S72 Abstracts / Toxicology Letters 238S (2015) S56–S383

of 5% HPAA and 5% polyacrylic acid had been added accidentally to activation system. In conclusion, it is necessary an exhaustive study
the drinking water supply resulting in the exposure of all employ- of this of kind compounds before their use in food industry.
ees with HPAA and PAA by drinking tab water, preparing coffee or
Acknowledgments: The Spanish Ministry of Science and Innova-
tea and/or skin exposure (washing, showering). Whereas PAA com-
tion (AGL2012-38357-C02-01) co-financed by FEDER Funds, and
monly is used in detergents, disposable diapers or recreational gels,
Junta de Andalucía (AGR-7252).
and its safety profile has been compiled by the German MAK com-
mission, no toxicity data exist for HPAA. Inhalation of PAA resulted http://dx.doi.org/10.1016/j.toxlet.2015.08.250
in mucosal irritation at higher concentrations (>0.1 mg/m3 ), no
adverse effects were observed after oral intake in experimental
animals. For HPAA no data about its toxic potential are availabe. P02-026
Toxnet (toxnet.nlm.nih.gov) did not report relevant hits, so any Fingerprinting (Py–GC/MS) of a bio-film active
estimate of the health effects of HPAA must rely on analogy. A food package with Origanum vulgare L.
structurally related compound, foscarnet (phosphonoformic acid), essential oil
is used as an antiviral compound and exerts its effects by binding to
the pyrophosphate pocket; medical usage requires 40–120 mg/kg M. Llana-Ruiz-Cabello 1 , S. Pichardo 1 , N. Jimenez Morillo 2 , J.M.
body weight. In order to assess possible health effects of HPAA all Bermudez 3 , S. Aucejo 3 , F. Gonzalez-Vila 2 , A.M. Cameán 1,∗ , J.A.
employees were offered a health check after the contamination González-Pérez 2
was determined, which was accepted by 70%. This health check
1 Area of Toxicology. University of Sevilla, Sevilla, Spain
included selected laboratory parameters and a focused patient his-
2 IRNAS-CSIC, Sevilla, Spain
tory; these data currently are evaluated. Preliminary results do not
3 Area of Packaging Materials and Systems, ITENE, Valencia, Spain
indicate overt acute toxicity; among the laboratory values no single
parameter corresponded with HPAA uptake. Appr. 10% of the lab-
oratory values showed major deviation of at least one parameter The use of biobased packaging materials are becoming an
from the reference range; these deviations currently are assessed as alternative to polymers produced from non-renewable resources.
to the causation. We will present additional results from the ongo- Moreover, increasing interest in the use of active additives from
ing analysis of both the laboratory values and the clinical history natural sources has been observed in consumers and industry.
data. Oregano essential oil (EO) is being included in new materials due
to its bioactive properties. However, due to EOs high volatility,
http://dx.doi.org/10.1016/j.toxlet.2015.08.249
aspects like ageing and conditions of films preparation may
cause losses of active agent. In this work, analytical pyrolysis
(Py–GC/MS) was used to fingerprint a bioactive polylactic acid
P02-025
(PLA) with polybutylene succinate (PBS) (95:5) film extruded with
Mutagenicity evaluation by the Ames Test of
variable quantities of oregano essential oil. Bio-plastic pyrolysis
two organosulfur compounds
detected both, lactide enantiomers and monomer units from the
PLA and specific molecular markers from the PBS fraction. Oregano
M. Llana-Ruiz-Cabello 1 , A.I. Prieto 1 , S. Maisanaba 1 , S. Pichardo 1 ,
EO pyrolysis released aromatics and terpenes with cymene, ter-
E. Guillamón 2 , C. Nuñez 2 , A.M. Cameán 1,∗
pinene and thymol/carvacrol peaks identified as diagnostic peaks.
1 Linear correlation coefficients < 0.950R2 (p < 0.001) were found
Area of Toxicology, University of Sevilla, Sevilla, Spain
2 between the chromatographic area of these diagnostic peaks and
DOMCA S.A., Alhendín, Granada, Spain
the amount of oregano EO included in the bioplastic matrix. Our
results indicate that Py–GC/MS is a valuable tool providing not
In the food industry, one of the most promising use of Allium
only a precise fingerprinting but also precise information about
species is as antioxidant and antibacterial, since they are a natural
composition, quality and additives in active packages made with
alternative to chemical additives. In this sense, the incorporation
biogenic polymers.
of essential oil components, such as organosulphur compounds in
“active packaging” allows the controlled release of these active sub- Acknowledgements: Projects AGL2012-38357-C02-01 and
stances into the food, reducing the undesirable flavours caused in CGL2012-38655-C04-01, co-financed by FEDER. M Llana-Ruiz-
the case of the direct addition. For this reason, toxicological test- Cabello acknowledges Junta de Andalucia predoctoral grant
ing is needed to ensure the safety and quality control of Allium associated to AGR7252 project and NT Jiménez-Morillo to his FPI
species preparations used in food industry. The present work aims research grant (BES-2013-062573).
to perform for the first time a mutagenicity study of propilpropan-
http://dx.doi.org/10.1016/j.toxlet.2015.08.251
otiosulphonate (PTSO) and propilpropanotiosulphinate (PTS) by
Ames Test in the absence and presence of the S9 metabolic acti-
vation fraction. The incorporation version of the Ames test was P02-027
performed according to the recommendations of Maron and Ames Oxidative damage marker 8-OHdG in urine of
(1983) and following the principles of OECD guideline 471 (1997). vegetarians and omnivores
Five Salmonella typhimurium histidine-auxotrophic strains TA97A,
TA98, TA100, TA102 and TA104 were used for the assay. Five differ-
P. Koivisto 1,∗ , A.-L. Kosonen 2 , J. Laakso 3
ent concentrations of PTSO (1–50 ␮M) and PTS (8.75–140 ␮M) were
tested in three independent experiments. Each primary experiment 1 Evira, Chemistry and Toxicology, Helsinki, Finland
included one negative control (distilled water), one positive control 2 University of Eastern Finland, School of Applied Science and Teacher
for each strain (2-nitrofluorene, sodium azide or 2-aminofluorene)
Education, Kuopio, Finland
and one solvent control (dimethyl sulfoxide). Revertant colonies 3 Finnish Safety and Chemical Agency, Helsinki, Finland
were counted and background lawn was inspected for signs of
toxicity or compound precipitation. PTSO and PTS have shown
Introduction: Reactive oxygen species are harmful species but
mutagenic activity in TA98 strain only in presence of S9 metabolic
are formed through exercise and metabolism. Vegan diet may con-
 Abstracts / Toxicology Letters 238S (2015) S56–S383
tain more antioxidants than normal diet. In this study we analysed
urinary 8-OH-deoxyguaniosine – a marker of oxidation – from urine
of vegans and omnivores.
Materials and methods: Urine samples were collected from veg-
etarians and control subjects. Urine samples were purified with
solid phase extraction. Analysis of purified urine samples was made
with reversed phase UHPLC using methanol gradient. Detection of
8-OHdG was made with a triple stage quadrupole mass spectrom-
etry using multiple reaction monitoring using transmission value
284–168. Stable isotope was added to all samples as an internal
standard.
Results: Preliminary results indicate that level of 8-OHdG in
urine sample were low, but detectable in most samples. At group
level there was no statistical difference between vegans and omni-
vores.
http://dx.doi.org/10.1016/j.toxlet.2015.08.252
P02-028
Acute dietary risk assessment for the fungicide
carbendazim in fruits and vegetables consumed
in Brazil
P. Rembischevski 1,∗ , L. Nunes Gonçalves 1 , L.A. Nero 2
1 National Health Surveillance Agency – Anvisa, Toxicology Division,
Brasilia, Brazil
2 Universidade Federal de Viçosa, Departamento de Veterinária,
Viçosa, Brazil
The Brazilian pesticide market has reached near U$9 billion a
year in the last three years, ranking Brazil as the leading country
regarding the use of this substances in the world. In this context,
carbendazim is the 2nd most used fungicide and the 9th among
all classes of pesticides utilized in Brazil. Besides, carbendazim is
one of the three most detected pesticide in all violations observed
throughout the first decade of the Brazilian Programme for the
Analysis of Pesticide Residues in Food (PARA), the national moni-
toring programme, which covers 25 agriculture crops among fruits,
vegetables and cereals. Therefore, an evaluation of the dietary risk
to consumers was necessary, considering the toxicological impor-
tance of this substance due to its adverse reproductive effects
observed in experimental animals. The acute risk assessment is jus-
tified by the single dose effect of carbendazim in model animals,
besides its potential relevance in the mode of action for humans.
The recommended EFSA deterministic methodology was applied
and the results indicated that grape, lettuce, kale, orange, papaya,
and pineapple showed concern with regard to the acute intake risk
in some exposure scenario (general population or childbearing age
women), depending on which international Acute Reference Dose
(ARfD) was adopted: JMPR, EFSA, U.S. EPA, APVMA or CPMRA. For
example, the outcome for pineapple ranged from 44% of ARfD for
the general population if we consider the JMPR ARfD, to 2307%
of ARfD for women of childbearing age considering the CPMRA
ARfD, reflecting the huge discrepancy between the different bod-
ies in the establishment of this parameter. However, by refining
the results taking into account the fruits with no edible peel, only
orange and papaya exceeded some of the toxicological reference
parameters selected, although for pineapple it depended on the
Post Harvest Interval. In conclusion, attention must be driven to
the exceedance of the acute toxicological reference parameter for
carbendazim in some of the main foods in the diet of Brazilian
consumers. In the next future we intend to refine all these calcula-
tions by using the probabilistic approach, as well as performing the
chronic risk assessment for carbendazim along with thiophanate-
S73
methyl. This work also points out the importance of establishing a
national independent ARfD for this pesticide by Anvisa.
http://dx.doi.org/10.1016/j.toxlet.2015.08.253
P02-029
Efficient regulatory and safety assessment of
(new) food and feed products
S. van den Berg ∗ , C. van den Berg, W. Leeman, A. Kruizinga, H.
Heeres, L. Krul
TNO, Zeist, Netherlands
Food and feed has to comply with the applicable legislation that
is in place in the countries where they are marketed and above
all it has to be safe. The work required to meet the standards is
usually time-consuming, costly and often involves animal testing.
Moreover, safety assessment requires specialist expertise covering
multiple science domains. To prevent unnecessary research and
to provide guidance for an efficient regulatory and safety assess-
ment of (new) food and feed products, the Food Safety Assessment
Tool (FSAT) was developed. FSAT bridges product analysis and rel-
evant EU legislation with the science on hazard, exposure and risk
assessment. To provide up-to-date guidance on all different fields
relevant for regulatory and safety assessment of food and feed
products, FSAT contains decision trees (e.g. EU regulation, expo-
sure assessment, analytical methods), information sources like a
search engine to retrieve public toxicological data, an efficient
regulatory and safety assessment strategy to cover products and
mixtures, state-of-art knowledge on non-testing strategies (e.g.
TTC, read-across) contributing to a reduction of animal testing, and
a protocol for (further) toxicity testing strategies for all toxico-
logical endpoints to be used when data are insufficient for safety
substantiation of the product facilitating selection of the most opti-
mal research strategy. FSAT is developed as a web-based tool in
which the user can add relevant data following a standardized pro-
tocol and is thereby guided through each step via decision trees,
wizards, help boxes and specific tools. FSAT is being updated reg-
ularly. Although the current focus is on food safety, hence focused
on oral exposure, FSAT contains many elements making it also
applicable for risk assessment via other routes of exposure. Use
of FSAT increases transparency and efficiency of the assessment
process and decisions made. Moreover, FSAT provides the possi-
bility to easily share knowledge between experts and may even be
used for training purposes. All available product-related knowledge
and information can be integrated resulting in an optimal accessi-
bility to data available right from the start throughout the whole
research and/or assessment process. FSAT thus enables an efficient
assessment of EU regulatory compliance and food safety.
http://dx.doi.org/10.1016/j.toxlet.2015.08.254