www.ofi.at

Österreichisches Forschungsinstitutfür Chemie und Technik

Austrian Research Institute for Chemistry & Technology

CONSTRUCTION PLASTIC PRODUCTS

SURFACE TECHNOLOGYBIOENERGY

PHARMA

CERTIFICATION

www.ofi.at

Member of

Locations & Scope of Work

Location

1030 Vienna, Arsenal

Applied Polymer Technology

Surface Technology

Building & Construction

Sports technology

Location

1110 Vienna, Brehmstraße

Packaging

Pharma & Medical Devices

Bio energy

Food & Feed analysisstarting summer 2010:

New location

at TFZ Wr. Neustadt

www.ofi.at

Member of

Our Focus

Polymer TechnologyProcessing & application of polymers,

elastomers, paints, coatings and adhesives

Building & ConstructionBuilding materials, structural monitoring, dehumidification of masonry, sports technology

Pharma & PackagingTrace analysis, stability studies,

packaging development; food & feed analysis

Biomass fuelsManufacturing method, quality assurance,plant design

www.ofi.at

Member of

Agenda

What can you expect?

Current discussion

Definition – what are EDCs?

Assay methods and first results

What can we do?

www.ofi.at

Member of

Studies on BPA

NTP (NIH) USA 09/2008: At current exposure of the U.S.

population:

Developmental toxicity to fetuses / infants : „some concern for

adverse effects (brain, behavior, prostate)“

Federal Office of Public Health (Switzerland) (2009): „No risk to

consumers“

Problem of evasion to other / worse characterized ingredients

Statement of the Endocrine Society 06/2009: „EDCs are a

significant concern to public health“

Statement of the BfR 10/2009: „No health risk. No risk to infants

and young children.“

www.ofi.at

Member of

Actual situation

EDCs in contact with food – plastic, paper, laminates, coated metal cans...

Effects of EDCs in packaging :

• Potential danger (but not proven)

• Currently concentration on a few substances

• Large number of potential EDCs in packaging

Need for action:

• Analysis (Bioanalytsis; chemical analysis)

• Toxicological studies

• There are no comprehensive studies on EDCs in packaging

• Reduction of the EDC burden of packaging

www.ofi.at

Member of

Definition ED

www.ofi.at

Member of

Endocrine Disruptors (EDCs)

Definition:

EPA

Exogenous substances that act like hormones and disrupt the physiologic function of the endocrine system

European comission

Exogenous substances that act like hormones, disrupt the physiologic function of the endocrine system and cause adverse health effects.

www.ofi.at

Member of

Effects on the environment

Change of sex organs

Purple snail (TBT)

Alligators (DDT, Dicofol)

Accumulation during food chain

Example. PCB:

Polar bear: 3 billion times the concentration originally found in water

www.ofi.at

Member of

EDCs in plastic

Monomers, stabilizers, plasticizers, antioxidants,

contaminants ...

Bisphenol A

Alkylphenols (Nonylphenol)

Phthalates

Problem: migration out of the plastic into the food

www.ofi.at

Member of

Why estrogens?

www.ofi.at

Member of

Xenoestrogens

Act like the primary female sex hormone estrogen

Estrogen is involved in the regulation of many sensitive

developing steps and metabolism functions in both men and

women

A disfunction of these mechanisms can cause reproductive

problems, developmental disorders or cancer

www.ofi.at

Member of

German study: high estrogenic activity in mineral water

Authors assume that plastic

bottles are the source

BUT: other reasons possible

Highly contraversal study

Direct analysis of the plastic bottles necessary (PET-Bottle,

screw cap)

www.ofi.at

Member of

Selection of bioassays

• Yeast-Bioassays (YES / YAS)

• Aspergillus Screen (highly sensitive

reportersystem)

• Cell cultures (breast cancer-, prostate cancer

celllines) – E-Screen

• Reproductiontests with Potamopyrgus

antipodarum

• ...

www.ofi.at

Member of

Bioassays

Reporter gene assays:

• YES – Yeast Estrogen Screen (2 different

strains)

• Aspergillus nidulans - Bioassay

Principle

• Cloned gene for human estrogen receptor

• Binding of estrogen or endocrine disruptor (e.g. plastic

additive) activates receptor

• Expression of ß-Galaktosidase

• Conversion of CPRG (yellow) => CPR (red)

• Photometric measurement

www.ofi.at

Member of

Effects of estrogens

www.ofi.at

Member of

Reportergen-Assay

www.ofi.at

Member of

Extraction vs. migration

Total extraction

• ASE – Accelerated Solvent Extraction

Food simulants

• Water

• 3% Acetic acid

• 50% Ethanol

• Isooctane

www.ofi.at

Member of

17ß-Estradiol standard curve

Limit of detection: 10 pM

Limit of quantification: 20 pM

www.ofi.at

Member of

First results

Materials total negative positive (pg EEQ* / g sample)Limit of detection[pg EEQ/g sample]

PP

120

Yeast strain 1 23 22 1 400

Yeast strain 2 23 23 0

HDPE

Yeast strain 1 13 12 1 2000

Yeast strain 2 13 12 1 1000

6PET

Yeast strain 1 19 17 2 10 / 50

Yeast strain 2 19 14 5 50 - 80

*EEQ… 17ß-estradiolequivalent

www.ofi.at

Member of

Plasticadditives

Positive HDPE-sample: Bisphenol A: ca. 20 - 40 µg / g sample

Benzylbutylphthalat: ca. 200 - 400 µg / g sample

www.ofi.at

Member of

Plasticsamples

12 PET-bottles and PET-bottles preforms

4 PET-foils

13 screw caps

3 recycling flakes

6 positive samples, relatively low activities

www.ofi.at

Member of

Results

www.ofi.at

Member of

Interpretation

Estrogen active substances can be found in plastics

Low activity, but synthetical estrogens have a higher risk

potential than natural estrogens

Experiments with food simulants: no activity found

In vitro - studies give no information on actual effects in

humans

www.ofi.at

Member of

Why bioassays ?

Advantages

Measurement of known and unknown substances

Integration of synergistic effects (mixing effects)

Evaluation of the complete package

Simple screening tool

Evaluation of toxicological effects possible

No direct hints on in vivo effects

www.ofi.at

Member of

Bioassays

Disadvantages

False negative results possible

No conclusion about possible in vivo effects

• only show: hormone binding to receptor

Technical problems (solubilities / extracting agents / food simulants ...)

No information of which substances are causing the hormone activity

Combination with chemical analytic

www.ofi.at

Member of

Chemical Analysis

Chemical analysis

• Development of testing methods for endocrine

disruptors in packaging / food

• Goal: Development of a multimethode for the 50 most

frequent EDCs

• GC/MS

• LC/MSn

www.ofi.at

Member of

Next step: E-screen

In-vitro bioassay

Human breast cancer cells (MCF 7 cells)

No genetical modification

Reproduction of the cancer cell line dependents on estrogens

Reproduction of the cells (proliferation) is determined in comparison to a negative control and to estrogen standards

high sensitivity (ca. 1 pmol/l)

www.ofi.at

Member of

www.ofi.at

Member of

COIN

COIN program line „structure"

Funding authority: FFG on behalf of bmvit and BMWFJ

Dimension: national

Goals of the program• Development and improvement of key competences and functions

• for providers of application-oriented F&E&I-expertise

• particularly towards the KMU

Runtime: 01.09.2010 until 30.08.2014 (48 months)

Volume of the project: 1,8 Mio.€

www.ofi.at

Member of

Goal:

Establishment of new bioanalytical methods

Characterization of the total hormone burden of food

contact materials (plastics, coated metal packaging,

paper)

Bio-Assay-Battery

Sensitivity, standardization and high-throughput-analytic

Chemical detection for the 50 most important EDCs in the

ppb-range (preferable GC/MS)

www.ofi.at

Member of

Advisory board

International Scientific Advisory Board

• Toxicologists

• Food / Packaging analysts

• Reference laboratory

• Authorities

Project Advisory Committee

• Project partners (institutions)

• Participating companies

www.ofi.at

Member of

Partners / Participating Companies

The following companies already joined the project : Plastics Europe Deutschland Tetra Holdings GmbH NÖM MAM Babyartikel GmbH Verein für Konsumenteninformation (VKI) REWE Teich AG ALPLA SIG Combibloc

The following companies have expressed their interest: Greiner Packaging EREMA Miraplast MM Karton

www.ofi.at

Member of

For any questions please contact:

DI Dr. Johannes Bergmair(DW 976, E-Mail: johannes.bergmair@ofi.at)

- Hotline

ofi Research InstituteBrehmstraße 14A1110 Vienna+43-(0)1-798 16 01 -DW+43-(0)1-798 16 01- 480