28-days of survival was 63 mg/kg, and the EC50 was 60 mg/kg. [ECHA, 2012j]

Terrestrial toxicity Staples et al., 2010 performed a 28-d reproduction study in potworms (Enchytraeid sp.) and springtails (Collembolan sp.) in soil. For potworms, they calculated a 28-d NOEC of 100 mg/kg soil dw and a 28-d LC50 value of > 100 mg/kg soil dw (endpoints: reproduction and mortality). For springtails, the calculated NOEC was ≥ 500 mg/kg soil dw. [Staples et al., 2010] In a study, emergence and growth of 6 different terrestrial plant species were assessed after BPA exposure. The following toxicity values were estimated [EC, 2010]: Corn (Zea mays):

130 mg/kg soil dw (21-d NOEC)

Oats (Avena sativa):

47 mg/kg soil dw (21-d NOEC)

Wheat (Triticum aestivum):

47 mg/kg soil dw (21-d NOEC)

Cabbage (Brassica oleracea):

50 mg/kg soil dw (21-d NOEC)

Soybean (Glycine max):

320 mg/kg soil dw (21-d NOEC)

Tomato (Lycopersicon esculentum):

20 mg/kg soil dw (21-d NOEC)

Conclusion of the ecotoxicological risk assessment of the EC In 2008, in their PBT assessment of BPA in the risk assessment report, the EC concluded, that there are “no reliable chronic NOEC values below 0.01 mg/l, although there are some less reliable values and indications of possible effects at this level. Bisphenol-A has been shown to have effects on the endocrine systems of a number of organisms. It is therefore considered to meet the T criterion”. [EC, 2008a, p. 137] They noted that the exposure assessment is basing on data provided by the industry. Much of them is basing on sitespecific considerations. [EC, 2008a] Later study results (after 2008) suggest that there are no NOEC values below 0.01 mg/L. 28

3.3.2 Effects on wildlife Various studies had investigated the effects of BPA on wildlife. For a wide variety of species, effects have been shown generally at high BPA doses. Environmental systems are complex, exposure levels are varying spatially and trophic interactions occur. Thus, there are only a few studies available that investigated the effects of chemical exposure on in situ, and many studies have been performed in laboratory. [Flint et al., 2012] However, most studies show effects on the reproductive system and on development of wildlife animals.

Invertebrates and fish In studies on endocrine disruption, invertebrates are frequently used as bioindicators in laboratory and in situ, because of their sensitivity to BPA at environmentally relevant concentrations [Flint et al., 2012]. In all investigated invertebrates endocrine-signaling mechanisms exist, although the hormones that are involved in invertebrates are not always similar to the hormones in vertebrates. There is evidence for effects on development, fecundity and reproduction. [EEA, 2012] Commonly used species for toxicity tests in invertebrates and fish include insects, crustaceans, molluscs, annelids, isopods, and nematods, and in fish mainly Fathead Minnow (Pimephales promelas), Japanese Medaka (Oryzias latipes), Zebrafish (Danio rerio) and Rainbow Trout (Oncorhynchus mykiss). [Flint et al., 2012] Flint et al. reviewed several toxicity studies in invertebrates and fish. In various species, different effects were shown at various BPA doses. The reported effects are shown in Table 5. Further, Mihaich et al., 2009 investigated the acute and chronic toxicity to BPA in invertebrates and compared their results with other findings in the scientific literature. The EC50 values for acute toxicity ranged between 1.1 (Mysid shrimp, Mysidopsis bahia) and 10.2 mg/L (Water flea, Daphnia magna) and the NOEC between 0.51 and 4.1 mg/L, respectively in various species. Relating to the chronic toxicity, the authors reported several NOECs ranging between 0.025 (Snail, Marisa cornuarietis) and > 3.16 mg/L (Water flea, Daphnia magna) and LOECs ranging between 0.46 (Hydra, Hydra vulgaris) and 160 mg/L (Sponge, Heteromyenia sp.). [Mihaich et al., 2009] Those and many other studies have shown the effects of BPA on development and reproduction. However, many observed BPA levels are above concentrations relevant for environment. [Flint et al., 2012] Additionally, although well-researched areas in toxicology of BPA as well as other EDs in invertebrates exist, there are gaps in our knowledge.

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Further investigations and data are needed, especially on invertebrate biodiversity and abundance, especially in dependency to food chains of vertebrates. [EEA, 2012]

Table 5. Overview of reported effects in various toxicity studies in invertebrates and fish reviewed by Flint et al., 2012 Effect(s)

BPA exposure

Species

Reduced time to molt and altered sex ratios, reduced overall growth, identifiable protein expression changes, increased miscarriage rate and reduced reproductive allocation

10,000 µg/kg for 15 days to 16 weeks

Isopods

Developmental inhabitation and developmental stimulation

100 µg/L for 2 weeks as well as 0.1 µg/L for 4 weeks, and 12.5 µg/L for 3 weeks, respectively

Crustaceans

50 µg/L for 14.7 weeks

Crustaceans

228 µg/L for 48 h

Crustaceans

50 µg/L for 3 weeks

Molluscs

5 µg/L for 9 weeks, 0.19 µg/kg for 4 weeks

Molluscs

1 µg/L for 5 months

Molluscs

1,000 µg/L for 2 weeks

Carps

Vitellogenin induction

534 µg/L for 1 week 500 µg/L for 1 week 160 µg/L for 2 weeks 100 µg/L for 2 weeks 50 µg/L for 3 weeks 40 µg/L for 4 weeks 10 µg/L for 2 weeks

Zebrafish Rainbow Trout Fathead Minnows Atlantic Cods Carps Goldfish Seabass

Altered sex steroid levels

59 µg/L for 2 weeks

Turbot

Increased percentage of spermatocytes and reduced numbers of mature spermatozoa

16 µg/L for 164 days

Fathead Minnows

Decreased estrogen to androgen rations in blood

1-10 µg/L for 2 weeks

Carps

Complete inhibition of ovulation

5 µg/L for 3.5 months

Brown Trout

1.75 µg/L for 3.5 months

Brown Trout

1 µg/L for 164 days

Fathead Minnows

228 µg/L for 48 h

Zebrafish

42 µg/L for 6 weeks

Cnidarians

11.4 µg/L for 1 h

Annelids

300 µg/L for 0.5 h

Echinoderms

Crustaceans

Accelerated oocyte development and decreased number of offspring Suppressed development

Molluscs Spawning induction, ooxyte and ovarian follicle damage Increased female fecundity Superfeminisation, oviduct rupture and mortality Fish Decreased estrogen to androgen rations in blood

Reduced sperm quality and delayed ovulation Increased percentage of spermatocytes and reduced numbers of mature spermatozoa Feminized brains in embryos

Cnidarians Tentacle damage and contracted bodies Annelids Premature metamorphosis of larvae Echinoderms Fertilization reduction and increased larval deformities

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Reptiles Reptiles are not commonly used for studies on environmental contaminations and ecological risk assessments. [Campbell and Campbell, 2002] Stoker et al., 2003 investigated endocrine disruption caused by BPA in Broad-snouted Caiman (Caiman latirostris) after exposure in ovum and its effects to the reproductive system. The species has a temperature-dependent sex determination, which was amongst the gonadal histoarchitecture endpoint. The authors reported that BPA exposure leads to developmental effects, like sex reversal and altered gonadal histoarchitecture in embryos. The shown effects were similar to the effects after E2 exposure, but 100-fold lower. Additionally, they reported abnormal seminiferous tubles after a BPA exposure of 90 µg/egg (1400 µg/L). At BPA concentrations of 9 mg/egg (140,000 µg/L) during critical periods for gender determination, sex reversal from male to female in male-determining temperature incubated eggs was shown. [Stoker et al., 2003; Flint et al., 2012]

Amphibians Amphibians are a classical model species for investigations on endocrine disruptors and their effects on the reproductive and thyroid system. They are excellent bioindicators for exposure assessment in the aquatic phase. The amphibians’ egg and larval stages are most sensitive, because exposure is possible at critical periods of development. As juveniles and adults, the amphibians may have different life stages, from aquatic to terrestrial ones. Additionally, they have endocrine systems organised like those of most vertebrates. [Kloas and Lutz, 2006] Flint et al., 2012 reviewed several studies of BPA exposure to frogs. The reported effects are shown in Table 6. Goto et al., 2006 investigated the effects of BPA on 3,3’,5-triiodothyronine (T3)-induced and spontaneous anuran tadpole tail regression in Japanese Wrinkled Frog (Glandirana rugosa), Tropical Clawed Frog (Silurana tropicalis) and African Clawed Frog (Xenopus laevis). They reported that BPA regulates the gene expression mediated by thyroid hormone receptors down. [Goto et al., 2006] In another study in frogs, Heimeier et al., 2009 exposed growing tadpoles to 0.1 and 10 µM BPA. They reported that both BPA doses affected gene expression controlled by T3 hormone and metamorphosis. Thus, BPA inhibits the thyroid hormone, which directs the development. [Heimeier et al., 2009]

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Table 6. Overview of reported effects in various toxicity studies in frogs reviewed by Flint et al., 2012

Effect(s)

BPA exposure

Species

No vitellogenin induction

22,800 µg/L

European Common Frogs

Head malformations, scoliosis, and organogenesis suppression

5,700 µg/L

African Clawed Frogs

0.83-4,97 µg/L for 90 d

African Clawed Frogs

4,600 µg/L for 93 h

African Clawed Frogs

200 µg/L for 45 d

Dark Spotted Frogs

Female-biased sex ratio

22.8 g/L for 12 weeks

African Clawed Frogs

Sex reversal from male to female

22.8 µg/L for 2 weeks

African Clawed Frogs

Vitellogenin induction

22.8 µg/L for 36 h

African Clawed Frogs

Inhibited spontaneous metamorphosis

2.28 µg/L for 9 d

Western Clawed Frogs

No effects Abnormal gut coiling, edema, microcephaly, and decreased body length Tail flex malformations

Birds In birds, only a few studies estimating the effects of BPA are available at the current time. [Flint et al., 2012] Berg et al., 2001 assessed the effects of BPA on reproductive organs in avian embryos by injecting BPA into the yolk of quail and chicken eggs. Embryonic oviduct malformations were shown in female quail embryos as wells as feminization of the left testis in male chicken embryos at BPA concentrations of 200 µg/g egg in incubation. Mortality caused by BPA was shown in chicken embryos at 67-200 µg/g egg. [Berg et al., 2001] Furuya et al., 2002 investigated the BPA effects on growth of combs and testes of male chicken. They reported that oral BPA doses as low as 2 µg/kg bw administrated every 2 days for 23 weeks showed delayed growth of comb, wattle and testes in male chicken. No differences were found at oral BPA doses of 200,000 µg/kg bw once a week from 2-16 weeks of age between juvenile chickens and controls. [Furuya et al., 2002] Halldin et al., 2001 estimated the uptake and distribution of BPA in embryos and Japanese Laying Quail (Coturnix coturnix japonica) and variables related to the reproduction on adult quail after administration of BPA into the yolk of the eggs. They did not find any effects of BPA at concentrations of 67 and 200 µg/g egg. [Halldin et al., 2001]

Mammals Studies in mammalian wildlife are very rare. It is difficult to assess specific BPA effects in wildlife, because mammals are likely to experience lower BPA exposure levels than other 32

taxa. Further, BPA exposure levels may vary widely in dependence on exposure duration, food consumption and contaminated areas. Most studies in BPA effects on the mammalian wildlife rely on laboratory data at the current time. [Flint et al., 2012] Nieminen et al., 2002a investigated BPA effects on field voles (Microtus agrestis) in wildlife, and found increased plasma testosterone levels at BPA concentrations of 250 mg/kg/d. [Nieminen et al., 2002a] No BPA effects were shown in polecat (Mustela putorius) [Nieminen et al., 2002b]. They indicated that more studies on wildlife animals are needed.

Plants and algae In aquatic plants and diatoms the EC50 values were between 1.0 (Marine diatom, Skeletonema costatum) and 20 mg/L (Duckweed, Lemna gibba). The NOEC ranged between 0.4 (Marine diatom, Skeletonema costatum) and 7.8 mg/L (Duckweed, Lemna gibba). [Mihaich et al., 2009] In terrestrial plants the effects of BPA on emergence and growth were assessed according to the OECD Test Guideline 208. Investigated species were corn, oats, wheat, cabbage, soybean and tomato. The lowest EC25 value was estimated at 19 mg/kg dry weight in tomato plants. A NOEC value was derived of 20 mg/kg dry weight, which is also used in the derivations of the PNEC in the current European Risk Assessment Report. [EC, 2010]

Summary of wildlife effects Most of the studies investigating BPA exposure on wildlife show effects on reproduction and development. Invertebrates are a sensitive species to investigate BPA at environmentally relevant BPA doses. For example, a study has shown BPA effects in molluscs at a dose of 0.19 µg/kg (exposition for 4 weeks), which suggests effects at low doses. [Flint et al., 2012] Additionally in amphibians that are classical models for investigations of endocrine disrupting impacts of BPA effects where shown at low doses. [Kloas and Lutz, 2006] Reptiles also react on BPA exposure but in comparison to e.g. fish, invertebrates or amphibians effects where shown at higher doses of BPA [Stoker et al., 2003; Flint et al., 2012]

3.3.3 Toxicity values The Predicted No Effect Concentration (PNEC) is defined as the predicted concentration of a chemical derived from toxicity tests that does not cause any toxic effects. [Fent, 2007]

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Table 7 shows the derived PNECs for aqua, sediment, soil and secondary oral toxicity of BPA used for the risk assessment in the RAR of the EU.

Table 7. PNECs of BPA used for risk assessment in the European Risk Assessment Report [EC, 2010] PNECs in the EU RAR Aqua (freshwater)

1.6 µg/L

Aqua (marine water)

0.15 µg/L

Sediment (freshwater)

24 µg/kg ww (63 µg/kg dw)

Sediment (marine water)

2.4 µg/kg ww (6.3 µg/kg dw)

Soil

3.7 mg/kg dw

Oral

2.67 mg/kg food

Abbreviations: dw, dry weight; PNEC, predicted no effect concentrations; RAR, Risk Assessment Report; ww, wet weight.

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4 Human Health 4.1 Human exposure 4.1.1 Food and food contact materials Food cans and containers BPA is used as application of epoxy resins for inner can coatings and is able to leach from food cans and containers. Several studies have investigated BPA migration from food cans and containers. [Vandenberg et al., 2007] For example, Brotons et al., 2005 measured the BPA leaching from food cans that ranged between 4 – 23 µg BPA per can. [Brotons et al., 1995] Kang et al., 2003 investigated the relationship between BPA migration from cans and containers, heating time and temperature. In cans containing water samples more extensive effects were shown at constantly high temperatures than during the heating time. [Kang et al., 2003] In their study, Takao et al., 2002 estimated the relationship between the BPA migration levels and the BPA content of can linings, heating temperature and the lining material. They found associations between the BPA concentrations and the temperature of heat-treatment. [Takao et al., 2002] Goodson et al., 2004 performed an experiment to investigate the BPA migration into food under different storage conditions and levels of can damage. They reported that 80 to 100% of the BPA in the coating migrated into food directly after processing. These concentrations did not change by extended storage or can damage. In contrast to other studies, they did not find any differences in BPA concentrations before and after heating. [Goodson et al., 2004] Plastic food contact materials BPA is used for PC manufacture and as additive in PVC products (cf. Chapter 2.3), and is able to migrate from plastic food contact materials into food. Several studies have estimated the migration of BPA from plastic materials. Lopez-Cervantes and PaseiroLosada, 2003 measured the BPA content of 5 PVC-stretch films used for food packaging and its migration into water, acetic acid and olive oil after storage over 10 days at 40°C. The BPA contents in the stretch films ranged between 43 and 483 mg/kg in 4 stretch films (BPA was not detected in one stretch film), and in olive oil the highest BPA migration was shown. The measured concentrations of BPA were lower than the specific migration limit (SML) of the EU, but the authors concluded that PVC-stretch films nevertheless may have a significant contribution to BPA contamination of food. [López-Cervantes and PaseiroLosada, 2003] In their study, Kubwabo et al., 2009 performed a migration test using several plastic containers: PC and non-PC baby bottles, baby bottle liners and reusable 35

PC drinking bottles. They stimulated the BPA migration into aqueous, acidic, alcoholic and fatty foods. The detected BPA migration ranged between 0.11 µg/L (at 40°C in water incubated for 8 h), and 2.39 µg/L (at 40°C in 50% ethanol incubated for 240 h). An increase of the BPA leaching from PC bottles was shown with increased temperature and incubation time. The authors concluded that glass baby bottles, and plastic bottles made of polysulfone, polypropylene and polystyrene are reasonable alternatives. Additionally, the use of reusable drinking bottles for cold beverages and short times would result in lower BPA migrations rates. [Kubwabo et al., 2009] Paper food contact materials and cardboards BPA is also used in the paper production. Thus, it may be present in papers with food contact. A study investigating kitchen paper towels has shown that virgin papers did not contain any BPA. Kitchen towels made from recycled papers have shown BPA ranges between 0.55 and 24.1 mg/kg. [Vandenberg et al., 2007] Ozaki et al., 2004 investigated virgin and recycled paper products that are used as food contact materials. 67% of recycled paper products contained BPA ranged from 0.19 to 26 µg/g. In virgin papers, BPA was also detected, but the measured levels were ≥ 10-fold lower than in the recycled paper products. [Ozaki et al., 2004] Those and several other studies indicated that papers and cardboards used for food contact are potential sources of BPA contamination. [Vandenberg et al., 2007] Human exposure via food Through migration of BPA from food contact materials like plastic products, cans, paper products and cardboards into food, humans are exposed to BPA. Many studies have investigated the BPA content of various foods. For example, Cao et al., 2011 analysed 154 food composite samples from a total diet study in Quebec City, Canada. They detected BPA in 36% of the tested samples. BPA was found in high concentrations in canned food samples (maximum: 106 ng/g in canned fish). In not-canned food, they found BPA in yeast, baking powder, some cheeses, some cereals and breads, as well as in fast food. (maximum: 8.52 ng/g in yeast). Additionally, they investigated the dietary intake of BPA to humans, and concluded that the intakes are low for all age- and sex-groups. [Cao et al., 2011] Rudel et al., 2011 performed a dietary intervention study with 20 participants from families from California, USA. They measured the urinary BPA levels before, during and after a dietary intervention with fresh food. The authors reported a significant decrease of BPA levels of 66% in urine during the fresh food intervention period. [Rudel et al., 2011]

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The findings in many studies suggest that food, especially canned food, is an important source of BPA exposure of humans.

4.1.2 Air and house dust Another source of BPA exposure is house dust. Through production and use, BPA is able to leach into the environment and can be measured in air and house dust. [Geens et al., 2009] Rudel et al., 2003 measured BPA levels in 120 indoor house dust samples. They found BPA concentrations between 0.2 and 17.6 g/kg in 86% of all samples. [Rudel et al., 2003; Vandenberg et al., 2007] BPA levels were measured in an additional study in outdoor air in Japan at mean levels of 0.51 ng/m3. They found increasing BPA levels from autumn to winter and decreasing BPA levels from winter to spring. Thus, mild seasonal variations were reported. [Vandenberg et al., 2007] In another study, the human exposure of BPA through indoor dust intake in Belgium was assessed in 18 houses and 2 offices. They found a mean BPA concentration of 2001 ng/g in houses, and a mean concentration of 6530 ng/g in the offices. Thus, in the offices, the BPA levels were much higher than in the house dust samples. Possible contamination sources in the offices may be office-related applications (e.g. electronic and electrical equipment and office furniture). [Geens et al., 2009]

4.1.3 Dental products In dentistry, various resin-based monomers are used as adhesives, preventative sealants and restorative materials. BPA diglycidyl methacrylate is synthesised from BPA and has been used since the 1960’s in a multitude of dental materials. It has been shown that some quantities of the unpolymerised monomers leach from dental materials. [Vandenberg et al, 2007] During the dental filling with sealants and composite resins, compounds congeal by polymerization before their use. Studies have shown that unpolymerised monomers may be dissolved in salvia. [Sasaki et al., 2005; Rathee et al., 2012] Sasaki et al., 2005 investigated changes in BPA levels in saliva samples of 21 patients after restoration with composite resins from 9 manufactures with ELISA. They measured BPA at several 10 to 100 ng/ml salvia following treatment with composite resins. [Sasaki et al., 2005] The findings of several studies suggest that dental products are also a source for BPA exposure, because of the leaching of small amounts of BPA from dental products immediately after the application to the teeth. [Rathee et al., 2012]

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4.2 Metabolism and Toxicokinetics After oral administration, BPA undergoes a rapid adsorption from the gastrointestinal tract (GIT) into the human body. In the liver, BPA is conjugated with glucuronic acid to BPAglucuronide (BPA-G), which is the main metabolite of BPA. The glucuronidation rate of BPA is limited by the transport rate from the GIT into blood and liver. Hepatocytes and microsomes from the liver and the intestine are catalysing the reactions. Other metabolites are formed after saturation of the pathway of glucuronidation only after exposure to of higher BPA doses. After oral administration, BPA is eliminated within 24 hours via urine. The BPA metabolism occur in a different way in rodents than in humans: whereas BPA follows the enterohepatic circulation, in humans this does not occur. [Völkel et al., 2002; Doerge and Fisher, 2010] The toxicokinetics and the metabolism of BPA at orally administered low doses were examined by Völkel et al. (2002). They administered 5 mg D16-BPA (0.054-0.090 mg/kg bw), which was a 10-fold higher dose than the estimated human exposure level of 0.6 mg/day, to volunteers and estimated the BPA levels in urine as well as in blood samples. They reported a half-time for D16-BPA-glucuronide for the urinary elimination of 5.4 h, and a terminal half-time for clearance from the blood of 5.3 h. After 24 to 34 h the concentrations were below the limit of detection (LOD). D16-BPA-glucuronide has an urinary recovery rate of 100%. Free D16-BPA-glucuronide in blood was not detected, which supported that BPA is adsorbed rapidly from the GIT, is conjugated with glucuronic acid in the liver and eliminated rapidly as glucuronide via urine. Additionally, this suggests, that BPA does not pass the enterohepatic circulation in contrast to rodents. [Völkel et al., 2002; Chapin et al., 2008]

4.3 Toxicity 4.3.1 Acute Toxicity In their Risk Assessment Report (RAR), the European Union reported that no useful information and adequate studies on the acute toxicity of BPA in humans are available. They reviewed data from some studies in animals and concluded that these data indicate a low acute toxicity of BPA by all routes of exposure (oral, dermal or via inhalation) that are relevant for humans. [EC, 2010]

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4.3.2 Irritation and Sensitisation According to the CLP regulation (Annex I), BPA can cause serious eye damage, skin sensitisation and respiratory irritation. [ECHA, n.r.g]

4.3.3 Repeated Dose Toxicity The EU reported in their RAR that no adequate information about repeated dose toxicity data for BPA in humans is available. But several studies had investigated repeated BPA exposure in animals. [EC, 2010] Inhalation: Inhalation studies in rats showed the same effects for repeated exposure than for a single exposure (slight upper respiratory tract epithelium inflammation). In a 13-week study in rats a NOAEL was identified of 10 mg/m3. [EC, 2010] Oral: In a two-generation reproduction toxicity study in CD-1 mice it was shown, that the repeated toxicity of BPA showed effects on bodyweight gain, liver and kidney. The identified NOAEL was 50 mg/kg/d. They concluded that BPA is not considered to be a reproductive or developmental toxicant in mice. [Tyl et al., 2008] However, several other studies have shown effects on reproductive system and development. Therefore, there is some concern of effects of BPA to reproduction and development (see Chapter 4.3.7). Dermal: For dermal repeated exposure to BPA, there is no animal data available. [EC, 2010]

4.3.4 Estrogenicity Estrogenic activity The first identification of the estrogenicity of BPA was in 1936 by Dodds and Lawson. [Dodds and Lawson, 1936] Estrogen is a steroid hormone that has important influences on growth, differentiation and function of many tissues, including those of the male and female reproductive systems (uterus, ovaries, vagina, mammary gland, testes, prostate and epididymis). It is also very important for the central nervous system, cardiovascular system and the bone maintenance. In target body cells, the estrogen binds to the estrogen receptors ERα and ERβ with a high specificity and affinity. After the binding, the ER changes its conformation and is able to interact with chromatin. Because of this, the ER modulates the transcription of target genes. BPA has the ability to bind to these ERs, but its affinity is 10,000-fold weaker than that of estrogen. The relative transactivation activity for ERα and ERβ is about 50 and 41, whereas the transactivation activity of the estrogen 17β-estradiol is set to 100. [Kuiper et al., 1998] Additionally, several in vitro studies have 39

shown interactions of BPA with other receptors like membrane bound ER and estrogenrelated receptor γ (EERγ). [FDA, 2008] The main BPA metabolite BPA-G has not shown a significant estrogen activity in in-vitro or in-vivo test systems. Thus, the detection and estimation of free unconjugated BPA levels in target tissues at sensitive stages of live of the test species is important for the BPA safety analysis. In conclusion, free BPA has an estrogen activity, but its main metabolite BPA-G has not. Data of several studies indicate that some free unconjugated BPA is able to enter different tissues. In the tissues, BPA may have an extended resistance time in its free form. Additionally, BPA is able to enter fetuses via the placenta, either in animals or in humans. However, there are many uncertainties (see Chapter 5.8) [FDA, 2008] There is a lack in understanding some of the pathways of BPA in the human body, as well as how the estrogenic activity of BPA occurs relating to its fast elimination. Further scientific examinations are needed.

Estrogenicity Various studies had estimated the estrogen potency of BPA and there is a considerable variability in these study results. One explanation for the wide range of reported values in different studies is the difference between laboratories. Another explanation is the difference in defining the relative estrogen potency of BPA in some assays. [Chapin et al., 2008] For example, Kurosawa et al., 2002 performed a luciferase assay on 3 independent cell lines from different tissues using ERα- and ERβ-reporting systems to characterize the estrogenic effect of BPA. They found estrogenic activities in all cell lines via ERα and ERβ. Their results indicated that BPA acts both as an agonist and an antagonist in some cell types via ERα, whereas it only acts as an agonist of estrogen via ERβ. [Kurosawa et al., 2002] By contrast, Rajapaske et al., 2001 performed a study using a recombinant yeast assay for ERα activation and found additive effects of BPA and 17β-estradiol. [Chapin et al., 2008; Rajapaske et al., 2001] Compared to those results, Suzuki et al., 2001 investigated the effects of BPA on the cellular proliferation of human breast cancer cells by a modified E-screen assay and found synergistic effects of BPA and 17β-estradiol. [Suzuki et al., 2001] Estrogenic activity has not been found for BPA-glucuronide or -sulfate. [Chapin et al., 2008] Nevertheless, BPA has a weak estrogenic activity that can lead to various human health effects by stimulating cell responses and various molecular endpoints even at low doses. [Vandenberg et al., 2007; Newbold et al., 2009] BPA may have effects on paraovarian cysts, ovarian lesions and stromal polyps in uterus [Newbold et al., 2009], may have an 40

impact on estrogen cycles, premature start of puberty, increase of body weight and fertility problems as well as effect on reproduction and development. [Vandenberg et al., 2007]

4.3.5 Mutagenicity The EU reviewed in their RAR various studies that investigated possible mutagenic effects of BPA. BPA was demonstrated to have aneugenic potential and micronuclei formation invitro without metabolic activation. In other studies disruption of microtubule formation and presence of DNA adducts were demonstrated. In those reviewed studies no evidence of gene mutations and chromosomal aberrations in-vitro were shown. [Chapin et al., 2008]

4.3.6 Carcinogenicity For humans, there are no epidemiological data of the carcinogenicity of BPA available. [Chapin et al., 2008] NTP, 1982 investigated the possible carcinogenicity in mice and rats. BPA was administrated orally in different concentrations. They reported an increase of some cancer types in rats, but concluded that none of them was associated in a clear way with BPA exposure. The European Union reviewed this study and concluded that the increase of the tumors (leukemia, mammary gland fibroadenoma and Leyding cell tumors) had no toxicological relevance. Thus, BPA has no carcinogenic potential. [NTP, 1982; Chapin et al., 2008] Additionally, the International Agency for Research on Cancer (IARC) and the U.S. EPA made a weight-of-evidence approach in 2002 and concluded that BPA has no carcinogenic potential to humans. [Haighton et al., 2002] Also in the Expert Meeting of the FAO and WHO in 2010 it was concluded that currently there is no evidence for a carcinogenicity of BPA. [Bucher, 2010] Keri et al., 2007 evaluated the evidence of the carcinogenic activity of BPA. Based on the existing evidence the authors were confident that natural 17β-estradiol is a carcinogen. Additionally BPA acts as an ED with some estrogenic properties. It is likely that BPA may be associated with increased prevalence of cancer of the hematopoietic system and with significantly increased prevalence in cell tumors in testes, that BPA alters microtubule function, and that early life BPA exposure may induce pre-neoplastic lesions of mammary and prostate gland in adults lifetimes. It is furthermore likely that prenatal BPA exposure to environmentally relevant doses alters mammary gland development in mice and increases endpoints that are markers of breast cancer risk. Thus, it may be possible that BPA may 41

induce cellular transformation in vitro and may be able to promote tumor progression and to reduce the time to recurrence. [Keri et al., 2007]

4.3.7 Reproductive and Developmental Toxicity Studies in humans In literature, a few studies of birth outcomes in humans and BPA exposure are available. Padmanabhan et al., 2008 investigated in a cross-sectional study maternal BPA levels in blood from 40 women in USA in a hospital and their possible relationship to gestational length and birth weight of their offspring. They found no correlations between BPA levels and the gestational length or birth weight of the offspring. [Padmanabhan et al., 2008; Golub et al., 2009] Wolff et al., 2008 compared in their study maternal BPA levels in women in the 3rd trimester of pregnancy to birth weight, length, head circumference and gestational age of their offspring. They did not find any effects of BPA in their study. [Wolff et al., 2008; Golub et al., 2009] Hanaoka et al., 2002 investigated possible correlations between BPA exposure and hormone levels in workers in Japan that were exposed to an epoxy hardening agent, which consists of a mixture containing, amongst others, BPA diglycidyl ether (10-30%). They reported that the measured BPA concentrations in urine were significantly higher in the exposed workers than in the control group. Additionally urinary organic solvent metabolites were detected more frequently in samples of the workers. No differences were found in plasma testosterone or LH concentrations. For the exposed workers, the plasma FSH (follicle-stimulating hormone) concentrations were significantly lower than in controls. Significant associations between FSH and BPA following adjustment for alcohol intake were observed. The authors concluded that BPA may disrupt gonadotropic hormone secretion in males. [Hanaoka et al., 2002] Takeuchi et al., 2004 investigated BPA concentrations in serum in women with ovarian dysfunction and obesity. They obtained serum samples from obese and non-obese women with normal menstrual cycles, from patients showing hyperprolactinemia and from patients showing hypothalamic amenorrhea, and obese and non-obese women with polycystic ovary syndrome (PCOS). In obese and non-obese women with PCOS and obese normal women, the measured BPA concentrations in serum were significantly higher than in nonobese normal women. In all subjects, significant positive correlations between BPA concentrations in serum and concentrations of total testosterone, free testosterone, androstenedione and dehydroepiandrosterone sulfate were found. These findings showed

42

a strong relationship between BPA levels in serum and androgen concentrations. [Takeuchi et al., 2004] Studies in laboratory animals Kim et al., 2001 investigated possible adverse effects of BPA to pregnancy and embryofetal development after oral maternal BPA exposure during pregnancy in SpragueDawley rats. The administered BPA dose levels were 0, 100, 300 and 1000 mg BPA/kg bw. In the high-dose group, female rats showed an increased pregnancy failure, reduced maternal body weights (also shown in the mid-dose group), expansion and congestion of stomach and intestines, decreased body weights of male fetuses (also shown at mid-dose) and of female fetuses, increased fetal death, early resorption, reduced anogenital distance in males (also shown in the mid-dose group) and reduced ossification. Dysmorphogenesis was not shown. This study supports that prenatal BPA exposure has effects on the developmental toxicity in rats. [Kim et al., 2001; Chapin et al., 2008] Ema et al., 2001 performed a two-generation reproduction study in rats to investigate lowdose effects of BPA. They administrated BPA doses between 0.2 and 200 µg/kg/d by gastric intubation to male and female rats. They did not find any significant BPA related effects in reproductive or developmental parameters in rats. [Ema et al., 2001] Tyl et al., 2008 performed a two-generation reproductive toxicity study in CD-1 (Swiss) mice by administration of different BPA concentrations (0 to 600 mg BPA/kg/d). They reported that no BPA related effects were shown on adult mating, fertility, gestational indices, ovarian follicle counts, offspring sex ratios, offspring postnatal survival, sperm parameters or weight of reproductive organs or histopathology, estrous cyclicity or precoital interval. BPA concentrations of 600 mg BPA/kg/d in the adult mice reduced body weights, increased liver and kidney weights. Centrilobular hepatotrophy and renal nephropathy in male mice were shown. At the same concentrations, BPA reduced F1/F2 weaning body weights, weanling spleen and testes weights. The authors reported a systemic NOEL of 5 mg/kg/d and a reproductive/developmental NOEL of 50 mg/kg/d. Thus, they concluded that BPA is not considered to have reproductive or developmental toxicity in mice. [Tyl et al., 2008] Newbold et al., 2009 investigated the BPA effects in pregnant CD-1 mice that were treated with BPA on days 9 to 16 of gestation with concentrations between 0.1 - 1000 µg/kg/d. The reproductive tissues of their offspring were evaluated at the age of 18 months. The authors reported a significant increase of ovarian cysts in the 1 µg/kg BPA group. They observed an increase in progressive proliferative lesions after BPA treatment. Uterus metaplasia, vaginal adenosis, atypical hyperplasia and stromal polyps of the uterus were shown, 43

although these findings were not statistically different from the control group. The authors concluded, that the data suggest long-term adverse reproductive and carcinogenic effects after BPA exposure during critical periods of differentiation. [Newbold et al., 2009] Negative effects on fertility and spermatogenesis of male rodent offspring were shown after maternal BPA exposure of environmentally relevant BPA doses. Perinatal BPA exposure showed effects on the male germ line. This was manifested as impairments in the fertility of the male offspring. [Salian et al., 2011] Richter et al., 2007 reviewed several studies that investigated in-vivo effects of low doses (below 50 mg/kg/d) of BPA in laboratory rodents. A multitude of the shown BPA effects were similar to the effects of the estrogens diethylstilbestrol and ethinylestradiol in models, but the potency of BPA is 10 to 1000-fold lower. The authors reported that BPA exposure to adult male rodents affect the reproduction tract. In the long term, the developmental BPA exposure leads to effects in brain and male reproductive system, and to metabolic processes. Further confirmation is needed, but there are indications, that BPA exposure in adults affects the female reproductive system, immune system, and developmental effects in the female reproductive system. [Richter et al., 2007] Summary of reproductive and developmental toxicity of BPA Many studies have been performed to investigate reproductive and/or developmental effects of BPA. Only a few of them are described in this thesis. The NTP-CERHR Expert Panel (Chapin et al., 2008) reviewed a multitude of the available studies. Data on effects of BPA to human development are not available. Additionally, there are insufficient data on reproductive effects in humans. However, a large number of experimental animal studies are available. Concerning the developmental toxicity of BPA, from rodent studies the Expert Panel concluded that BPA does not cause malformations and/or birth defects at levels up to 640 mg BPA/kg/d in rats and 1250 mg BPA/kg/d in mice, does not alter fertility after gestational exposure up to 450 mg BPA/kg/d in male and female rats and 600 mg BPA/kg/d in male and female mice, does not affect prostate weight permanently at BPA doses up to 475 mg/kg/d in adult rats and 600 mg/kg/d in mice, does not cause prostate cancer up to BPA doses up to 158 mg/kg/d in adult rats and 600 mg/kg/d in adult mice, and does not change puberty age in male and female rats at high doses of ca. 475 mg BPA/kg/d. However, the rodent studies suggest that BPA cause behavioral and neural alterations in relation to disruptions in normal sex differences at concentrations of 0.01 to 0.2 mg BPA/kg/d. Concerning the reproductive toxicity, the authors reported that in rodents there is sufficient evidence that BPA causes reproductive 44

toxicity with chronic or subchronic oral BPA exposure (with a LOAEL of ≥ 475 mg/kg bw/d and a NOAEL of 47.5 mg/kg bw/d). After the review of many studies, for pregnant women and fetuses the NTP-CERHR Expert Panel has some concern for neural and behavioral effects, minimal concern for prostate effects and potential effects of accelerated puberty, and negligible concern for birth defects and malformations. For infants and children the Expert Panel has some concern for neural and behavioral effects and minimal concern for effects of accelerated puberty. For adults it has negligible concern for adverse reproductive effects. [Chapin et al., 2008] Studies published after the NTP-CERHR Expert Panel Report showed different results. While Tyl et al., 2008 did not consider that BPA has developmental or reproductive toxicity in mice [Tyl et al., 2008], findings by Newbold et al., 2009 suggest long-term adverse carcinogenic and reproductive effects at critical periods of differentiation in mice [Newbold et al., 2009]. Additionally, in a study performed by Salian et al., 2011 negative effects of BPA on fertility and spermatogenesis after maternal exposure in male rodent offspring have been shown, as well as effects on a male germ line [Salian et al., 2011].

4.3.8 Conclusions of the EC in their Risk Assessment Report The risks and health hazards related to BPA were evaluated in Europe under the Existing Substances Regulations (ESR) according to Directive 793/93/EEC and published by the EC in their BPA Risk Assessment Report. It was concluded that: -

there is a risk for workers and therefore, there is a need for risk reduction from BPA exposure because of the repeated dose toxicity of BPA including effects on respiratory tract, body weight gain and kidney, effects on liver and reproductive toxicity for manufacture of BPA and of epoxy resins, as well as skin sensitization at high BPA concentrations,

-

there are no health effects for consumers. Thus no further risk management activity is required under REACH,

-

there are no health effects for humans via the environment, and thus no further management activity is required, too,

-

there are no health concerns for combined exposure, and therefore no further management activity under REACH is required.

45

In the EU RAR, respiratory tract irritation, skin sensitization, repeat dose toxicity and reproductive toxicity have been identified as endpoints with concerns. Table 8 shows the identified dose descriptors for those endpoints [United Kingdom, 2008]: Table 8. Dose descriptors identified in the RAR for endpoints of concern [United Kingdom, 2008, p. 11]

4.3.9 Summary of the toxicity of Bisphenol A The acute toxicity of BPA is low. It can cause serious eye damage, skin sensitization and respiratory irritation. According to the CLP regulation, BPA is classified as Skin Sens. 1 (“may cause an allergic skin reaction”), Eye Dam. 1 (“causes serious eye damage”) and STOT SE 3 (“may cause respiratory irritation”), as well as Repr. 2. Therefore, BPA is suspected to damage fertility of the unborn child. [EC, 2010; ECHA, n.r.g] BPA is a weak estrogen and is able to bind at estrogen receptors. Thus, BPA can stimulate cell responses and molecular endpoints, even at low doses. [Kurosawa et al., 2002; Vandenberg et al., 2007; Newbold et al., 2009]. According to the RAR of the EU, there is no evidence of mutagenicity or carcinogenicity of BPA. [NTP, 1982; Chapin et al., 2008] However, some studies indicated that BPA may promote tumor progression. [Kari et al., 2007] There is some concern to the possible reproductive and developmental toxicity of BPA, but further studies are needed. [Chapin et al., 2008]

46

5 Chemical Policy 5.1 Former Chemicals Legislation in the EU In the EU, the former chemicals legislation contained a multitude of different regulations and directives that distinguish amongst others between so-called “new” and “existing” chemicals. This differentiation was introduced under the EC-regulation 793/93 and had its “cut-off date” in 1981: Chemicals on the market of the European Union that were listed in the European Inventory of Existing Commercial Chemical Substances (EINECS) before September 18th, 1981 were classified as “existing” chemicals and amounted > 100,000 various substances. The contemplated “new” chemicals were introduced to the market after 1981 and amounted > 3,800 different substances. Before the placement of these “new” chemicals on the market in amounts of more than 10 kg they have to be tested regarding their hazard for human health and environment. Such a regulation did not exist for “existing” chemicals, but several pieces of information on properties and usage were available. However, there is a lack of sufficient information. The responsibility of the implementation of risk assessments was on public authorities, not on the industry that produce, import and/or uses these substances. Because of this, the risk assessment was inert. The allocation of the responsibilities of the pre-REACH process was not appropriate, because the public authorities had the responsibility for undertaking risk assessments, rather than the industry that produced, imported or used the substances. This lead to the identification of only 141 substances produced in high volumes as priority substances for the risk assessment since 1993. A recommendation was only available for a small number of those chemicals for which a complete evaluation process was done under the ECRegulation 793/93. Additionally, information on usage of substances was difficult to obtain from industry. Furthermore, the pre-REACH process was very slow; thus, the marketing and use of only 100 chemicals was restricted since 1976. [EC, 2007b] The development and adoption of REACH was very important because of the large number of chemical substances produced and placed on the European market and because of the lack of adequate information on the hazard properties to human health and the environment [EC, 2012f].

Future Chemical Strategy In 2001, the EC published their Strategy for a Future Chemicals Policy. They reviewed the pre-REACH system and took this review as a basis for new strategies. Aims are the protection of human health and the environment, high levels of chemical safety, a 47

competitive chemicals industry, the prevention of fragmentation of the market of the EU, an increased transparency, the promotion of non-animal testing, conformity with international obligations under the WTO, and integration with international efforts. The overriding goal is the sustainable development. [EC, 2007a; EC, 2007b]

5.2 REACH Regulation REACH (Registration, Evaluation, Authorisation and Restriction of Chemical substances) is a Regulation of the European Community (Regulation (EC) No 1907/2006) that regulates chemicals and their safe use. This regulation was published in 2006 and entered into force in June 2007. It harmonizes the European law of chemicals and is valid for all member states in the EU. The aims of REACH are to grant a high level of protection of human health and the environment against chemical risks and a better and faster identification of the properties of chemicals. REACH depends on the responsibility of the industry to provide information on chemicals and their risk management. [EC, 2012f] Because of the production and placement on the market, the industry has certain knowledge of the chemicals and their potential risks. [EC, 2007b] Central point in the REACH system is the European Chemicals Agency (ECHA) located in Helsinki, Finland which manages the databases for operating the system. Further responsibilities of the ECHA are the coordination of the evaluation of suspicious chemical substances, and building up the database to make hazard information of these chemicals available for consumers and professionals. [EC, 2012f] Under REACH, the previously so-called “existing” and “new” substances are now described as “non-phase-in” and “phase-in” substances [EC, 2007b]: Phase-in substances are substances which fulfill one or more of the following criteria according to Article 2(20) of the REACH-regulation: -

Substances that are listed in EINECS

-

Substances manufactured in the EU but not put on the market of the EU after the 1st of June 1992

-

So-called “no-longer polymer” substances [ECHA, 2008]

48

Non-phase-in substances are substances which were not manufactured or placed on the EU market before the entry into force of REACH [EC, 2007b] and do not fulfill any criteria for the phase-in substances [ECHA, n.r.a].

5.2.1 Scope The European Commission describes the scope of REACH as the follows: “REACH is very wide in its scope covering all substances whether manufactured, imported, used as intermediates or placed on the market, either on their own, in preparations or in articles, unless they are radioactive, subject to customs supervision, or are non-isolated intermediates. Waste is specifically exempted. Food that meets the definition of a substance, on its own or in a preparation, will be subject to REACH however, such substances are largely exempted from Registration, Evaluation and Authorisation. Member States may exempt substances used in the interests of defence. Other substances are exempted from parts of REACH, where other equivalent legislation applies. The Commission will review the scope of the Regulation five years after entry into force” [EC, 2007b, p. 6]

5.2.2 The REACH process Table 9 gives an overview of the process of registration, evaluation, authorization and restriction of chemicals:

Table 9. Overview of the REACH process [cf. Schöning, 2010] Pre-registration Data sharing Registration Evaluation (dossier and substance evaluation) Authorisation Restriction Classification and Labelling

Gathering of information and ensuring responsibility and risk management by the industry

Controlling and request for further information by the ECHA and Member States

With support of the ECHA and the Member States, the Commission applies risk management systems

49

Pre-registration For manufacturers and importers of phase-in substances in quantities of ≥ 1 tonnes/year and manufacturers or importers of articles containing one or more substances with planned releases in quantities of ≥ 1 tonnes/year, the possibility was given to inform the ECHA about the substances they planned to register under REACH. This REACH process was called pre-registration and took place between 1st of June and 1st of December 2008. In case of pre-registration the companies had prolonged deadlines for the registration of their substances, whereas without pre-registration the companies have to register their substances before their production or their placement on market. [ECHA, 2008]

Registration Manufacturers and importers have to provide a registration dossier to the Agency for each substance which is produced or imported in quantities of ≥ 1 tonnes/year. In case of failure of the registration the company is not allowed to produce or import this substance any longer. The regulation excludes substances that are regulated under other legislations (e.g. medical products), or have very low risks (e.g. water, oxygen), or occur in nature (e.g. minerals, ores) as long as they are not modified, and polymers. However in certain cases the polymer’s monomers have to be registered. Annex IV and Annex V of the REACHregulation contain those exclusions of substances (a summary of all REACH Annexes is given in Annex A of this thesis). [EC, 2007b]

Phase-in substances have to be provided at following registration deadlines (Figure 8):  November 30, 2010: Registration of substances produced or imported above 1.000 tonnes/year above 100 tonnes/year when the substance is dangerous to aquatic organisms or the environment above 1 tonne/year or more when the substance is carcinogenic, mutagenic or toxic to reproduction (CMR)  May 31, 2013: Registration of substances produced or imported at 100 to 1000 tonnes/year  May 31, 2018: Registration of substances produced or imported at 1 to 100 tonnes/year

50

Non-phase-in substances were not produced or placed on market before June 1, 2008 (entry of force of the REACH regulation), unless they were notified under the EC Directive 67/548. Substances notified under this directive are considered to be registered under REACH. Manufactures or importers have to register the non-phase-in substance prior to production or import. [ECHA, n.r.a]

Figure 8. Registration deadlines [EC, 2007b]

According to the tonnage, REACH requires more or less information about the properties of the substance(s) from the manufacturers and importers. In Annexes VI to XI of the REACH regulation, information requirements and adaptations are set out. Manufactures and importers have to submit information on the substance identity, the physico-chemical properties, the mammalian toxicity and ecotoxicity, the environmental fate as well as information about manufacturing and usage and risk management measures. Additionally, manufactures or importers who register a substance in amounts of 10 or more tonnes per year have to conduct a Chemical Safety Assessment, which is a step-process: Collection and generation of information on intrinsic substance properties (step 1) Human health hazard assessment (step 2) physico-chemical hazard assessment (step 3) The environmental hazard assessment (step 4) PBT (persistent, bioaccumulative, toxic) and vPvB (very persistent, very bioaccumulative) assessment (step 5) 51

Exposure assessment (step 6) Risk characterisation (step 7) Step 6 and 7 are only needed, if steps 1-5 concluded that the substance is hazardous. The so-called Chemical Safety Report (CSR) contains the results of the Chemical Safety Assessment and is part of the registration dossier. [ECHA, n.r.b]

Registration of Monomers and Polymers In the REACH Regulation, Article 3(6), a monomer is defined as a substance that can form covalent bonds with a sequence of additional molecules to react to polymers (polymerisation reaction). The same substance is not regarded as a monomer for applications outside the scope of polymerisation. Any substance which is used as monomer for polymer production is by definition an intermediate (see “Registration of Intermediates”). Polymers are substances that consist of molecules that are characterised by sequences of one or more types of monomers. According to REACH Article 3(5) a polymer has to contain a sequence of 3 or more covalently bound monomer units. [ECHA, 2012h] Manufacturers or importers of a polymer have to submit a registration to the ECHA for the monomer(s) or any other substance(s) that has not been registered already by an actor further up the supply chain, if the polymer consists of ≥ 2% w/w of (a) monomer substance(s) and if the total quantity of the monomer(s) amount > 1 tonne per year. Polymers themselves are exempted from registration. [EU, 2006; ECHA, 2012h] The registration of the monomer substance or any other substance that is chemically bound to the polymer is not required if it has been already registered. [ECHA, 2012h]

Registration of Intermediates For intermediates, specific REACH registration requirements exist because of their special nature. [EU, 2006] According REACH, intermediates are defined as: Non-isolated intermediates: substances that are produced solely for transformation into other substances and are used up within this transformation reaction. They are exempted from REACH registration. 52

Isolated intermediates: all intermediates other than non-isolated intermediates. They can be registered reduced requirements and divided into two groups: o

On-site isolated intermediates: substances that are produced or used for chemical processing to become transformed into other substances. Their synthesis occurs on the same site, and they are operated by one or more legal entities.

o

Transported isolated intermediates: substances (e.g. pharmaceutical intermediates) that are produced or used for chemical processing to become transformed into other substances. The synthesis is transported between or supplied to other sites.

Prior to the registration of a substance it is important for the industry to check if a substance is an intermediate or not. The registration of intermediates is cheaper than a full registration. [CIRS, 2008-2012]

Evaluation The information of the manufacturers and importers is evaluated by ECHA and the member states of the EU to investigate their quality of the testing methods and registration dossiers. Actors in the evaluation process: Registrant (natural or legal person who produce or import a substance in amounts of 1 tonne or more per year and is able to act as registrant) Third parties (organisations, other companies, citizens, academics, etc. who e.g. provide information of testing proposal) ECHA (Secretariat - Committees - Member State Committee) Member States (are able to comment/propose amendments to draft decisions of ECHA, are able to propose substances for evaluation, impose national actions on substances, etc.) European Commission [ECHA, n.r.c]

53

Types of evaluation: Dossier evaluation: Examination of the registration dossier quality by ECHA o

Compliance check: Examination of the compliance of the registration dossier with the REACH-regulation requirements, whereas at least 5% of the registration dossiers have to be examined of each tonnage band

o

Testing proposals check: Examination of the test conditions before the test performing for the prevention of unnecessary animal tests

Substance evaluation: ECHA clears in cooperation with the member states suspicious risks to human health or to the environment and may require further information from the industry. After finishing the evaluation, further information on the substance may be required from the registrants. [EC, 2007b; ECHA, n.r.c]

Authorisation Aims of the authorisation of substances are to ensure the controlling of the risks of Substances of Very High Concern (SVHC) and the successive replacement of these substances by alternatives. Identification of SVHCs: Substances classified as CMR substances: carcinogenic, mutagenic and toxic for reproduction category 1A or 1B (according to Regulation No. 1272/2008) Substances classified as PBT or vPvB substances (according to REACH Annex XIII) Substances of equivalent concern as those above identified case-by-case; scientific evidence for the cause of possible harmful effects to human health or environment (e.g. endocrine disruptors) [EC, 2007b; ECHA, n.r.d] ECHA published a list of the substances that fulfil those criteria. [EC, 2007b] Authorisation procedure: The procedure of authorisation includes two steps: Step 1: Selection of substances for authorisation 54

The decision is made on the inclusion of a substance in the candidate list and further in the authorisation list (Annex XIV): o

The substance is identified as a SVHC  inclusion in the Candidate List (by Member States or ECHA; important is the public consultation process)

o

Prioritisation of the substance  Recommendation (by ECHA; important is the public consultation)

o

Inclusion of the substance in the Authorisation List (Annex XIV) (by the EC) [EC, 2007b; Schöning, 2010]

The selection process of substances for the REACH authorisation is shown in Figure 9: Figure 9. Selection of substances for the authorisation under REACH [cf. Schöning, 2010]

Step 2: Authorisation applications and decisions o

Application for the authorisation by the industry

o

Granting of authorisation by the EC (based on the opinions of ECHA’s scientific committees: the Risk Assessment Committee and the SocioEconomic Analysis Committee) 55

o

Review of authorisations (by industry, ECHA’s scientific committees and EC) [Schöning, 2010]

Figure 10. Authorisation applications and decisions [cf. Schöning, 2010]

After this 2-step process, SVHCs may become substances to authorisation by listing in the Authorisation List. A placement on market or usage after a given date of the substance is not allowed unless an authorisation. [EC, 2007b; ECHA, n.r.d]

Candidate List of Substances of Very High Concern The first step of the authorisation procedure is the identification of a substance’s possible harmful effects to human health or the environment and therefore the identification as a SVHC. A Member state or ECHA (on request of the EC) is able to propose a substance as a SVHC. Commendations and/or additions of further information of a proposed substance are allowed by anyone, whereas the submitting member state or ECHA must respond to any comments and to transfer them to the Member State Committee. The Member State Committee agrees on the SVHC identification. If the substance gets indentified as a SVHC, it will be added to the Candidate List. This list contains the candidate substances for a

56

possible entry in the Authorisation List (Annex XIV). For manufacturers, importers or users of this substance this leads to legal obligations. [ECHA, n.r.d; ECHA, n.r.e] Currently, 84 substances are on the Candidate List of SVHCs and 14 substances on the Authorisation list (updated 18 June 2012). [CIRS, 2012] The placement on the market or the use of a substance that is included in Annex XIV is not allowed for industry, unless the industry has an EC granted authorisation for this substance. [Schöning, 2010]

Restriction Through the restriction of a substance (on its own, in a preparation or in an article) which is an unacceptable risk for human health and the environment, the manufacturing, placing on market or usage are banned or restricted. Thus, restriction means any prohibition of the manufacture, usage or placement on the market. [Schöning, 2010; ECHA, n.r.f] In 2008, the Annex XV Restriction Report on BPA submitted by the United Kingdom was published. Aim of the report was the development of strategies for risk reduction for exposure situations identified in the EU Risk Assessment Report. [United Kingdom, 2008]

5.2.3 CLP Regulation On 20 January 2009 the Regulation (EC) No. 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures (also called CLP Regulation) entered into force and replaced the Directive 67/548/EEC (Dangerous Substances Directive), the Directive 1999/45/EC (Dangerous Preparations Directive) and REACH Title XI (Classification and Labelling) [Schöning, 2010] to the Globally Harmonised System (GHS) of Classification and Labelling of Chemicals. The GHS is a system of the United Nations. [EC, 2012c] Main aims of the CLP regulation are the harmonization and notification of the classification and labeling of chemical substances and the request for the use of alternative chemical names for chemical substances in mixtures. [Schöning, 2010] Classification and Labelling of BPA according to the CLP regulation BPA is classified and labelled according to the CLP Regulation as (see Figure 11): H317: May cause an allergic skin reaction (Skin Sens. 1) H318: Causes serious eye damage (Eye Dam. 1) 57

H335: May cause respiratory irritation (STOT SE 3) H361f: Suspected to damage fertility (Repr. 2) Signal Word: “Danger” Pictograms: “Exclamation mark”, “Corrosion”, “Health hazard” [ECHA, n.r.g]

Figure 11. Bisphenol A: Summary Of Classification and Labelling [ECHA, n.r.g]

5.2.4 Community Rolling Action Plan (CoRAP) The CoRAP contains the list of prioritised substances for the evaluation under REACH. Those prioritised substances are substances registered under the REACH regulation. [ECHA, 2011] The prioritisation of the substances has to be risk based: there has to be a suspicion that the production and/or usage of those substances may pose environmental risks or risk to human health. [ECHA, 2012f] The selection criteria to prioritise substances for their evaluation are regulated in Article 44(1) of the REACH regulation. [ECHA, 2011] The CoRAP gets arranged for three years and contains the substances that have to be evaluated by ECHA and the Member States and is updated once a year. Currently, the list contains 90 substances. [REACH Hamburg, 2012] The preparation of the CoRAP is done in cooperation with the Member States.

58

The substances that have to be evaluated are shared in the years 2012-2014 between the volunteering Member States: o

2012: 36 substances (by 17 Member States)

o

2013: 23 substances

o

2014: 31 substances

In February 2013 the list will become updated and changes may be introduced, whereas new substances may be added for the years 2013 and 2014. The appointed Member States have to evaluate the substance(s) within 12 months after the publication date (29 February 2012) of the final CoRAP and will prepare a draft decision for further information request for clarifying suspected risks if necessary. BPA is one of the 90 substances listed in the CoRAP 2012. It will be evaluated in 2012 by the Federal Institute of Occupational Safety and Health Division 5 in Germany, because it is suspected to be an endocrine disruptor, is used and exposed in wide dispersion, is produced in high aggregated tonnages and is used by consumers (Figure 12). [ECHA, 2012f]

Figure 12. BPA listed in the CoRAP [cf. ECHA, 2012f]

5.3 Endocrine disruptors legislation in the EU 5.3.1 Endocrine disrupting chemicals under REACH Definition of endocrine disrupting chemicals Currently, in the REACH regulation no definition of endocrine disrupting chemicals (EDC) exists. Additionally, in REACH EDCs are not addressed in CLP. [Schöning, 2010] The International Program of Chemical Safety (IPCS) of the WHO defines endocrine disruptors as “exogenous substances that alter function(s) of the endocrine system and consequently

59

cause adverse health effects in an intact organism, or its progeny, or (sub)populations”. [EC, n.r.] EDCs are able to interact with the endocrine system in the organism, at least in different possible ways: -

by imitating natural hormones like oestrogen or testosterone and thus setting similar reactions in the organism

-

by blocking of hormone receptors in body cells and thus preventing the action of normal hormones

-

by affecting hormone synthesis, transport, metabolism and excretion

Exogenous substances that are able to cause endocrine disruption are classified in two groups: -

Natural hormones: oestrogen and androgens (responsible for sexual development, naturally found in animals and humans) or phytoestrogens (naturally found in plants, e.g. in soya beans) for example

-

Man-made

substances:

industry

chemicals,

plastic

additives,

pesticides,

contraceptives or substances in hormone-replacement treatments for example.

Different groups of chemicals have already been identified as EDCs: -

Polychlorinated bisphenyls (PCBs)

-

Dioxin

-

Benzo(a)pyrene

-

Phthalates

-

Bisphenol A

-

Various pesticides (e.g. DDT, dieldrin, atrazine, tributyl tin)

-

Alkylphenols (e.g. nonylphenol)

-

Heavy metals (e.g. lead, mercury, cadmium) [EC, n.r.]

60

EDCs under REACH According to REACH, all substances including EDCs have to undergo the REACH registration when they are produced or imported in amounts of > 1 tonnes/year, but there are no specific tests for endocrine disrupting properties defined in the regular test program. EDCs will be subject to the REACH authorisation process if they are included in Annex XIV of the REACH regulation, which lists SVHCs. A clear guidance is given for CMR and PBT substances, but not for EDCs (identification only on case-by-case basis). In the case of evidence, a substance with ED properties is considered as SVHC. The inclusion or noninclusion of an EDC in Annex XIV is based on the available information and the use of a weight-of-evidence-approach5. But no internationally agreed methods or criteria are available for ED properties. [Petersen, n.r.] However, at current time there is a move in Europe to develop new strategies concerning EDCs.

5.3.2 European Commission strategy for endocrine disruptors In 1996, the EC established a policy for future research and monitoring activities in the field of EDCs. In 1999, it adopted its strategy for EDCs (“Community strategy for endocrine disruptors” - COM(1999(706)) in line with the precautionary principle to set out short-term, medium-term and long-term actions: -

Short-time actions: These actions include the collection of scientific data and substance identification for further evaluation, the identification of vulnerable population groups, monitoring levels of suspect chemicals, the establishment of a network on international level, communication with the public, as well as the establishment of a priority list

-

Medium-term actions: they have to ensure the fast and accurate testing of suspected substances (test development process directed by the OECD)

-

Long-term actions: review, update, amendment or adaption of legislation and policy for protecting human health and environment (e.g. implementation in REACH) [Petersen, n.r.; EC, 2012d]

5

Weight-of-evidence is defined as “the process of considering the strengths and weaknesses of various pieces of information in reaching and supporting a conclusion concerning a property of the substance.” [ECHA, 2010, p. 2] 61

Priority list The priority list - developed within the Strategy for Endocrine Disruptors as one of the short-time actions - contains chemicals for further evaluation of information about their ED properties. The priority list was established in four steps [EC, 1999; EC, 2012e]: 1. Compilation of a working list of suspected EDCs, containing data on their effects in humans, other vertebrates and invertebrates, and behaviour in the environment 2. Review of the available information for identification of those substances that might be highly persistent in the environment or are manufactured at high volumes or have potentially greater risks to any harmful effects. 3. Review of the substances that were identified in step 2 as persistent or are produced in high volume for the determination of the strength of their ED evidence and assignation to one of three categories: - Category 1: evidence of ED activity in at least one species in vivo - Category 2: evidence of biological activity related to ED in vitro - Category 3: no evidence of ED activity or no data available 4. Review of the Category 1 substances to decide if the possibility of the actual exposure of humans or wildlife is given. Assignation to: - high exposure concern (humans or wildlife exposure were expected) or - medium exposure concern (human exposure was not expected but wildlife) or - lowest exposure concern (human or wildlife were not exposed) [EC, 2012e]

564 chemicals had been suggested to be suspected to be ED, whereas 147 chemicals were considered to be produced in high volumes or to be persistent in the environment. A clear evidence of ED activity (Category 1) was noted for 66 chemicals in a first assessment while another 53 chemicals showed some evidence (Category 2). Humans are exposed to 60 of the 66 chemicals assigned to category 1, which includes substances like DDT and some other pesticides, polychlorinated biphenyls (PCBs), organo-tins, dioxins, styrene and phthalates. 11 substances of the working list containing 564 substances were excluded due to a lack of scientific basis for inclusion. Therefore, the candidate list contains 553 substances. [Groshart and Okkerman, 2000; EC, 2012e] Figure 13 shows the schematic overview:

62

Figure 13. Schematic overview of the project steps and the results [Groshart and Okkerman, 2000, p.3]

In 1999, the EC assigned BKH Consulting Engineers (Netherlands) to conduct a study on EDCs and to prepare a candidate list of substances based on available information for priority setting. This so-called BKH-report contains 15 annexes and was published in 2000. BPA is listed in Annex 1 (“Candidate list of 553 substances”) (No. 24, Group I). In Annex 7, the data of the ED effects of BPA on human health was evaluated at an expert meeting and listed in Annex 7 (“Human health and wildlife relevant data on endocrine disruption included in the database on 146 substances evaluated in the Expert meeting”): Figure 14 shows these ED effects of BPA relevant for human health, and Figure 15 the effects relevant for wildlife, respectively. Additionally, BPA is listed in Annex 15 (“List of 66 substances with classification of high, medium or low exposure concern”) of the BKH report as substance of high concern. [Groshart and Okkerman, 2000] 63

Figure 14. Annex 7a: Human Health endocrine disrupting effects data evaluated at the expert meeting [Groshart and Okkerman, 2000, Annex 7, p.7.22-23]

64

Figure 15. Annex 7b: Wildlife relevant endocrine disrupting effects data evaluated at the expert meeting [Groshart and Okkerman, 2000, Annex 7, p. 7.43]

65

5.3.3 Implications and current activities Implications for the regulation of EDCs It has been recognised that there is a need for a better integration of epidemiological evidence in hazard risk characterisation of chemicals. Standardised validated methods and Good Laboratory Practices (GLP) put great emphasis for the assessing the quality of toxicological studies, but they are no guarantee for quality if relevant endpoints or exposure at critical phases of development have not been included in these studies. Furthermore, at the present no universally accepted classification scheme exists. Thus, the development of a similar scheme of evidence classification and categorisation of EDCs is necessary. [Kortenkamp et al., 2011] Current activities In June 2012, the EC organised a Conference on Endocrine Disruptors to discuss the effects of EDCs on environment and human health, as well as the risks, the identification of those substances and policy objectives, because of the growing concerns. As an outcome of the conference, the EC expects to propose criteria for EDCs identification by the end of 2013, and will review the authorisation of EDCs under REACH by June 2013. Furthermore, the EC will revise their Community Strategy on Endocrine Disruptors. [EC, 2012g]

5.4 Legislation of BPA in the European Union The legislation of BPA in the EU was and is ruled in various directives and regulations. Currently, the usage of BPA in plastic and food contact materials in the EU is regulated and permitted under the Regulation (EU) No 10/2011. Since 2011, the use of BPA in plastic infant feeding bottles is prohibited, which is regulated in the Directive2011/8/EU. [EFSA, 2012a] In the EU, the use of BPA in cosmetics is banned since 2005. [Austrian Federal Environment Agency, 2010]

5.4.1 Consumer products Directive 67/548/EEC of June 1967: Directive 67/548/EEC regulates the classification, packaging and labelling of dangerous substances. Annex I of this Directive contained a list of dangerous substances. This list is 66

updated through Adaptations to Technical Processes (ATP). [EU, 2011; REACH Compliance, n.r.] In Annex I BPA was classified and labelled as (Annex I Index Number: 604-030-00-0) [IHCP, n.r.]: Repr. Cat. 3; R62 - Xi; R37-41 - R43 - R52 Reproduction toxicity Category 3; Possible Risk of impaired fertility (R62) Risk Phrases: Irritating to respiratory system (R37), Risk of serious damage to eyes (R41), May cause sensitization by skin contact (R43), Harmful to aquatic organisms (R52) Symbol and Indication of Danger: Xn (Harmful)

[IHCP, n.r.]

In 2009 this Annex I was replaced in 2009 by the Regulation (EC) No. 1272/2008 (CLP Regulation) and REACH Title XI [Schöning, 2010; EU, 2011].

Directive 2002/72/EC of 6 August 2002: The Directive 2002/72/EC regulated plastic materials and articles, which come into contact with food as part of the Regulation (EC) No 1935/2004 and lead to the cancelation of the Directive 90/128/EC. Additionally, it contained limits for the migration of substances into food. The specific migration limit (SML) of BPA was set at 3 mg/kg food. [EC, 2002]

Regulation (EC) No 1935/2004 of 27 October 2004: The Regulation (EC) No 1935/2004 contains rules about materials and articles which come into contact with food and repeals the Directive 80/590/EEC. [EC, 2004a]

Commission Directive 2005/80/EC of 21 November 2005: This Directive is an amendment to the Council Directive 76/768/EEC. BPA was added to Annex II. Therefore, BPA is prohibited in cosmetic products. [EC, 2005]

Regulation (EC) No 1223/2009 of 30 November 2009: BPA is included in the list of prohibited substances in cosmetic products (Annex II) of the Regulation (EC) No 1223/2009 on cosmetic products. This Regulation harmonised the rules in the EU. [EC, 2009a]

67

Commission Directive 2009/161/EU of 17 December 2009: The health-based and non-binding values that are derived from available scientific data the so-called Indicative Occupational Exposure Limit Values (IOELV) - set out threshold exposure levels. Below these levels no effects are expected for a substance after shortterm or daily exposure over the life time of a worker. Those IOELVs are listed in three lists: the Directives 2000/39/EC and 2006/15/EC under Directive 98/24/EC contain the fist and the second list, while the third list is established in Directive 2009/161/EU. In this third list, the limit value of BPA (inhalable dust) is set out to 10 mg/m3 of air at 20°C and 101.3 kPa for 8 hours (measured/calculated in relation to the reference period of 8 h time-weighted average). [EC, 2009b]

Regulation (EU) No 10/2011 of 14 January 2011: The Regulation (EU) No 11/2011 based on the Regulation (EC) No 1935/2004 contains rules for plastic materials and articles that come into contact with foodstuff and their safe use. This regulation repealed the EC Directive 2002/72/EC for reasons of clarity, because it was amended for 6 times. Additionally, obsolete and redundant parts were removed. The Regulation entered into force on 1st of May 2011. BPA is not authorised for the use as additive or polymer production aid and is used as starting substance or macromolecule obtained from microbial fermentation. The new SML of BPA is 0.6 mg/kg food. [EC, 2011a]

Directive 2011/8/EU of 28 January 2011: The use of BPA in plastic articles and materials that come into contact with foodstuff was allowed and regulated in the Commission Directive 2002/72/EC of 6th of August 2002. In March 2010 the Danish Government banned the use of BPA for the production of plastic materials that come in contact with children at the age of 0-3, because of the results of the risk assessment provided by the National Food Institute at the Technical University Denmark (DTU Food). In July 2010 the French Government temporarily banned the production, the import, the export and the placing on the market of BPA containing feeding bottles, because of the opinion of the AFSSA in January 2010 and the opinion of the Institut National de la Santé et de la Recherche Médicale (INSERM) in June 2010. The EC requested EFSA to evaluate the study of the Danish risk assessment and other new scientific data and concluded that no new study was identified to reverse the current TDI of BPA of 0.05 mg/kg bw/d (the detailed opinion of EFSA is discussed in Chapter 5.5.1). But because of the availability of adequate alternatives to PC infant feeding bottles containing 68

BPA, such as glass bottles, the possible particular vulnerability of the infants and the precautionary principle (Article 7 of EC regulation No 178/2002), the EC temporarily banned the use of BPA in PC infant feeding bottles. [EC, 2011b]

Regulation (EU) No 321/2011 of 1 April 2011: The Regulation (EU) No 321/2011 did not contain the restrictions of BPA in PC infant feeding bottles (Directive 2011/8/EU). Because of this, the EC adopted this Regulation: Table 1 of Annex I of Regulation (EU) No 10/2011 has to contain the information, that BPA is restricted for the use and production of PC infant feeding bottles. [EC, 2011c]

5.4.2 Environment On 16th of December 2008, the EU released the Directive 2008/105/EC on environmental quality standards in the field of water policy, in which quality specifications for priority substances and other priority hazardous substances are provided in Annex I. For BPA, no limit value exists. [Austrian Federal Environment Agency, 2010] However, BPA was named in Annex III (“Substances Subject to Review for Possible Identification as Priority Substances or Priority Hazardous Substances”) [EC, 2008, p.14] and had to be reviewed until January 2011 for its possible adaption in Annex X (“Priority Substances”) in Directive 2000/60/EC6. [Austrian Federal Environment Agency, 2010] After the review, the noninclusion of BPA in Annex X was decided. Thus, BPA is not listed as a priority substance. [EC, 2012h]

5.4.3 Summary of BPA legislation in the EU BPA is included in Part A of Annex II of the Directive 2002/72/EC in the list of authorised monomers and other starting substances (reference number: 39680), as well as in Part A of Annex III in the list of additives fully harmonised at Community level (reference number: 39680). Thus, in the EU the use of BPA is authorised for the manufacture of plastic materials and articles that come into food contact. Because of the food contact, small

6

“Directive 2000/60/EC of the European Parliament and of the Council establishing a framework for the Community action in the field of water policy”, also called EU Water Framework Directive (WFD), adopted on 23 October 2000 [EC, 2012a]. The Member States obligate themselves to achieve a good chemical condition of surface waters until 2015, in which hazardous substances do not occur in concentrations higher than regulated in the Directives. [Veltwisch, 2009] Article 16(4) provides the continuous review of Annex X, which contains the list of priority substances. Substances chosen from the list are hazardous to the aquatic environment. [EC, 2012b] 69

amounts of BPA are able to migrate from the packaging into food. For this case, a specific migration limit of 0.6 mg/kg bw/d is set out in the Directive 2002/72/EC. [AESAN, 2011] Since 2005, the use of BPA in cosmetic products is banned. [EC, 2005] On 28 January 2011 the Directive 2011/8/EU was released as amendment of Directive 2002/72/EC on restriction of BPA in infant feeding bottles. Thus, the usage of PC baby bottles containing BPA is prohibited. [AESAN, 2011]

5.4.4 Trends in Europe Denmark: In March 2010, the Danish Government passed an interim ban of BPA in food contact materials for children at the age of 0-3 years which came into force at July 1, 2010. This decision was based on a new assessment of the DTU Food and the principle of precaution. [Danish Ministry of Food, Agriculture and Fisheries, 2010; EC, 2011b] France: In March 2010, the French Government preliminary banned production, import, export and market placement of BPA in infant feeding bottles based on the opinions of the AFFSA and the INSERIM both published in 2010. [EC, 2011b] In 2011, the French National Assembly extended the ban from infant feeding bottles to all food contact materials by 2014. Additionally, they planned the ban of all packaging materials that contain BPA and aimed at children from age 0-3, as well as the inclusion of warning labels on any packaging material that contains BPA for children under the age of 3 years and pregnant women. [ITRI, 2011; USDA, 2012] As a reaction on the French ban of BPA to all food contact materials in 2014, six EU member states (Italy, Czech Republic, the Netherlands, Slovenia, the United Kingdom and Spain) objected to this plan in 2012, because it would breach the free trade rules of the EU. [European Environment & Packaging Law, 2012] Norway: In Norway BPA is regulated for its use in cosmetics and a migration limit is set out for food contact materials, but no product regulation for BPA exists. The Norwegian Government plans the prohibition of BPA in consumer products on more than 0.0025 percent by weight. [Norwegian Pollution Control Authority, 2007] Sweden: The Swedish Government will prohibit the BPA use in protective coatings in food packaging materials for children at the age of 0-3 following a report by the Swedish Chemicals Agency (KEMI) and the Swedish Food Agency. This ban will affect mainly the lids of baby feeding bottles. [RSC, 2012] In 2012, KEMI has to investigate a ban of BPA usage in thermal paper. [Swedish Ministry of Environment, 2012] 70

5.5 BPA under REACH Registration BPA is a phase-in substance and is used for the production of PC and epoxy resins. This BPA-PCs and BPA-epoxy resins are polymers, which have not to be registered under REACH. However, the BPA monomers that are the components for the polymers have to be registered.7 Deadline of the registration was in December 2010. [PlasticsEurope, n.r.a] Monomers are defined as intermediates. Thus, the registration of BPA has to follow reduced requirements. [CIRS, 2008-2012] Articles8 that are made of BPA-materials like PC or epoxy resin products (e.g. compact discs, safety helmets, automotive parts or coatings) only have to be registered by the manufacturer or importer if BPA releases from the article during a normal or reasonable use and if some other additional conditions are fulfilled. [PlasticsEurope, 2011a] Authorisation Intermediate BPA as monomer for the production of PC and epoxy resins (more than 99%) and is an intermediate.

Therefore,

BPA

is

exempted

from

authorisation

under

REACH.

[PlasticsEurope, 2010] CoRAP In the CoRAP, prioritised substances that are registered under the REACH regulation are listed for the REACH evaluation process. BPA is one of the 90 listed substances in the CoRAP 2012 because it is suspected to be an EDC. BPA has to be evaluated in 2012 by the Member State Germany. [ECHA, 2011; ECHA, 2012f]

7

The registration dossier of BPA is available at: http://apps.echa.europa.eu/registered/data/dossiers/DISS9dbe071c-c12d-0fe1-e044-00144f67d249/DISS-9dbe071c-c12d-0fe1-e044-00144f67d249_DISS-9dbe071cc12d-0fe1-e044-00144f67d249.html 8

In the REACH Directive an article is defined as “an object which during production is given a special shape, surface or design which determines its function to a greater degree than does its chemical composition” [EU, 2006, p. 57] 71

Priority list Within the Strategy for Endocrine Disruptors, a priority list was developed for further evaluation, which contains substances that have/are suspected to have ED properties. [EC, 1999; EC, 2012e] A candidate list (“Candidate List of 553 substances”) exists for further possible inclusion in the priority list. BPA is listed in this candidate list in Annex 1 (No. 24, Group I) and in Annex 7 which contains relevant data on ED of substances. [Groshart and Okkerman, 2000]

5.5.1 Scientific opinion of the EFSA One of the tasks of the European Food Safety Authority (EFSA) is the adoption of scientific opinions and independent advice for risk managers concerning to the BPA safety in food contact materials. The EFSA Panel on food contact materials, enzymes, flavourings and processing aids (CEF) carries out this work. During the past years, EFSA and their panels considered a multitude of scientific publications as well as study reports submitted from the industry. [EFSA, 2012b] In 2006, EFSA published their risk assessment of BPA and set the TDI of 0.05 mg/kg bw/d. Furthermore, they evaluated the BPA intake via food and beverages in adults, children and infants and concluded that the values were clear below the TDI. The effects of BPA in low doses were not demonstrated in a way for the use of risk assessment preparation. [EFSA, 2006; EFSA, 2012b] In 2008, EFSA adopted their opinion on the toxicokinetics of BPA in infants and adults and their ability to eliminate BPA out of the body. They confirmed that the BPA exposure to adults and infants were below the current TDI and that the metabolism and elimination of BPA in humans occur in a rapid way. The age-dependent differences in BPA toxicokinetics in humans and animals had no impacts on the EFSA risk assessment from 2006. [EFSA, 2008; EFSA, 2012b] The same year, the EC requested EFSA for the assessment of the study outcomes by Lang et al. 2008. Lang found associations between urinary BPA levels and increasing occurrences of diabetes and heart diseases. EFSA indicated the missing data on long term exposition which would be necessary to show correlations between BPA exposure and the development of heart diseases and diabetes, which are chronic diseases. For EFSA, this study did not provide evidence for these correlations. [EFSA, 2012b] On behalf of the EC, in 2010 EFSA published their opinion on a study by Stump, 2009 about the dietary developmental neurotoxicity in rats as well as on recent scientific 72

literature published in 2007 - 2010 for their relevance for the BPA risk assessment. These studies had investigated the impact of the current TDI of 0.05 mg BPA/kg bw/d. The study performed by Stump was inconclusive, had limited value, and is not able to be used for risk assessments because of the large variability in data. In conclusion, the EFSA panel did not find a new study calling for a revision of the current TDI based on their evaluation, and added that in case of a future availability of new and relevant data, they will make a reconsideration of their opinion. [EFSA, 2010] One member of the panel expressed her minor opinion which can be found in the annex of the EFSA publication. The panel member considered that some of the current studies suggested uncertainties relating to harmful health effects in doses below those used for the TDI determination. However, the panel member supported the panel opinion that these studies were not sufficient to establish a lower TDI. [EFSA, 2012b] In 2011, the EFSA CEF Panel published a statement on the reports on BPA of the French Health and Safety Agency (ANSES) and concluded that the information published in those reports on the health effects of BPA gave no reasons to change the EFSA opinion on BPA. [EFSA, 2011; EFSA, 2012b] Currently, EFSA has started with their work on a new BPA risk assessment with focus on the exposition of vulnerable population groups which is planned to be finished in May 2013. Aim is an amendment to further scientific advice by evaluating the new scientific data since their opinion on BPA published in 2006 and the uncertainties about possible negative effects of low BPA doses on human health. Furthermore, EFSA is convening an international colloquium of experts to discuss the evidence of low dose effects. [EFSA, 2012c]

5.6 International Legislation of BPA 5.6.1 USA In the USA, BPA is banned in some consumer products in several states. Since 2010, BPA is prohibited in sippy cups and baby bottles in Minnesota, Massachusetts and Wisconsin, and since 2011 in California. The first US-American city who banned BPA from cups and baby bottles was Chicago, Illinois in 2009. In Maryland, the ban of BPA in baby bottles and cups will go into effect in 2014. Vermont banned BPA from canned and bottled infant formula, plastic baby containers, reusable food and beverage containers in 2010 and New York from children products in 2010. In Delaware, BPA is prohibited in children bottles, cups and other containers of food and beverages since 2011 and since 2012 it was 73

banned from baby bottles, sippy cups, water bottles and reusable food storage containers in 2012. [Safer States, 2012]

5.6.2 Canada Canada was the first country worldwide that banned BPA from its use in baby bottles and the first country that classified BPA as hazardous to human health. Canada identified BPA as a substance with high-priority for the assessment of human health risk in 2006. [Hengstler et al., 2011] In October 2008, Environment Canada and Health Canada published their final screening assessment report on BPA. They concluded that BPA may enter into the environment in concentrations or under conditions that (may) lead to human health effects in Canada and (may) have immediate or long-term harmful effects on the environment itself or on the biological diversity. Basis for this conclusions were results of neurodevelopmental and behavioural toxicity studies in rodents. Because of this, BPA meets the criteria in paragraphs 64(a) and 64(c) of the CEPA 19999 and the criteria for persistence, but not the criteria for bioaccumulation, which are defined in the socalled “Persistence and Bioaccumulation Regulations” in CEPA 1999. [Environment Canada and Health Canada, 2008a] Health Canada concluded that the margin of BPA for the safety was too small for formula-feed infants although the results of scientific studies showed that the exposure of BPA to newborns and infants are below the levels that can cause health effects. [Morrissey, 2008] They pre-established a provisional tolerable daily intake (TDI) of 25 µg/kg bw/d as a safe level for BPA in food. In March 2010, Canada banned BPA from baby bottles and prohibited the advertisement, sale and import of these products. [Hengstler et al., 2011] In September 2010, the Canadian Government listed BPA as a toxic substance (List of Toxic Substances in Schedule 1 of CEPA 1999) [Canada Gazette, 2010].

5.6.3 Australia and New Zealand Food Standards Australia New Zealand (FSANZ)10 has evaluated the safety of BPA in baby bottles. They concluded that the levels of exposure of BPA are low and there is no

9

Canadian Environmental Protection Act, 1999. It came into force on 31 March 2000. Aims are the pollution prevention, the protection of human health and environment. [Environment Canada, 2012] 10

Established in 1991, FSANZ is a bi-national independent government agency as part of the Government’s Health and Ageing portfolio of Australia. Task of the FSANZ is the development of effective food standards in collaboration with the governments in Australia and New Zealand. [FSANZ, 2012] 74

risk for infant health. But because of consumer concerns and as response to approaches taken by governments in other countries (not because of product safety), they introduced a voluntary phase-out of PC baby bottles containing BPA by major retailers in July 2010. [FSANZ, n.r.; Hengstler et al., 2011]

5.7 Opinions by Stakeholders and Authorities 5.7.1 U.S. FDA The U.S Food and Drug Administration (FDA) is the consumer protection agency of the U.S. Department of Health and Human Services. Tasks are the protection of the public health, the controlling of the safety and efficacy of human and veterinary drugs, biological products, food, medical devices, cosmetics and radiation emitting products. [FDA, 2010] The FDA released its document “Draft Assessment of Bisphenol A for Use in Food Contact Applications” in August 2008, which was reviewed by a Subcommittee of FDA’s Science Board and published its report in October 2008. Additional low-dose toxicity studies were reviewed by the Center for Food Safety and Applied Nutrition within the FDA. As result of these reviews, they released the document “Bisphenol A: Review of Low Dose Studies” in August 2009. [FDA, 2012] FDA shares the perspective of the NTP (cf. Chapter 5.7.2). Studies provide some concern of possible effects of exposure of BPA to behaviour, brain and prostate gland of fetuses, infants and children. However, the FDA also “recognizes substantial uncertainties with respect to the overall interpretation of these studies and their potential implications for human health effects of BPA exposure. These uncertainties relate to issues such as the routes of exposure employed, the lack of consistency among some of the measured endpoints or results between studies, the relevance of some animal models to human health, differences in the metabolism (and detoxification) of and responses to BPA both at different ages and in different species, and limited or absent dose response information for some studies.” [FDA, 2012, p. 3] Because of those uncertainties, FDA runs additional studies and supports the reduction of BPA exposure to humans. But FDA makes no recommendations to change food or infant formula use. The support of the reduction of BPA exposure of infants in the food supply and the support and evaluation of production practices of BPA and alternative substances are precaution steps, but the FDA “is not recommending that families change the use of infant formula or foods, as the benefit of a stable source of good nutrition outweighs the potential risk of BPA exposure.” [FDA, 2012, p. 6] 75

The FDA supports actions of the industry to stop the manufacturing of bottles and infant feeding cups containing BPA for U.S. market and the development of BPA alternatives for infant formula cans. It will review new studies as well as other relevant material and will update its BPA assessment when significant new information is available. [FDA, 2012]

5.7.2 NTP The NTP (National Toxicology Program) is an inter-agency program that was established in 1978 by the Secretary of Health, Education and Welfare, which is today known as the Department of Health and Human Services and evaluates agents of concern to public health. The core of the NTP consists of three agencies: the National Institute of Environmental Health Sciences (NIEHS) of the National Institutes of Health (NIH), the National Institute for Occupational Safety and Health (NIOSH) of the Centers for Disease Control and Prevention (CDC) and the National Center for Toxicological Research (NCTR) of the FDA. [NTP, 2005] In 2011, the NTP Office of Health Assessment and Translation (OHAT) was established. It serves as a resource of environmental health to various agencies and to the public, and it accomplishes evaluations for the assessment of the evidence of harmful effects of environmental substances, which are published as NTP Monographs. It also provides opinions. Formerly, those assessments were carried out by the NTP Center for the Evaluation of Risks to Human Reproduction (NTP-CERHR) from 1998 to 2010. [NTP, 2011] In September 2008, the NTP published its final report the on BPA (“NTP-CERHR Monograph on the Potential Human Reproductive and Developmental Effects of Bisphenol A”), that provides the current NTP opinion on BPA and its possible harmful effects on human’s reproduction and development. [NIEHS, 2008] The report contains a NTP brief of BPA and an Expert Panel Report on the reproductive and developmental toxicity of BPA. The NTP concluded that there are some concerns about current human exposure of BPA for effects on brain, behaviour, and prostate gland in fetuses, infants and children, minimal concern about effects of BPA on mammary gland and earlier age for puberty for females, and minimal concern about workers who are exposed to higher BPA levels. The NTP has negligible concern about fetal or neonatal mortality, birth defects or reduced birth weight as well as growth in the offspring of pregnant women, who are exposed to BPA, and that a BPA exposure cause effects on reproduction in non-occupationally exposed adults. The NTP added that these conclusions are based on the available information when the report was prepared. [NTP-CERHR, 2008]

76

5.7.3 U.S. EPA The Environmental Protection Agency of the United States (U.S. EPA) was established in 1970. Its aim is the protection of human health and environment. [U.S. EPA, 2012a] In its “Bisphenol A Action Plan”, U.S. EPA reviewed BPA and considered potential actions. Because of the effects of BPA on reproduction and development shown in animal studies, its weak estrogenic outcomes and the uncertainties on low-dose effects, questions about the potential impact of BPA on human health and environment exist. Thus, the U.S. EPA intends to consider rulemaking under the Toxic Substances Control Act (TSCA)11. Additionally U.S. EPA plans the initiation of assessment activities on collaborative alternatives. But this time, based on the current knowledge of the risks of BPA to human health, EPA does not intend to initiate regulatory action under TSCA and continues to consult and coordinate with the FDA, the CDC, and the NIEHS for a better assessment and evaluation of the potential health effects of BPA. [U.S. EPA, 2010; U.S. EPA, 2011]

5.7.4 WHO and FAO A large number of animal studies on toxicity and endocrine activity of BPA has been published in recent years. Some of them were performed in compliance with OECD guidelines and many others without. Because of the discrepancies in the outcomes of those studies, a controversy about BPA and its safety for human health exists. [INFOSAN, 2009] Thus, in 2010, the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organisation (WHO) held an expert meeting to review the toxicological and health aspects of BPA with more than 30 experts supported by the EFSA, Health Canada, NIEHS and the FDA. [WHO, 2012] The WHO/FAO Expert Meeting concluded that for many endpoints estimated in various studies there is no health concern because the points of departure are higher than the BPA exposure to humans. Effects on development and reproduction were only shown at high doses of BPA. However, in a few studies correlations between lower BPA levels and neurodevelopment, changes in mammary glands and prostate in rats and sperm parameters were shown. Furthermore, the WHO/FAO Expert Meeting concluded that “it is difficult to interpret these findings, taking into account all available kinetic data and current understanding of classical estrogenic activity. However, new studies indicate that BPA may 11

The TSCA (passed in 1976) is the main chemical law in the USA to regulate the use of chemicals. [Safer Chemicals, 2009] It authorizes the EPA to require reporting and testing requirements, as well as restrictions of substances. [U.S. EPA, 2012b] 77

also act through other mechanisms. There is considerable uncertainty regarding the validity and relevance of these observations. While it would be premature to conclude that these evaluations provide a realistic estimate of the human health risk, given the uncertainties, these findings should drive the direction of future research with the objective of reducing this uncertainty.” [FAO and WHO, 2010, p. 30]

5.7.5 Environment Canada and Health Canada Environment Canada and Health Canada are departments of the Government of Canada. In 2006, BPA was identified as a high-priority substance for human health risk assessment by the Canadian Government. [Hengstler et al., 2011] In 2008, Health Canada and Environment Canada published its Risk Management Approach for BPA and indicated that actions were needed because BPA is considered “as a substance that may be entering the environment in a quantity or concentration or under conditions that constitute or may constitute a danger in Canada to human life or health.” [Environment Canada and Health Canada, 2008, p. 6] They established a new provisional TDI of 25 µg/kg bw/d BPA in food. In 2010, the use of BPA in baby bottles was prohibited [Hengstler et al., 2011] and listed as a toxic substance. [Canada Gazette, 2010] Thus, Canada was the first country who classified BPA as hazardous to human health.

5.7.6 ANSES The French Agency for Food, Environmental and Occupational Health & Safety (ANSES) was founded in 2010 following the merger of the French Agency for Environmental and Occupational Health Safety (AFSSET) and the French Food Safety Agency (AFSSA), which have been two health agencies in France. [ANSES, 2011] In 2009, AFSSA published their opinion on BPA. It reported that the determination of the significance of warning signals observed on various studies on low-dose effects of BPA is necessary and that an increase of the safety factor of the current TDI should be debated. Furthermore, the investigation of BPA sources other than food contact materials should be carried out and a methodology for the assessment of potential risks to human health of very low doses should be developed. [AFSSA, 2010]

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5.7.7 German BfR The Federal Institute for Risk Assessment (BfR) is part of the German Federal Ministry of Food, Agriculture and Consumer Protection and is independent in respect to its research and assessments. The tasks of BfR are the detection and assessment of new risks for human health, working out recommendations on risk reduction and the communication. Results of its work are the basis for scientific advice to federal ministries and other agencies. [BfR, 2012] In 2008, BfR published an opinion on BPA and pointed out that the investigation of the toxicity of BPA is adequate. BPA has a low acute toxicity and there is no evidence of carcinogenic effects. Though, BPA has weak estrogenic effects and is an endocrine disruptor. However, in human body BPA is metabolised in a fast way, whereas in rodents the BPA metabolism occurs slowly and hormonal effects were shown in animal studies. The BfR supported the assessments of the EFSA. [BfR, 2008]

5.7.8 German UBA The German Federal Environment Agency (UBA) was founded in 1974. The key tasks are the scientific support to the federal Government, the implementation of environmental laws and to public information about environmental protection. [German Federal Environment Agency, 2012a] The German UBA published a background paper on BPA. Because of the existing studies on the BPA effects and exposition there is advice of possible risk for human health. Some aspects of risk assessment show gaps in knowledge and uncertainties at the present time. The German UBA considered precautionary provisions for the reduction of BPA exposure for vulnerable population groups. The available data of studies published in renowned journals should be regarded in a suitable way. The German UBA pointed out that it is important to act precautionary and to restrict the usage of particular products, especially food contact materials. Additionally, because of the available information about BPA in the environment no final assessment of the environmental risks of BPA is possible. The environmental risks may be underestimated. [German Federal Environment Agency, 2010b] The German UBA will accomplish a further assessment of BPA in 2012, in the framework of the REACH regulation. The possible harmful effects of BPA on hormone systems will be investigated and emissions into the environment will be specified. [German Federal Environment Agency, 2012b] 79

5.7.9 Non Governmental Organisations (NGO’s) Global 2000 (Friends of the Earth Austria) “Friends of the Earth International” is a global network with more than 5,000 local activist groups in 71 states worldwide with its head office in Amsterdam, Netherlands. Global 2000 with its office in Vienna, Austria is an independent environmental NGO, and since 1999 one of the members of “Friends of the Earth International”. Main issues are e.g. environmental protection, agriculture, gene technology, sustainable development and climate. Global 2000 acts through activism, lobbying and releasing information for public and politics. [Global 2000, 2010a; Global 2000, 2011] In the recent years, Global 2000 indicated the risks of BPA in baby bottles and baby soothers as well as in food contact materials, and claimed further investigations as well as a ban to BPA in food contact materials and baby articles. [Global 2000, 2010b]

BUND (Friends of the Earth Germany) Founded in 1975, BUND (Bund für Umwelt und Naturschutz Deutschland) is a German environmental NGO and part of Friends of the Earth International [BUND, n.r.]. In 2008, BUND published a report on BPA in which it was indicated that there is an evidence for ubiquitous BPA exposure that have to lead to decisions to reduce this exposure. The BfR as well as EFSA should consider new studies for their risk assessment and decisions should not be provided by the industry. Additionally, BUND requested, that BPA manufactures should not register applications for food contact materials and baby bottles under REACH, and advice that consumers should not use plastics of PC. [Wefers and Cameron, 2008]

International Chemical Secretariat (ChemSec) The International Chemical Secretariat (ChemSec) is an NGO based in Göteborg, Sweden that was founded in 2002 by the four environmental organisations Swedish Society for Nature Conversation, WWF (Word Wildlife Fund) Sweden, Nature and Youth and Friends of the Earth Sweden, and is working towards a toxic-free environment. [ChemSec, 2012a] ChemSec indicates that there is a need to regulate EDCs like BPA in the EU. The process of REACH is moving very slowly, and it is time to act on EDCs. [ChemSec, 2012b] Regarding to BPA, ChemSec advice consumers to use BPA-free baby bottles, to eliminate PC food contact materials and drinking containers, as well as the consumption of canned 80

food and drinks, to avoid oral contact with thermal paper and to avoid products made with BPA-related substances. [ChemSec, 2012c]

5.7.10 Industry In Europe, the official voice of European plastics producers is PlasticsEurope with head office in Brussels, Belgium. It is the European trade association and has partnerships representing the plastics manufacturing chain in Europe. [PlasticsEurope, n.r.b] After the ban of BPA in PC baby bottles by the EU in 2011, PlasticsEurope indicated that it “is concerned about the motivation of such decision that is not backed by any scientific evidence, and contradictory to the recent recommendation of EFSA (European Food Safety Agency). As such, PlasticsEurope strongly supports and advocates for a fact and sound science risk assessment based decision making process, in particular when it comes to product safety, in the best interest of the consumer, of the European industry and of society as a whole.” [PlasticsEurope, 2011b] On the industry’s website12 on BPA, they indicate that BPA is safe for its use basing on scientific studies [American Chemistry Council, 2003-2012b].

5.8 Controversies on BPA Many controversies to BPA exist between experts, regulatory agencies, industries, the press and others. [Vandenberg et al., 2009] In the following, some of a multitude of controversies are discussed.

5.8.1 Dose response curves Monotonic dose response curves show that high doses are expected to be the cause of a greater harm. If there are no effects at high doses of a substance, no effects will be expected at lower doses. This model is used for the risk assessment of substances. Thus, doses of substances below their determined NOAEL are considered as safe. But several

12

Organised by the American Chemistry Council, PlasticsEurope and the Japan Chemical Industry Association, and sponsored by the Polycarbonate/BPA Global Group (available at www.bisphenola.org) 81

studies reported that the dose response curves of hormones are U-shaped or inverse Ushaped, which indicate effects at low and at high exposure levels. They are called “nonmonotonic” dose response curves. They are explained by a down-regulation of hormone receptors at higher levels of the hormones, or by the combination of two or more monotonic curves through different pathways. Studies have shown that the dose response curves of BPA are non-monotonic. [Vandenberg et al., 2009] For example, Jenkins et al., 2011 investigated the role of the oral BPA exposure on mammary carcinogenesis in mice. They reported that significantly accelerated mammary tumor genesis and metastasis were shown at low doses of BPA in mice, but not at high doses. Thus, they concluded, that a chronic oral BPA exposure results in a non-monotonic dose response in mice. [Jenkins et al., 2011] On the other hand, in their scientific opinion EFSA reviewed the occurrence of nonmonotonic dose response curves of BPA and concluded that most of the studies that reported such findings have a lack of dose responses, limitations in their experimental design or other shortcomings. Thus, EFSA is not aware of clearly reproducible adverse effects expressed by low doses of BPA only. [EFSA, 2010]

5.8.2 Low-dose effects and mechanism for low-dose action Low-dose effects of EDCs are effects that are reported at doses lower than those which are used in traditional toxicological studies for risk assessments. The lowest dose for BPA in its risk assessment was 50 µg/kg/d [Vom Saal and Hughes, 2005]; low dose of BPA is defined by the NTP as concentrations ≤ 5000 µg/kg bw/d [FDA, 2008]. But in recent years many studies were published that investigated the effects of BPA at doses below the reference dose of 50 µg/kg/d. Vom Saal and Hughes, 2005 reviewed 115 published studies in 2004. 94 of those low-dose studies reported significant effects and further 31 studies reported effects caused by BPA below the reference dose. Some of the reported effects in mice and rats include an increased postnatal growth, early onset of the sexual maturation in females, increased prostate size in male offspring, the stimulation of mammary gland development, behavioural effects, changes in the brain, altered immune functions and decreased antioxidant enzymes. [Vom Saal and Hughes, 2005] The TDI of BPA set out by the EFSA in 2006 is 50 µg/kg bw/d. According to EFSA, there is no evidence to amend the value of the current TDI. [EFSA, 2010] But EFSA is convening an expert colloquium for a discussion of the evidence of low doses [EFSA, 2012c].

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Thus, there are many controversies between agencies, scientists and other stakeholders about low-dose effects of BPA and the safety of the current TDI. It is important to perform adequate studies of low-dose effects and mechanisms of BPA, that have to be considered in risk assessments and setting out safe limit values.

5.8.3 Inactivation through conjugation and relevance of animal studies for humans Studies of the metabolism of BPA have shown that in rodents, monkeys and humans BPA becomes rapidly absorbed in intestinal tract, and undergoes a fast conjugation to BPAglucuronide (BPA-G). The metabolism and toxicokinetics are described in Chapter 4.2. Further, BPA-G gets eliminated via urine. [FDA, 2008] In rodents, BPA is metabolised in a different way than in humans; it undergoes the enterohepatic circulation in rats and mice, while in humans this does not occur. [Völkel et al., 2002; Doerge and Fisher, 2010]. In rodents, the BPA-G elimination is complicated, because of its route into bile rather than into urine. BPA-G enters the intestines and can become hydrolysed to free BPA again by bacterial glucuronidases in intestinal tract. BPA is able to be reabsorbed and re-circulated (less than 50%), and additional free BPA may be available in the rodent models comparing to humans. Free BPA is estrogenically active. Thus, considerable uncertainties about the pharmacokinetics data on BPA exist, which have to be regarded in the risk assessment of BPA and the evaluation of its safety. [FDA, 2008] Other scientists claim that there are studies available, that not all of the BPA gets conjugated in liver. There are indications that human glucuronidases are produced in the digestive tract, and the production increases throughout infancy. Adult levels are reached in humans at the age of 4. Thereby, it may be possible that conjugated BPA be deconjugated and activated in infants during the digestive process. Further, it may be possible that conjugated BPA becomes locally deconjugated in other body tissues, but further studies are needed. Those indications lead to the question and controversies, if rodent models are adequate for the understanding of the BPA metabolism in humans. [Vandenberg et al., 2009]

5.8.4 Oral route of exposure Many studies suggest that there may be differences in the BPA metabolic pharmacokinetics after oral, subcutaneous, and intravenous exposures. The routes of exposure may affect the BPA circulating levels. On the other hand, several studies have 83

shown that there are less differences in BPA metabolism and excretion basing on the route of exposure, which suggest that the kinetics of BPA metabolism are similar regardless the exposure mode. Humans are exposed to BPA not only through the oral route. It may also occur through bathing in contaminated water, inhalation of contaminated air, and through leaching from medical devices and tubing. Thus, Vandenberg et al., 2009 indicated that it is unjustified to argue with the route of exposure in the evaluation of human risk of BPA exposure. [Vandenberg et al., 2009]

5.8.5 Use of scientific studies for risk assessment In publications by the industry, it is stated that the BPA exposure to humans is more than 400 times lower than the reference dose of 0.05 mg/kg bw/d set out by the U.S. EPA and that BPA does not have any risk for human health, because of those low exposure levels. [Vandenberg et al., 2009] The U.S. FDA, as well as the EFSA in Europe evaluated studies on potential effects of BPA to human health and decided that current levels of BPA exposure are considered to be safe. These decisions are based on studies that were conducted using Good Laboratory Practices (GLP)13. Though, the agencies did not consider hundreds of scientific studies in animals and cell cultures in their risk assessment performed by government and academic scientists who reported harmful effects of BPA at low doses, because they were not conducted using GLP. Many scientists criticize those decisions, because of their misguidance is resulting in a continued risk to humans from BPA exposure and other chemicals. The FDA and the EFSA published documents that demonstrate that their decision on BPA safety were basing primarily on study results followed GLP guidelines and that were industry-funded. [Myers et al., 2009] This decision stands in contrast to the conclusions of a panel of 38 experts at a conference supported by the NIH, U.S. EPA, and Commonweal who published “The Chapel Hill Consensus Statement” [vom Saal et al., 2007], as well as the conclusions of several peer-reviewed articles, that had reviewed hundreds of articles representing a comprehensive review of the scientific literature. The scientists believe that the industry sponsored studies according to GLP are not eligible to detect low-dose effects of BPA and other endocrine-disrupting chemicals. They claimed that GLP is not always a guarantee for reliability or validity of results of scientific studies.

13

GLP is a rule for the conduction of research on health effects or drug and chemicals safety testing submitted by private research. It outlines guidelines for conduction scientific research, and was developed as a response to misconduct by private research companies. The first rules for GLP were issued by the U.S. FDA in 1978 after a federal investigation of laboratory practices of private research companies. [Myers et al., 2009] 84

[Myers et al., 2009] Additionally, after the draft assessment on reproductive and developmental toxicity and carcinogenicity of BPA released from the FDA in 2008, in which the FDA upheld their current BPA safety standard, and in which only two industry-founded multigenerational GLP-studies were the base, the external FDA Science Board Subcommittee on BPA reviewed this FDA report. It disagreed with the decision of the FDA to exclude hundreds of peer-reviewed studies from their review that were non-GLP studies. The subcommittee concluded that the FDA failed. [Vogel, 2009]

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6 UM-MuKi study The aim of the UM-MuKi (environment-mother-child) study was to assess the presence of environmental pollutants, like BPA via human biomonitoring. The study was performed by the Environment Agency Austria (Umweltbundesamt) and the Medical University of Vienna, Austria as an implement within the European Territorial Co-Operation Programme AustriaSlovakia 2007-2013, and was funded by the European Regional Development Fund, and started in September 2009. 200 blood samples of pregnant women and cord blood of their newborns were collected at the Semmelweis-Klinik in Vienna and the University Hospital Bratislava, Slovakia, to assess exposure of newborns with human biomonitoring. BPA was analysed in 40 of the 200 collected mother-child-pairs.

6.1 Human biomonitoring The method of human biomonitoring (HBM) is used for the determination and control of human exposure to pollutants and for protection of human health. HBM is an important method for exposure and risk assessment. Concentrations of environmental chemicals are measured in various body tissues and fluids, like (amongst others) urine, blood, breast milk, cord blood, or organ tissues. Measurements can be performed in the general population, different population groups, or in individuals. Benefit is the ability to reveal the inner exposure or the individual body burden. At individual level, there are strong differences in adsorption, pharmacokinetics, metabolism and elimination of environmental chemicals, which are considered by HBM [Angerer et al., 2007; German Federal Environment Agency, 2010c].

6.1.1 BPA exposure of mothers and their newborns Several studies have investigated BPA concentrations in maternal blood samples and in cord blood samples of the newborns. Table 10 shows the results of various studies. Fénichel et al., 2012 found BPA in all cord blood samples (n=152). Lee et al., 2011 compared BPA levels in pregnant women and their newborns. BPA concentrations were mostly higher in women than in corresponding fetuses. They found BPA in 40% of the fetuses. Additionally they found positive associations between BPA levels in maternal blood and birth weights of their newborns. In their study, Schönfelder et al., 2002 found BPA in all fetuses (n=37). In 14 mother-child-pairs, the BPA concentrations were higher in fetuses than in the corresponding mothers. Those and other results suggest that BPA is able to be transferred from pregnant women into their fetuses. 86

Table 10. BPA concentrations in blood samples of mothers and cord blood samples of their newborns assessed in several studies Population (n)

Year of collection

Maternal BPA [ng/ml]

Cryptorchid (46) and noncryptorchid (controls, 106) male newborns from France

unknown

Not measured

Newborn BPA [ng/ml]

LOD

Method

Results

Ref.

RIA

BPA was found in all cord blood samples which suggest the transfer via placenta and fetal exposure. In control and cryptorchid groups similar BPA levels were measured.

[1]

(DR)

Cryptorchid:

0.08

0.14-5.75, range

(100%)

0.92, median 1.26±1.13, mean±SD Controls: 0.14-4.76, range 0.86, median 1.12±0.86, mean±SD

Mothernewborn pairs (97) from Taiwan

2006/07

Pregnant women (300) and their fetuses (300) from Korea

unknown

5.4±6.3, mean±SD

1.1±2.2, mean±SD

0.13

HPLC-UV

HPLC-FD

[2]

0.5, GM

2.5, GM ND-66.48, range

ND-8.86, range

0.625

2.73, median