Non-Target Analysis Of Intentionally And Non Intentionally Added Substances From Plastic Packaging Materials And Their Migration Into Food Simulants

1Graphical abstract 2 1 1NON-TARGET ANALYSIS OF INTENTIONALLY AND NON INTENTIONALLY ADDED 2SUBSTANCES FROM PLASTIC PACKAGING MATERIALS AND THEIR MIGRATION INTO 3FOOD SIMULANTS 4 5 6V. García Ibarra**a, A. Rodríguez Bernaldo de Quirós*a, P. Paseiro Losadaa, R. Sendóna 7a Faculty of Pharmacy, Department of Analytical Chemistry, Nutrition and Food Science, 8University of Santiago de Compostela, Santiago de Compostela, 15782, Spain 9 10 *to whom correspondence should be addressed 11 12 e-mail: [email protected] +34 881814965. 13 **Present address: Faculty of Sciences and Food Engineering, Universidad Técnica de Ambato, 14 Campus Huachi, Ambato, Ecuador. 15 e-mail: [email protected] 16 17 18 19 20 21 22 23 2 24 25 26 ABSTRACT 27 A wide variety of compounds are used in the manufacture of plastic materials; however, some 28 of them have the potential to migrate in food products. 29 The main objective of this work was the identification of potential migrants in plastic 30 packaging materials and their migration into food simulants. For this purpose, a total of twelve 31 plastic packaging materials were analyzed using purge and trap (P&T) coupled to GC–MS for 32 volatile compounds and GC-MS for semivolatile compounds after extraction with organic solvents. 33 A qualitative migration tests were carried out using tenax and isooctane and the migrants were 34 analyzed by GC-MS. About 100 different organic compounds were detected in plastic materials. Of 35 these, 27 compounds migrated into tenax and/or isooctane. In general, migration occurs to a larger 36 extent in tenax than in isooctane. Moreover, most of detected compounds in plastic samples are not 37 included in the Commission Regulation (EU) No. 10/2011 on plastic materials and articles intended 38 to come into contact with food. 39 40 Keywords: volatile compounds; semivolatile compounds; migration; food simulants 3 41 1. Introduction 42 Plastic materials are widely used in food packaging applications for their excellent properties. 43 Polymers most widely used in this application include polyethylene (PE), polyethylene 44 terephthalate (PET), polypropylene (PP), polyamide (PA), polystyrene (PS), ethylene vinyl alcohol 45 (EVOH), polyvinylidene chloride (PVDC), polyvinyl chloride (PVC), ethylene vinyl acetate (EVA) 46 and polycarbonate (PC). The mentioned polymers are also used too to produce multilayer materials 47 (Hoppe, de Voogt, & Franz, 2016). Multilayer materials are commonly used in food packaging 48 industry. These materials are produced by combination of various types of plastic and or non-plastic 49 materials (e.g., aluminum foil and paper, among others) with adhesives. Most commonly used 50 adhesives in the manufacturing of multilayer food packaging are polyurethane and acrylic or 51 acrylate based adhesives. The final properties of these materials are a combination of individual 52 material properties, which allows improving the performance of packaging materials (Clemente, 53 Aznar, Nerín, & Bosetti, 2016). 54 Packaging provides physical protection and extends the shelf life of foods, however when 55 packaging comes in contact with food, components from the packaging materials can migrate into 56 the food. there is the possibility that constituents from the same are transferred to the packaged 57 food. In fact, packaging materials contain chemical compounds that potentially can migrate into the 58 foodstuffs; tThese substances could include authorized substances e.g. residual monomers, starting 59 substances as well additives used to improve the properties of materials. Furthermore, may migrate 60 the so called “non intentionally added substances” or (NIAS). from packaging material into the 61 packaged food. In Europe the substances authorized are included in Regulation N° 10/2011 which 62 contains the list of monomers, starting substances and additives allowed to be used in the 63 manufacture of plastics as well as the list of simulants and the assay conditions to be used for 64 testing migration. of constituents of plastic materials and articles intended to come into contact with 65 foodstuffs. The same Regulation defines NIAS as “any impurities in the substance used or reaction 66 intermediates formed during the production process or decomposition or reaction products”. These 4 67 substances are not included in the union list, however the safety of NIAS has to be assessed 68 (Commission Regulation (EU) No 10/2011). 69 The presence of NIAS in packaged food could be the result of the impurities present in the raw 70 materials used for their production but one of the most frequent pathway to NIAS formation is 71 degradation processes (Dąrowska, Borcz, & Nawrocki, 2003; Kim et al., 2015). For example, 72 during PET manufacturing, several degradation and decomposition reactions can occur. High 73 temperatures and the presence of oxygen in the PET can promote reactions generating numerous 74 NIAS in the polymer (Kassouf, Maalouly, Chebib, Rutledge, & Ducruet, 2013). In recent years the 75 concern about the risk of NIAS is increasing. Over 50% of compounds migrating from food contact 76 materials are NIAS (Bach et al., 2013; Grob, Biedermann, Scherbaum, Roth, & Rieger, 2006). From 77 the toxicological point of view, particular attention have received low molecular substances, 78 including also NIAS, given that it is generally accepted that substances with molecular weight over 79 1000 Da are not absorbed by the gastro-intestinal tract (European Food Safety Authority, 2008). 80 Several analytical techniques have been used for the determination of potential migrants in 81 packaging materials used target and non target methodologies. The application of mass 82 spectrometry (MS) in combination with gas chromatography (GC) or liquid chromatography (LC) 83 has been well recognized for both quantification and semi-quantitative screening of food packaging 84 materials. GC is suitable for semi-volatile substances, whereas compounds that are thermally 85 instable, non- or highly volatile should be analyzed by LC (Bradley & Coulier, 2007; Lahimer, 86 Ayed, Horriche, & Belgaied, 2013; Nerin, Alfaro, Aznar, & Domeño, 2013; Simal-Gándara, 87 Damant, & Castle, 2002). 88 GC-MS coupled to headspace by solid-phase microextraction (HS-SPME) or combined with a 89 purge & trap (P&T) system have shown to be efficient techniques for screening of volatile 90 compounds in food contact materials (Nerin, Canellas, Aznar, & Silcock, 2009; Skjevrak et al. 91 2005). More sophisticated technologies such as comprehensive two-dimensional gas 5 92 chromatography has also been successfully applied in the analysis of food packaging materials 93 (Biedermann, Castillo, Riquet, & Grob, 2014; Biedermann, & Grob, 2013). 94 Other techniques such as pyrolysis–GC coupled to mass spectrometry has also been used for 95 compositional analysis of polymers (Rial-Otero, Galesio, Capelo, & Simal-Gándara, 2009). 96 Gas chromatography-mass spectrometry (GC-MS) has been applied successfully for the screening 97 and the identification of potential migrants from films and adhesives as well as for quantitative 98 analysis of migration into food simulants. Rothenbacher & Schwack (2009) developed a GC-MS 99 method for the screening in resins, foils, and multilayer foils intended for the production of food 100 packaging. Toxicological evaluation of the 46 identified compounds was also realized. GC-MS 101 coupled to headspace by solid-phase microextraction (HS-SPME) is another technique selected for 102 its high sensitivity for the screening of volatile compounds coming from adhesives used in 103 packaging materials (Nerin, Canellas, Aznar, & Silcock, 2009). 104 Other technique that can trap and analyze simultaneously the vapor emitted from the plastic is the 105 purge & trap (P&T) system coupled with a GC-MS. This system allows the identification and 106 quantification of compounds that arriving at the MS detector. Skjevrak et al. (2005) used Purge- 107 and-trap (P&T) combined with GC-MS (P&T-GC/MS) and solid phase extraction (SPE)-GC/MS 108 for the analysis of volatile and semi-volatile compounds originated from four type of plastic 109 articles. Most migrants were NIAS or compounds originated from non-plastic components, such as 110 printing inks, adhesives, not-listed additives, solvents and coatings. 111 Advances in analytical instrumentation have allowed to address the challenging task to identify 112 unknowns. High resolution mass spectrometry techniques such as Time of Flight (TOF) and 113 Orbitrap have been effectively applied for this purpose (Isella, Canellas, Bosetti, & Nerin, 2013; 114 Cherta et al., 2015; Vaclavikova et al., 2016). 115 the use of new technologies to identify target and non-target compounds such as Time of Flight 116 (TOF) mass spectrometry, which provides the sensitivity and selectivity required for the analysis of 117 compounds with an accurate mass analysis. Ultra-high performance liquid chromatography– 6 118 quadrupole time-of-flight–mass spectrometry (UPLC-Q-TOF/MS) has been proved to be a powerful 119 technique to identify NIAS from polyurethane adhesives (Isella, Canellas, Bosetti, & Nerin, 2013). 120 In the same way the identification of unknown substances capable to migrate from plastic materials 121 to food simulants was performed using the combination of two analytical techniques (GC– 122 (EI)TOF/MS) and GC-(APCI)QTOF/MS, which allowed the confirmation of the identity of 8 123 migrants (Cherta et al., 2015). Recent developments have let that polymers and solid food simulants 124 can be analyzed by direct thermal desorption techniques, in which the extraction steps can be 125 avoided. These techniques include direct analysis in-real-time (DART) MS (Ackerman, Noonan, & 126 Begley, 2009; Lago & Ackerman, 2016), desorption electrospray ionization (DESI) MS (Venter, 127 Nefliu, & Cooks, 2008) and atmospheric solids analysis probe (ASAP) MS (Barrere, Maire, 128 Afonso, & Giusti, 2012). 129 130 In this work an approach based on purge and trap and MS (P&T-GC/MS) and an extraction with 131 organic solvents followed by a GC-MS analysis for the determination of volatile and semi-volatile 132 Intentionally added substances (IAS) and non-intentionally added substances (NIAS) in plastic 133 packaging materials is proposed. This methodology could be a valuable screening tool and be used 134 as a first step in identifying potential migrants for compliance with food contact materials 135 legislation. Additionally, migration tests were performed using Tenax and isooctane as food 136 simulants.In this work a GC-MS and purge and trap (P&T-GC/MS) method was developed for the 137 determination of volatile compounds in twelve different plastic food-packaging samples. In 138 addition, the semi-volatile compounds were analyzed by GC-MS after an extraction with organic 139 solvents. Migration tests were performed with the aim of identifying the semi-volatile compounds 140 capable to migrate from plastic materials into food simulants Tenax and isooctane. 7 141 2. Materials and methods 142 2.1. Chemicals and analytical standards 143 Analytical standards of butylated hydroxytoluene (BHT) 99% (CAS: 128-37-0), tributyl acetyl 144 citrate (ATBC) 99% (CAS: 77-90-7), bis(2-ethylhexyl) adipate (CAS: 103-23-1), bis (2- 145 ethylhexyl) phthalate (DEHP) 99% (CAS: 117-81-7) were purchased by Fluka (Steinheim, 146 Germany). 147 Caprolactam (CAS: 105-60-2), diethyl phthalate (DEP) 99.5 %, (CAS: 84-66-2), erucamide (CAS: 148 112-84-5), tolulene 2,6-diisocyanate 97% (CAS: 91-08-7), benzophenone 99%, internal standard 149 methyl myristate (CAS: 124-10-7), saturated alkane standard mixture C7-C30 (1000 μg/mL each 150 component in hexane) and polyoxide 2,6-diphenyl-p-phenylene TA (Tenax), 80-100 mesh were 151 purchased from Sigma Aldrich (Schnelldorf, Germany). Toluene 2,4 diisocyanate (584-84-9) from 152 Merk (Darmstadt, Germany). 153 All chemicals were of analytical grade. Acetonitrile (ACN), ethanol, methanol, acetone and 154 isooctane were from Merck (Darmstadt, Germany). Purified water (Type I) was obtained from an 155 Autwomatic Plus purification system (Wasserlab, Navarra, Spain). 156 2.2. Samples 157 Twelve samples of plastic films (with and without printing ink) intended to come into contact with 158 food not yet in contact with food (six samples purchased in Quito-Ecuador and six samples 159 purchased in Spain) were independently studied during the investigation of organic compounds. 160 Details of packaging samples are listed in Table 1. 161 2.3. Sample Treatment (Screening) 162 The organic compounds originated from the plastic material, were extracted from slices of plastic 163 material (approx. 1 g, 1.58 dm2) by purging an inert gas (Helium at 40 mL/min) through the sample 164 (heated at 60ºC) during 20 minutes. Purged sample components are trapped in a tube containing a 165 suitable sorbent material (Vocarb 3000 Trap). When purging is completed, the trap is heated 8 166 (250ºC) and back flushed with Helium (400 mL/min during 2 min) to desorb the trapped sample 167 components into the GC column. Components eluting from the GC column are identified by MS. 168 A liquid extraction was also performed with plastic samples. Acetonitrile was selected as extraction 169 solvent due to its extraction power. For this purpose an An area of 0.8 dm2 (0.7 g) was cut from 170 each packaging material and analysed separately. Each specimen was then cut in layers, pieces put 171 in a glass container and extracted with 25 mL of acetonitrile for 24 h at 70 °C, after been 172 hermetically closed. An aliquot of the extract (10 mL) was then removed and evaporated under 173 nitrogen stream (RapidVap Vertex Evaporator, Labconco) up to less than 1 ml. Subsequently, to a 174 1-mL volumetric flask was added 50 μl of the internal standard methyl miristate (100 mg/L) and the 175 volume was made up with the final extract (evaporated). 176 Cramer rules were used to estimate the toxicity of identified compounds. An open source 177 application called Toxtree v2.6.13 (Ideaconsult Ltd.) was used for this purpose (Toxtree v2.6.13, 178 2015). The decision tree allows classifying chemical compounds into three structural classes mainly 179 on the basis of chemical structure and reactivity. Cass I (low toxicity): For substances with simple 180 chemical structures and efficient modes of metabolism; Class II (intermediate toxicity) for 181 substances that may suggest significant toxicity or have reactive functional groups and substances 182 of a chemical structure that permit no strong initial impression of safety and may even suggest a 183 significant toxicity are classified in Class III (high toxicity) (Lapenna & Worth, 2011; Patlewicz, 184 Jeliazkova, Safford, Worth, & Aleksiev, 2008). 185 2.4. Qualitative Migration Test 186 Migration test were carried out using two simulants. Isooctane and Tenax® . All migration 187 experiments were done in duplicate. 188 For migration test performed with Tenax, 0.13 dm2 of each laminate (the side in contact with food) 189 were placed in a petri dish and covered with 0.52 g of Tenax (4 g Tenax per dm2 laminate) (UNE- 190 EN-14338, 2004) which had been previously cleaned with acetone in a Soxhlet extraction for 6 h 191 and dried into the oven for other 6 h at 160°C. 15 344 DEHA exposure is described in the literature. Fromme et al. (2007) reported values of daily intake 345 via food of 0.7 μg/kg body weight for DEHA. 346 Regarding phosphates, triphenyl phosphate (TPP) was detected in samples 3EC and 9EC. This 347 compound is used as plasticizer for cellulose acetate articles and in lacquers, as well as a flame 348 retardant and solvent (Sheftel, 2000). This compound classified in class III according Cramer rules. 349 Tributyl aconitate (TBA), a compound commonly used to protect PVC against effects of light and 350 heat, was detected in samples 6ES, 7ES, 8ES and 12ES (Dupáková, Dobiáš, Votavová, Klaudisová, 351 & Voldrich, 2010). Isopropyl myristate was detected in sample 3EC, and is used as plasticizer for 352 cellulosic, as well as pigment dispersant and binder (Ash & Ash, 2004). 353 Antioxidants such as BHT were detected in samples of PP/PET, PE/EVA and PE/PET (8ES, 10ES, 354 11ES and 12ES). This compound is used in the production of plastics, mainly polyolefins and is 355 approved as antioxidant for food contact applications (Sheftel, 2000). 356 Slip agents were detected in all types of materials analyzed PE/PET, PP/PET, PE/PA, PE/PP and 357 PE/EVA. According to the bibliography these compounds are added to reduce the surface 358 coefficient of friction of polymers and are used to enhance either processing or end applications. 359 Erucamide was found in all tested samples, oleamide in six samples, hexadecanamide in three 360 samples and octadecanamide in two samples. Octadecamide is used as a slip agent, anti-fogging or 361 lubricant for food packaging films (mainly polyolefins) (Ash, 2004). Erucamide is one of the most 362 commercially important slip additives, used in polyolefin closures as well as in food contact 363 materials due to their properties including heat stability as well as better blocking performance 364 (Lahimer et al., 2013; Wypych, 2005). Hexadecanamide is used as slip agent for processed plastics, 365 printing inks, coatings and films (Dupáková et al., 2010). Oleamide is used as slip agent for printing 366 inks, coatings and films however erucamide is more widely used because it is thermally more stable 367 than oleamide (Dupáková et al., 2010, Kumudini, 2012; Wypych, 2005). 368 Caprolactam was detected in PE/PA materials (samples 5EC and 9EC) and in PE/PET materials 369 (samples 2EC and 4EC). In these last samples probably polyamide (PA) was present in one of the 16 370 inner layers of the packaging materials. PA in most cases is combined with polyolefins as a 371 component of multilayer structure. Caprolactam is a monomer used for the manufacture of 372 polycaprolactam (nylon 6), widely used in food packaging materials as well as in the manufacture 373 of printing inks (Pogorzelska & Mielniczuk, 2001; Sheftel, 2000). According to IARC caprolactam 374 is probably not carcinogenic to humans (Group 4) and in experimental animals there is evidence 375 suggesting a lack of carcinogenicity of this compound (International Agency for Research on 376 Cancer,1999). Other compounds of high toxicity, identified in our samples were diisocyanates, 377 which are chemical compounds mainly used in polyurethane applications, such as coatings, 378 adhesives, elastomers, sealants among others (Dupáková et al., 2010). 2,4-toluene diisocyanate was 379 identified in 3 samples (7ES,8ES, 12ES) while 2,6-Toluene diisocyanate in sample 7ES. Their use 380 in food packaging materials is regulated in the European Union, however their levels in final 381 products must not exceed 1.0 mg/kg (Commission Regulation (EU) No 10/2011). The photoinitiator 382 benzophenone was detected in samples 7ES and 12ES, the origin of this compound in tested 383 samples is possibly the printing inks used in the external face of packaging films. This compound 384 widely used as initiator for printing inks cured by UV radiation is possibly carcinogenic to humans. 385 (International Agency for Research on Cancer, 2013). 386 The compounds benzenesulfonamide, n-ethyl-2-methyl- and benzenesulfonamide, n-ethyl-4-methyl 387 were also detected in two samples (3EC and 9EC). These compounds have been used as component 388 of ink formulations and are classified in class III of toxicity according to Cramer rules. In the 389 European Union, these substances are not included on the list of authorized substances in the 390 manufacture of plastic materials and articles. HoweverHowever, the U.S. Food and Drug 391 Administration (FDA) listed these substances in the Inventory of Effective Food Contact 392 Substances (FCS) Notifications. (U.S Food & Drug Administration, 2017). 393 Glycols such as triethylene glycol and tripropylene glycol were also identified. Triethylene glicol, a 394 compound widely used as solvent in coating and ink industry, as well as a solvent for nitrocellulose 395 and various resins was detected in sample 12ES (PE/PET) (Lago & Ackerman, 2016). Tripropylene 17 396 Glycol was detected in sample 8ES and is used as initiator for urethane polyols, in the manufacture 397 of polyester plastics as well as solvent and homogenizing agent for inks. Synthetic fatty alcohols 398 such as methyl palmitate was detected in sample 7ES (PE/EVA). This compound is used as 399 intermediate for detergents, emulsifiers, stabilizers, resins, lubricant, plasticizers and defoamer in 400 food-contact coatings (Ash & Ash, 2004). 401 Other identified compounds can be “non-intentionally added substances” (NIAS), such is the case 402 of 2,4-di-tert-butylphenol and 2,6-di-tert-butyl-1,4-benzoquinone. 2,4-di-tert-butylphenol is a 403 degradation product of Irgafos 168 while 2,6-di-tert-butyl-1,4-benzoquinone is a degradation 404 product of antioxidants such as Irganox 1010, Irgafos 168 and Irganox PS 802 (Félix, Isella, Bosetti 405 & Nerín, 2012; Lago &Ackerman, 2016). These compounds were detected in five and three 406 samples respectively and are reported as NIAS in other migration studies (Alin & Hakkarainen, 407 2011; Félix et al., 2012; Lago & Ackerman, 2016). 408 409 The compound tert-butyl-1-oxaspiro (4,5) deca-6-9-diene-2,8-dione, a byproduct of the antioxidant 410 Irganox 1010 was identified in almost all samples of PE/PET, PP/PP, PE/PA, LDPE/PP and 411 PE/PVDC (1EC, 3EC, 4EC, 5EC, 6ES, 7EC, 8EC, 9EC, 10ES, 11ES, 12ES). This compound 412 exhibited high toxicity according to Cramer rules and other authors have also estimated some 413 toxicity (Lago & Ackerman, 2016; Rothenbacher & Schwack, 2009). 414 415 Squalene was identified in samples of PE/PET, PE/PA, PE/PP and PE/ EVA (2EC, 4EC, 5EC, 6ES, 416 7ES and 12ES). This compound is an ethylenic-unsaturated hydrocarbon that had oxygen- 417 scavenging capacity to extend the shelf life of oxygen sensitive products. This compound may 418 bemay be added to packaging materials such as polyethylene and polypropylene during 419 conventional mixing processes to thermoplastics (Lopez-Rubio et al., 2004). 420 Lubricants such as glycerol trycaprilate were found in sample of PE/PET (2EC). Lubricants are 421 used to minimize adhesion and viscosity of plastic materials. 18 422 Some compounds were identified only in samples from Ecuador, such as caprolactam, 423 benzenesulfonamide, n-ethyl-2-methyl-, benzenesulfonamide, n-ethyl-4-methyl, isopropyl myristate 424 and octadecanal. Others such as BHT and DEHA only in samples from Spain. Scientific 425 information of some compounds listed in Table 4 was not available. Nevertheless, these results 426 provide evidence of the wide range of compounds that can be found in plastics materials intended 427 for food contact, which may be potential migrants to foodstuffs. Most of the identified compounds 428 are not included in the positive list of Commission Regulation (EU) No 10/2011 on plastic materials 429 and articles intended to come into contact with food, and only 3 of the compounds detailed in Table 430 3 and 16 of the compounds detailed in Table 4 are regulated by these normative. 431 It is interesting to note that some of the compounds identified are included in the inventory list of 432 the European Printing Ink Association (EuPIAincluir ref), in different categories like this octane, 433 undecane and tetradecane are within the solvents; hexane, dodecane, octadecane, hexadecane 434 diethyl phthalate, 2,4-di-tert-butylphenol, isopropyl myristate, diisobutyl phthalate, methyl 435 palmitate and 1-docosanol are included in substances used as additives or ingredients of additive 436 preparations; 1-tetradecanol is included in polymeric resin-monomers/precursors/raw materials; 1- 437 Octadecanol and triphenyl phosphate belong to pigment additives; and benzenesulfonamide, N- 438 ethyl-2-methyl and benzenesulfonamide, N-ethyl-4-methyl are included in both groups substances 439 used as additives or ingredients of additive preparations and polymeric resin - 440 monomers/precursors/raw materials. 441 442 3.2 Estimation of migration into food simulants 443 Considering the large number of compounds detected in the screening of plastic packaging samples, 444 a qualitative migration test was performed to identify the potential of all of these previously 445 identified substances to migrate into food simulants. 446 Qualitative migration tests were carried out with 12 plastic packaging materials using tenax and 447 isooctane as food simulants in contact with the plastic materials. Migration was performed from one 19 448 side of each laminate (side of contact with the food) and two independent replicates of each 449 packaging material were analyzed. The acetone extracts of tenax and isooctane were analyzed by 450 GC-MS with the same conditions used in the identification of semivolatile compounds. Results are 451 listed in Table 5 and Table 6. In total 27 compounds were detected in the migration experiments, of 452 which, 25 compounds migrated from samples from Ecuador while 17 migrated from samples from 453 Spain. Some of them were common in both simulants and others were only observed in one of 454 them. 455 In this work we have proposed an approximation with the purpose of having an idea about To 456 evaluate the differences in the amount of compound present in plastic materials after extraction with 457 acetonitrile and in migration into tenax and isooctane, . The responses were calibrated against 458 estimated concentrations of these compounds were obtained considering the response factor of the 459 internal standard methyl myristate, as reported in Equation 1 (Guazzotti, Giussani, Piergiovanni, & 460 Limbo, 2015; Jenke & Odufu, 2012). 461 462 463 (1) 𝐶 𝑎 = 𝐶 𝑖 𝑥 ( A a A i ) 464 Where Ca is the analyte concentration (mg/L), Ci the internal standard concentration (mg/L), Aa 465 the analyte area and Ai the internal standard area. 466 Migration test were performed in two food simulants, tenax and isooctane for dry and fatty food 467 products, respectively. Migration of caprolactam occurred from the samples 2EC, 4EC and 5EC 468 only into tenax, while in isooctane was not detected. It is important to note that caprolactam that 469 was present in an external layer (polyamide) of a packaging material 5EC and 9EC, could migrate 470 through the layers of packaging and reach the food simulant. 471 2,6-di-tert-butyl-1,4-benzoquinone migrates in tenax from samples PP/PP, PE/EVA and PE/PET 472 (3EC, 7ES, 10ES) in concentrations from 0.00045 to 0.0018 mg/dm2. 2,4-di-tert-butylphenol 473 migrate from five packaging samples of PP/PP (3EC), PE/PET (4EC, 12ES) and PE/PVDC 20 474 (10ES,11ES) into both simulants tenax and isooctane, with higher concentrations in tenax than 475 isooctane in most of the samples. In the case of samples 3EC, 10ES and 12ES, migration of 2,4-di- 476 tert-butylphenol was lower in isooctane than in tenax. In the sample 11ES migration value was 477 similar in both simulants, while in the sample 4EC migration was higher in isooctane than in tenax. 478 Our values were lower than those reported by Félix et al. (2012), except the samples 10ES and 479 11ES whose values were similar to those described by these authors. 480 BHT migrates only into tenax from the sample of PE/PET (12ES) in a concentration of 0.000761 481 mg/dm2, while migration was not observed in the samples of PP/PET, PE/EVA and PE/PET. For 482 this compound, the migration value obtained was lower than those reported by Félixby Félix et al. 483 (2012). 484 In terms of phthalates, DEP migrated in both simulants in six samples (3EC, 5EC, 6ES, 7ES, 8ES 485 and 9EC). Its migration was higher into isooctane in samples 3EC, 6ES, 7ES and 8ES, while in 486 sample 5EC the migration was higher in tenax. Migration was almost in the same proportion in both 487 simulants in sample 9EC. Migration values in isooctane were from 0.0012 to 0.084 mg/dm2, while 488 in tenax were from 0.0010 to 0.017 mg/dm2. Diisobutyl phthalate migrated from five samples 4EC, 489 5EC, 7EC, 11ES and 12ES into tenax and isooctane. Its migration was similar in both simulants in 490 samples 7ES and 11 ES, while in the rest of the samples migration values were higher in isooctane 491 than in tenax. Bis (2-ethylhexyl) phthalate migrated from the samples 2EC, 3EC, 9EC and 11ES in 492 tenax and isooctane, with highest migration values in sample 3EC in both simulants. 493 Benzenesulfonamide, N-ethyl-2-methyl- and benzenesulfonamide,n-ethyl-4-methyl migrated into 494 samples 3EC (PP/PP) and 9EC (PE/PA). Migration of benzenesulfonamide,n-ethyl-4-methyl was 495 higher in tenax than isooctane in both packaging materials. Benzenesulfonamide, n-ethyl-2-methyl 496 migrated more in tenax than isooctane in the sample 3EC, while in the case of sample 9EC 497 migration ranged in a similar level into both simulants. 498 Isopropyl myristate migrated only in the sample 3EC in both simulants, with higher migration 499 values in tenax than isooctane. 1-Hexadecanol migrated only into tenax from samples 3EC and 7ES, 21 500 with values from 0.0028 to 0.0084 mg/dm2. Octadecanal migrated into both simulants from sample 501 3EC. Its migration into isooctane was higher than in tenax. Acetyl tributyl citrate was identified in 502 different types of packaging materials including PP/PP, PE/PET, PE/PA, PE/PP and PE/EVA films 503 in levels up to 2.0 mg/dm2. This compound migrated from samples 3EC, 6ES, 7ES, 8ES, 9EC, 504 10ES, 11ES and 12ES into tenax and isooctane. In samples 3EC and 12ES migration was higher in 505 isooctane while in sample 6ES migration values were similar in both simulants. In the rest of 506 samples migration was greater in tenax than isooctane. ATBC was one of the predominant migrant 507 in both simulants, with migration values from 0.00026 to 0.96 mg/dm2 in isooctane and from 0.0013 508 to 0.58 mg/dm2 in tenax. The highest concentration level was observed in isooctane. 509 Other plasticizers found in migration studies were triphenyl phosphate (TPP) and tributyl aconitate. 510 TPP migrated from packaging samples 3EC and 9EC into both simulants, with higher migration 511 values in tenax. Migration of tributyl aconitate into both simulants occurred from samples 6ES, 512 7ES, 8ES, 9EC and 12ES, with higher migration values in tenax than isooctane in all samples, 513 except in the case of sample 8ES where the migration in isooctane was greater than in tenax. 514 Another compound also relevant in both simulants was 7,9 di-tert-butyl-1-oxaspiro (4,5) deca-6-9- 515 diene-2,8-dione, that migrate in eleven of twelve tested samples. Migration was higher in tenax than 516 isooctane, except in the case of samples 7ES and 11 ES when migration values are similar in both 517 simulants. Migration values in isooctane were from 0.00075 to 0.013 mg/dm2, whereas in tenax 518 from 0.0011 mg/dm2 to 0.046 mg/dm2. Oleamide migrated in sample 3EC in both simulants. The 519 highest concentration level was observed in tenax. 520 DEHA was found in three of twelve tested plastic materials in concentrations from 0.024 to 1.0 521 mg/dm2 and migrated in both simulants in the three samples of PE/PVDC and PE/PET (10ES, 11ES 522 and 12ES). Its migration was higher into tenax than into isooctane, migration values from 0.023 to 523 0.081 mg/dm2 were reached in tenax. The slip agent erucamide was detected in all plastic materials 524 with concentrations from 0.082 to 1.3 mg/dm2. In the same way this compound migrated in all 525 tested samples in tenax and isooctane, with concentrations from 0.0011 to 0.14 mg/dm2 and from 22 526 0.0096 to 0.19 mg/dm2 in isooctane and tenax, respectively. Migration values were higher in tenax 527 than in isooctane in all cases. Hexadecanamide migrated only in samples 3EC and 9EC, with higher 528 migration values in tenax than in isooctane. Octadecanamide migrated from PE/PA film (9EC) in 529 tenax and isooctane. Its migration into tenax was greater than in isooctane. 530 Squalene migrated into tenax and isooctane from samples 2EC, 4EC, 5EC, 6ES, 7ES and 12E 531 (PE/PET, PE/PA, PE/PP and PE/EVA), reaching high concentrations in tenax in almost all samples 532 tested. Migration of alkanes such as tetradecane, hexadecane, heptadecane, octadecane and 533 dodecane was also occurred in some samples, with higher migration values into tenax than in 534 isooctane. In general migration results showed higher migration values in tenax than in isooctane. 535 Only nine of out of all migrant compounds appeared in the positive list of Regulation 10/2011. 536 4. Conclusions 537 In this work the determination of volatile and semivolatile compounds in 12 packaging materials 538 were performed using P&T coupled to GC-MS for volatile compounds and GC-MS for semivolatile 539 compounds. These techniques have demonstrated to be useful for screening and identification of 540 compounds from plastic materials as well as their migration into food simulants. About 100 volatile 541 and semivolatile compounds including “non intentionally added substances” were detected in 542 plastic materials. Of these, 27 compounds migrated into tenax and/or isooctane. In general, 543 migration occurs to a larger extent in tenax than in isooctane. 544 On the other hand, it is interesting to point out that most of detected compounds and even more, the 545 migrants detected in the food simulants tested are not included in the positive list of monomers and 546 additives that are allowed to be used in plastic food contact materials; and neither in the . inventory 547 list of the European Printing Ink Association. 548 549 Conflict of interest statement 550 The authors declare that there are no conflicts of interest. 551 23 552 Acknowledgments 553 554 The study was financially supported by the Ministerio de Economía y Competitividad, Fondo 555 Europeo de Desarrollo Regional (FEDER), and Agencia Estatal de Investigación, Ref.No. 556 AGL2015-69609-P “MIGRAEXPO” (MINECO/FEDER, UE). V. García Ibarra was supported by a 557 graduate fellowship from the Secretaría Nacional de Educación Superior, Ciencia, Tecnología e 558 Innovación of Ecuador (SENESCYT). References Alin, J., & Hakkarainen, M. (2011). Microwave heating causes rapid degradation of antioxidants in polypropylene packaging, leading to greatly increased specific migration to food simulants as shown by ESI-MS and GC-MS. Journal of Agricultural and Food Chemistry, 59, 5418-5427. http://doi.org/10.1021/jf1048639 24 Ash, M. (2004). Handbook of green chemicals. (2nd ed.). New York: Endicott, Synapse Information Resources. Ash, M., & Ash, I. (2004). Handbook of Preservatives. New York: Synapse Information Resources. Ackerman, L. K., Noonan, G. O., & Begley, T. H. (2009). Assessing direct analysis in real-time- mass spectrometry (DART-MS) for the rapid identification of additives in food packaging. Food Additives & Contaminants, 26, 1611-1618. http://doi.org/10.1080/02652030903232753 Bach, C., Dauchy, X., Severin, I., Munoz, J.F., Etienne, S., & Chagnon, M.C.(2013). Effect of temperature on the release of intentionally and non-intentionally added substances from polyethylene terephthalate (PET) bottles into water: chemical analysis and potential toxicity. Food Chemistry, 139, 672-680. http://doi.org/10.1016/j.foodchem.2013.01.046 Barrere, C., Maire, F., Afonso, C., & Giusti, P. (2012). Atmospheric solid analysis probe-ion mobility mass spectrometry of polypropylene. Analytical Chemistry, 84, 9349-54. http://doi.org/10.1021/ac302109q Bengtstrom, L. (2014). Chemical identification of contaminants in paper and board food contact materials. (PHD Thesis). Available from: https://www.google.es/?gfe_rd=cr&ei=MAIwWfTTIrSp8wfpk6DYBg&gws_rd=ssl#q=Chemical+i dentification+of+contaminants+in+paper+and+board+food+contact+materials Biedermann, M., & Grob, K. (2013). Assurance of safety of recycled paperboard for food packaging through comprehensive analysis of potential migrants is unrealistic. Journal of Chromatography A, 1293, 107– 119. http://dx.doi.org/10.1016/j.chroma.2013.04.009 Biedermann, M., Castillo, R., Riquet, A. M., & Grob, K. (2014). Comprehensive two-dimensional gas chromatography for determining the effect of electron beam treatment of polypropylene used for food packaging. Polymer Degradation and Stability, 99, 262-273. http://dx.doi.org/10.1016/j.polymdegradstab.2013.10.021 Bradley, E., & Coulier, L. (2007). An Investigation into the Reaction and Breakdown Products from Starting Substances Used to Produce Food Contact Plastics, Report FD07/01, London (UK): Central Science Laboratory. Bush, J., Gilbert, J., & Goenaga, X. (1993). Spectra for the identification of monomers in food packaging (Vol. 14515). London: Springer Science & Business Media. Cao, X. L. (2010). Phthalate esters in foods: sources, occurrence, and analytical methods. Comprehensive Reviews in Food Science and Food Safety, 9, 21-43. Cherta, L., Portolés, T., Pitarch, E., Beltran, J., López, F. J., Calatayud, C., Company, B., & Hernández, F. (2015). Analytical strategy based on the combination of gas chromatography coupled to time-of-flight and hybrid quadrupole time-of-flight mass analyzers for non-target analysis in food packaging. Food Chemistry, 188, 301-308. http://doi.org/10.1016/j.foodchem.2015.04.141 Clemente, I., Aznar, M., Nerín, C., & Bosetti, O. (2016). Migration from printing inks in multilayer food packaging materials by GC-MS analysis and pattern recognition with chemometrics. Food Additives & Contaminants, Part A, 33, 703-714. http://doi.org/10.1080/19440049.2016.1155757 Commission Regulation (EU) No 10/2011 of 14 January 2011 on plastic materials and articles intended to come into contact with food. Official Journal of the European Union, L12, 1-89. 1 1 2 3 Table 1 4 Details of the twelve plastic films. 5 6 7 8 9 10 11 12 13 14 15 16 17 Sample Code Type of material Thickness µm Origin Observations Internal side External side 1EC PE PET 153 Ecuador With printing ink 2EC PE PET 141 Ecuador With printing ink 3EC PP PP 46 Ecuador Without printing ink 4EC PE PET 146 Ecuador Without printing ink 5EC PE PA 72 Ecuador Without printing ink 6ES PE PP 70 Spain Without printing ink 7ES PE EVA 70 Spain With printing ink 8ES PP PET 35 Spain With printing ink 9EC PE PA 50 Ecuador With printing ink 10ES PE EVA 61 Spain With printing ink 11ES PE PET 54 Spain With printing ink 12ES PE PET 77 Spain With printing ink PE: polyethylene PET: Polyethylene terephthalate PA: Polyamide PP: Polypropylene EVA: Ethylene vinyl acetate 1 Table 2 Purge and Trap Instrument Conditions Purge Parameters: Valve Oven Temp 140 °C Transfer Line Temp 140 °C Sample mount Temp 90 °C Purge ready Temp 35 °C Standby Flow 0 mL/min Pre-Purge Time 0.50 min Pre-Purge Flow 40 mL/min Sample Preheat time 1.00 min Sample Temp 60 °C Purge Time 20.00 min Purge Flow 40 mL/min Condenser Ready Temp 40 °C Condenser Purge Temp 20 °C Dry Purge Time 0.50 min Desorb Parameters: Desorb Preheat Temp 245 °C Desorb Time 2.00 min Desorb Temp 250 °C Desorb Flow 400 mL/min Bake Parameters Bake Time 10.00 min Bake Temp 280 °C Bake Flow 200 mL/min Condenser Bake Temp 200 °C 2 1 1 Table 3 Compounds extracted by dynamic Headspace and subsequently identified by GC-MS in the plastic materials with their respective level of toxicity (TC) according Cramer rules Compound IUPAC NAME CAS RT (min) TC Samples 1EC 2EC 3EC 4EC 5EC 6ES 7ES 8ES 9EC 10ES 11ES 12ES Hexane Hexane 110-54-3 6,07 I X X Hexane, 2,5-dimethyl- 2,5-dimethylhexane 592-13-2 10.56 I X Hexane, 2,4-dimethyl- 2,4-dimethylhexane 589-43-5 10.67 I X X Pentane, 2,3,4-trimethyl- 2,3,4- trimethylpentane 565-75-3 11.42 I X X X X X X Pentane, 2,3,3-trimethyl- 2,3,3- trimethylpentane 560-21-4 11.65 I X X X X X X X X Heptane, 3-methyl- 3-methylheptane 589-81-1 12.20 I X X X Cyclopentane, 1-ethyl-2- methyl-, cis- 1-ethyl-2- methylcyclopentane 930-89-2 13.05 I X X X Octane Octane 111-65-9 13.21 I X X X X X X X X X X 3-Heptene, 3-methyl- 3-Methyl-3-heptene 7300-03-0 13.30 I X 2-Heptene, 3-methyl- (E)-3-methylhept-2- ene 3404-75-9 13.39 I X X X 2-Octene oct-2-ene 111-67-1 13.95 I X X Heptane, 2,4-dimethyl- 2,4- dimethylheptane 2213-23-2 14.08 I X X Heptane, 2,5-dimethyl- 2,5- dimethylheptane 2216-30-0 14.58 I X X Octane, 3-methyl- 3-methyloctane 2216-33-3 14.58 I X Hexanal Hexanal 66-25-1 14.92 X Heptane, 2,3-dimethyl- 2,3- dimethylheptane 3074-71-3 15.42 I X X X Heptane, 3,4-dimethyl- 3,4- dimethylheptane 922-28-1 15.54 I X Heptane, 2,4,6-trimethyl- 2,4,6- trimethylheptane 2613-61-8 16.03 I X Heptane, 2,2,4-trimethyl- 2,2,4- trimethylheptane 14720-74-2 16.17 I X X X X X X Octane, 2,2-dimethyl- 2,2-dimethyloctane 15869-87-1 16.46 I X X X X 2 Octane, 3,3-dimethyl- 3,3-dimethyloctane 4110-44-5 16.98 I X X X X X Nonane, 4-methyl- 4-methylnonane 17301-94-9 17.62 I X X X X Compound IUPAC NAME CAS RT (min) TC Samples 1EC 2EC 3EC 4EC 5EC 6ES 7ES 8ES 9EC 10ES 11ES 12ES Octane, 2,3-dimethyl- 2,3-dimethyloctane 7146-60-3 17.70 I X Octane, 4-ethyl- 4-ethyloctane 15869-86-0 19.01 I X X X Nonane, 3-methyl- 3-methylnonane3- methylundecane 5911-04- 61002-43-3 19.52 I X Decane Decane 124-18-5 20.50 I X X X X X X X X X X X Octanal <n-> Octanal 124-13-0 22.24 I X Undecane Undecane 1120-21-4 23.82 I X X X X X X X X X 1-Tetradecanol tetradecan-1-ol 112-72-1 24.58 I X X X X Nonanal Nonanal 124-19-6 25.53 I X 1- Dodecene dodec-1-ene 112-41-4 26.80 I X X Dodecane Dodecane 112-40-3 26.89 I X X X X X X X X X X X X Benzene, 1,3-Bis(1,1- Dimethylethyl)- 1,3-ditert- butylbenzene 1014-60-4 29.32 I X X X Tridecane Tridecane 629-50-5 29.80 I X X X X X X X Tetradecane Tetradecane 629-59-4 32.50 I X X X X X X X Caprolactam Azepan-2-one 105-60-2 32.87 III X Octadecane Octadecane 593-45-3 35.06 I X x x x x X Butylated hydroxytoluene 2,6-ditert-butyl-4- methylphenol 128-37-0 36.79 II X X X X Diethyl phthalate diethyl benzene- 1,2-dicarboxylate 84-66-2 40.10 I X 2TC: Cramer Toxicity 3 1 Table 4 Confirmed and tentative identified compounds extracted by liquid extraction and identified by GC-MS in the plastic materials with their respective level of toxicity (TC) according Cramer rules. Compound IUPAC Name CAS RT (min) Function TC SML Samples 1EC 2EC 3EC 4EC 5EC 6ES 7ES 8ES 9EC 10ES 11ES 12ES *Dodecane Dodecane 112-40-3 12.09 Alkane I NL X X X X X X X X X X X X Triethylene glicol 2-[2-(2- hydroxyethoxy)eth oxy]ethanol 112-27-6 12.5 Solvent I ** X *Caprolactam azepan-2-one 105-60-2 13.14 Monomer in polyamides III 15 X X X X *Tridecane Tridecane 629-50-5 13.91 alkane I NL X Tripropylene Glycol 1-[2-(2- hydroxypropoxy)pr opoxy]propan-2-ol 1638-16-0 14.30 Solvent (solubilize resins commonly used in printingink) internet III NL X *2,6-Toluene diisocyanate 1,3-diisocyanato-2- methylbenzene 91-08-7 14.75 Used in the production of flexible polyurethane foams III ND X *2,4-Toluene diisocyanate 2,4-diisocyanato-1- methylbenzene 584-84-9 14.85 Used in the production of flexible polyurethane foams III ND X X X *Tetradecane Tetradecane 629-59-4 15.61 Alkane I NL X X X X X 2,6-di-tert-butyl-1,4- benzoquinone 2,6-ditert- butylcyclohexa-2,5- diene-1,4 dione 719-22-2 16.65 Degradation product II NL X X X *Butylated Hydroxytoluene 2,6-ditert-butyl-4- methylphenol 128-37-0 17.29 Antioxidant II 3 X X X X 2,4-di-tert-butylphenol 2,4-ditert- butylphenol)- 96-76-4 17.32 Degradation product from antioxidants I NL X X X X X *Diethyl Phthalate diethyl benzene- 1,2-dicarboxylate 84-66-2 18.59 Solvent to hold color Plasticizer in the production of cellulose esters (cellulose acetate films) intended for I NL X X X X X X 2 contact with food Compound IUPAC Name CAS RT (min) Function TC SML Samples 1EC 2EC 3EC 4EC 5EC 6ES 7ES 8ES 9EC 10ES 11ES 12ES *Hexadecane Hexadecane 544-76-3 18.71 Alkane I NL X X X X X X X X *Benzophenone diphenylmethanone 119-61-9 19.25 Photoinitiator UV- inks III NL0,6 X X Benzenesulfonamide, N-ethyl-2-methyl- N-ethyl-2- methylbenzenesulf onamide 1077-56-1 19.61 Component of ink formulations III NL X X *Heptadecane Heptadecane 629-78-7 20.15 Alkane I NL X X X Benzenesulfonamide, N-ethyl-4-methyl N-ethyl-4- methylbenzenesulf onamide 80-39-7 20.47 Component of ink formulations III NL X X *Octadecane Octadecane 493-45-3 21.52 Alkane I NL X X X X X X Isopropyl myristate propan-2-yl tetradecanoate 110-27-0 21.82 Plasticizer, lubricant I NL X *Diisobutyl phthalate bis(2-methylpropyl) benzene-1,2- dicarboxylate 84-69-5 22.31 Plasticizer for PVC and other polymersPresent in printing inks I NL X X X X X 1-Octadecanol octadecan-1-ol 112-92-5 22.35 Ink solvent, plasticizer I NL X 1-Hexadecanol hexadecan-1-ol 36653-82-4 22.57 Adhesive in food packaging materials and coatings. I ** X X X 7,9 Di-Tert-Butyl-1- Oxaspiro (4,5)Deca-6- 9-Diene-2,8-Dione 7,9-ditert-butyl-1- oxaspiro[4.5]deca- 6,9-diene-2,8- dione 82304-66-3 22.90 Degradation product from antioxidant III NL X X X X X X X X X X X Methyl palmitate Methyl hexadecanoate 112-39-0 23.11 Intermediate for resins and defoamer in foodcontact coatings. I NL X 3 Palmitic acid Hexadecanoic acid 57-10-3 23.59 Slip agent degradant I ** X X X X X X X X X *Eicosane Eicosane 112-95-8 24.04 Alkane I NL X X X Compound IUPAC Name CAS RT (min) Function TC SML Samples 1EC 2EC 3EC 4EC 5EC 6ES 7ES 8ES 9EC 10ES 11ES 12ES Octadecanal Octadecanal 638-66-4 24.31 - I NL X Heptadecanol Heptadecan-1-ol 1454-85-9 25.05 Used as Plasticizer and in the manufacture of wetting agents and detergents. I NL X X X Tributyl aconitate Tributyl (1E)-1- propene-1,2,3- tricarboxylate 7568-58-3 25.85 Plasticizer I NL X X X X X Octadecanoic Acid Octadecanoic Acid 57-11-4 25.99 PVC lubricant, Adhesive in food packaging. I ** X Hexadecanamide Hexadecanamide 629-54-9 26.17 Slip agent III NL X X X *Docosane Docosane 629-97-0 26.35 Alkane I NL X X X X X X *Tributyl acetylcitrate tributyl 2- acetyloxypropane- 1,2,3- tricarboxylate 77-90-7 26.94 Plasticizer for flexible packaging films I 60 X X X X X X X X Oleamide (Z)-octadec-9- enamide 301-02-0 28.09 Slip agent III ** X X X X X X Octadecanamide Octadecanamide 124-26-5 28.34 Slip agent, antiblock agent and release agent. III ** X X *Bis(2-ethylhexyl) adipate bis(2-ethylhexyl) hexanedioate 103-23-1 28.40 Plasticizer in the production of plastic materials. I 18 60 X X 1-Docosanol Docosan-1-ol 661-19-8 28.42 Used in synthetic fibres and lubricants, thermal papers and I NL X 4 Toners *Tetracosane Tetracosane 646-31-1 28.48 Alkane I X X Triphenyl phosphate Triphenyl phosphate 115-86-6 28.50 Pigment additive Plasticizer for cellulose acetate articles and plasticizer for coatings, lacquers and varnishes III NL X X Compound IUPAC Name CAS RT (min) Function TC SML Samples 1EC 2EC 3EC 4EC 5EC 6ES 7ES 8ES 9EC 10ES 11ES 12ES *Bis(2-ethylhexyl) phthalate bis(2-ethylhexyl) benzene-1,2- dicarboxylate 117-81-7 29.85 Principal plasticizer in the production of PVC and its copolymers. Substance used as additives or ingredients of additive preparations I 1,5 60 X X X X cis-11-Eicosenamide (Z)-icos-11- enamide 10436-08-5 30.17 Used in adhesives and components of coatings III ** X X X *Erucamide (Z)-docos-13- enamide 112-84-5 32.15 Slip agent III ** X X X X X X X X X X X X *Squalene 2,6,10,15,19,23- hexamethyltetraco sa-2,6,10,14,18,22- hexaene 111-02-4 32.39 Used as oxygenscavenging I NL X X X X X X Glycerol tricaprylate 2,3- di(octanoyloxy)pro pyl octanoate 538-23-8 33.45 Lubricant I NL X 5 *Confirmed compounds TC: Cramer Toxicity SML: Specific migration limit IUPAC:PUB CHEM NL: Non listed in Regulation 10/2011 ** Specific migration limit of 60 mg/kg (Substances for which no specific migration limit or other restrictions are provided in Annex I Regulation 10/2011) Table 5 Comparison of the amounts of the compounds in packaging materials from Ecuador and migration into tenax and isooctane Compound RT (min) Samples (mg/dm2) 1EC 2EC 3EC 4EC 5EC 9EC PP I T PP I T PP I T PP I T PP I T PP I T Caprolactam 13.12 0.073 nd 0.025 0.68 nd 0.010 0.37 nd 0.024 Tetradecane 15.61 0.015 0.00024 0.0033 0.013 0.0014 0.0030 0.0056 0.00049 0.00095 2.6-di-tert-butyl-1.4-benzoquinone 16.65 0.013 nd 0.00074 2.4-di-tert-butylphenol 17.32 0.010 0.00057 0.00092 0.024 0.0016 0.00074 Diethyl Phthalate 18.56 0.023 0.0039 0.0025 0.0049 0.0012 0.0021 0.42 0.0014 0.0017 Hexadecane 18.71 0.0055 0.0015 0.0033 0.012 0.00055 0.0015 Benzenesulfonamide. N-ethyl-2-methyl- 19.55 0.049 0.0057 0.017 0.15 0.018 0.017 Heptadecane 20.15 0.0043 0.00015 0.0023 Benzenesulfonamide. N-ethyl-4-methyl 20.42 0.023 0.0021 0.0085 0.18 0.0054 0.0074 Octadecane 21.52 0.032 0.00049 0.0078 0.031 0.0015 0.0070 0.014 0.00062 0.0029 Isopropyl myristate 21.82 0.068 0.0041 0.017 Diisobutyl phthalate 22.31 0.0017 0.0014 0.00039 0.0030 0.0017 0.00038 1-Hexadecanol 22.57 0.030 nd 0.0084 7.9 Di-Tert-Butyl-1-Oxaspiro (4.5)Deca-6- 9-Diene-2.8-Dione 22.91 0.023 0.0016 0.0041 0.19 0.013 0.046 0.018 0.0024 0.0039 0.012 0.00075 0.0014 0.069 0.0029 0.0050 Octadecanal 24.31 0.017 0.011 0.0043 Tributyl aconitate 25.85 0.046 0.0096 0.025 Hexadecanamide 26.17 0.031 0.00093 0.0029 0.29 0.015 0.045 Docosane 26.35 0.019 0.0016 0.010 0.024 0.00085 0.013 0.0072 0.0010 0.0047 Tributyl acetyl citrate 26.94 0.088 0.046 0.010 2.0 0.17 0.27 Oleamide 28.09 0.30 0.016 0.051 Octadecanamide 28.34 0.22 0.0096 0.094 Triphenyl phosphate 28.50 0.12 0.039 0.044 0.37 0.062 0.15 Bis(2-ethylhexyl) phthalate 29.85 0.010 0.0011 0.0012 1.8 0.37 0.34 0.029 0.014 0.015 Erucamide 32.13 0.16 0.011 0.049 0.28 0.0011 0.024 0.52 0.085 0.13 1.3 0.14 0.19 0.55 0.026 0.063 0.81 0.088 0.18