Mister Chairman, Ladies and Gentlemen, Today I would like to give you some information about very important problem such as influence of food packages on the food quality.

MIGRATION FROM FOOD CONTACT MATERIALS The main source of contamination is a special layers used in order to protect the interior of the packaging from corrosion and to protect food from direct contact with the material used to produce the container. In the scientific literature, which publishes the latest research findings related to the monitoring of xenobiotics one can find increasingly more information on the sources and the volume of emissions released by everyday objects. One of the priority issues in this area of research is, without a doubt, the one of migration of low molecular weight compounds from the packaging to the food stored inside. 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

MIGRATION FROM FOOD CONTACT MATERIALS Bisphenol A Bisphenol A diglycidyl ether Bisphenol-A-monoglycidylether Bisphenol A(3-chloro-2-hydroxypropyl)glycidyl ether Bisphenol B Among the most frequently identified substances in simulants and canned food, there are: bisphenol A, bisphenol A diglycidyl ether (BADGE), its derivatives and compounds from the phthalates group. On the screen are presented chemical structures of some compounds Dibuthyl phtalate Bisphenol A dimethacrylate 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

MIGRATION FROM FOOD CONTACT MATERIALS Global migration - total mass of substances transferred from the packaging material to food during the time of their contact. Specific migration - mass of a single substance or a substance group transferred from the packaging material to food during the time of their contact. Compound SML [µg/kg foodstuffs or food simulants] BPA 600 Sum of BADGE and their hydrolysed derivatives 9000 Sum of BADGE and their hydrochloric derivatives 1000 A basic criterion for accepting food contact material are toxic properties of substances used to produce the material and the level of global migration and specific migration estimated on the basis of the value of tolerable daily intake (TDI). In Table there are presented limits for specific migration established by experts of the Scientific Committee for Food As it can be concluded from the data in the table the limit value have been established only for a small amount of compounds. In the light of that knowledge, it can be said that the food contact materials classification adopted nowadays raises many concerns 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

MIGRATION FROM FOOD CONTACT MATERIALS PROBLEMS AND HAZARDS: small number of identified contamination present in food products. All unidentified contamination is described as a NIAS (Non-Intentionally Added Substances), compounds may be transformed into various types of derivatives of physicochemical and toxicological properties different then the initial product, large number of compounds have similar properties to the compounds from EDC (Endocrine Disrupting Compounds) group Additionally, the situation is more complicated because, the number of identified synthetic compounds constitutes only a small part of all contamination present in food products. This situation raises seriously doubts as the fact that there is no possibility of identification and determination of all substances released from packaging. What is more, nowadays, it is known that as a result of technological processes and its interaction with food ingredients, migrating compounds may be subject to a transformation into various types of derivatives of physicochemical and toxicological properties different then the initial product. The lack of appropriate toxicological knowledge disable the possibility of assessing the real threat the consumer is subject to especially that the large number of compounds used to produce food contact material have similar properties to the compounds from the EDC group (Endocrine Disrupting Compounds). 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

Species of the test organism METHODOLOGY Test Species of the test organism Measured parameter Duration of the time Toxicity studied MICROTOX® Bacteria Vibrio fischeri Decrease in luminescence 30 min Acute toxicity OSTRACODTOXKIT F™ Ostracods Heterocypris Incongruens evaluation and assessment of mortality of organisms and inhibiting the growth 6 days Chronic toxicity In the light of this knowledge, it seems to the assessment of the real threat related to exposing a person to the xenobiotocs is possible only if an integrated approach is applied, based on the use of appropriately chosen bioassays. In the table are presented the most important information about applied bioassays 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

METHODOLOGY And now there are given some Information on experimental condition and sampling applied in research in order to extraction low molecular compounds from interior layer. Three stimulants of specific kinds of food and three different temperature have been chosen in relation to the methodologies used in order to assess global migration, as described in the EU Directives. 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

METHODOLOGY 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

METHODOLOGY 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

RESULTS The following slide presents results of measurements of acute and chronic toxicity obtained for samples, which used distilled water as a simulant medium. Taking into consideration the data obtained it can be concluded that temperature is the main factor, which influences intensification of a migration process. Exposure of bioindicators to samples that were subject to a heating process resulted in a significant inhibition of enzymatic activity of these organisms and nearly 90% reduction in bacterial luminescence and 30% inhibition of growth of crustaceans in comparison with the results obtained for samples stored at room temperature. 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

RESULTS The increase of bacterial luminescence as well as crustaceans growth observed for the samples taken after a short extraction time can be a result of the phenomenon of hormesis. 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

RESULTS [%] The following slide presents results of measurements for ethanol extract samples. These results indicate that biologically active compounds, which were present in all samples, cause a very high level of toxicity When comparing the results obtained for the samples taken from cans with a control sample, it can be observed that releasing an appropriate amount of xenobiotics that caused the ihibition of the bioluminescence occurred only after 24 hour extraction time (weakening of the luminescence by approx. 30%). 12 h 24 h 48 h 2 wk 12 h 24 h 48 h 2 wk 2 wk 2x 12 h 24 h 48 h 2 wk 2 wk 2x control Room temperature 65 oC 121 oC 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

RESULTS When using an acetic acid as the extraction medium it was possible to measure only acute toxicity, as long exposure of crustaceans to the acetic acid led to the death of all organisms. As you can see on the slide all the samples reveal very high levels of toxicity. Such a high level of inhibition of bioluminescence does not allow to determine the influence of temperature as well as storage time on the rate of release of toxic compounds from the internal surface of the package However, when comparing the result obtained for the control sample with other samples, it might be concluded that the biologically active compounds have been released to the model liquid. 24 h 48 h 2 wk 12 h 24 h 48 h 2 wk 2 wk 2x 12 h 24 h 48 h 2 wk 2 wk 2x control Room temperature 65 oC 121 oC 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

CONCLUSIONS Food Contact Materials have negative impact on the food quality Food Contact Material classification adopted nowadays does not allow to evaluate the actual impact of packaging on food stored inside Consumption of significant amounts of food stored in metal containers may pose a direct threat to consumer's health Based on the test results it can be concluded that as a result of contact between the inner layer of the packaging and model liquids the bound toxic ingredients of packaging used in the production of protective layers were washed out. In most cases dependless on the extraction time or temperature to which the samples were exposed the reduction of bioluminescence and growth inhibition was noted. Toxicological analysis of samples revealed that a system of evaluation of food contact materials, which is currently in use, does not allow to evaluate the actual impact of packaging on food stored in it. Based on the study results, it can be concluded that the adverse biological effect is caused by other substances, which are characterized by increased biological activity and which are not covered by monitoring when evaluating the quality of packaging material. The observed high level of toxicity of extract, indicates that consumption of significant amounts of food stored in metal containers may pose a direct threat to consumer's health 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia

THANK YOU FOR YOUR ATTENTION 6th International Conference and Exhibition on Analytical & Bioanalytical Techniques, 1-3.09.2015, Valencia