SCIENTIFIC OPINION. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2,3

Transcription

1 EFSA Journal 2012;10(2):2573 SCIENTIFIC OPINION Scientific Opinion on the safety and efficacy of phenol derivatives containing ring-alkyl, ring-alkoxy and side-chains with an oxygenated functional group (chemical group 25) when used as flavourings for all species 1 EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2,3 ABSTRACT European Food Safety Authority (EFSA), Parma, Italy Chemical Group 25 (CG 25) consist of phenol derivatives containing ring-alkyl, ring-alkoxy and side-chains with an oxygenated functional group, of which 16 are currently authorised for use as flavours in food. The flavouring compounds are calculated to be safe for all animal species at a maximum level of 5 mg/kg complete feed. This value supports the highest proposed use level for 13 of the 16 compounds with the margin of safety of approximately 1 to 5 for pigs and poultry, the most sensitive species. This safe level is substantially below the maximum proposed use level for all species for thymol and carvacrol (125 mg/kg complete feed) and for 3- methoxyphenol (25 mg/kg complete feed). The Panel concludes that the use of any of the 16 compounds included in CG 25 at the maximum proposed use level in animal feed would not raise safety concerns for consumers of animal products. No new data on the safety for the user was provided. However, potential hazards for skin and eye contact and respiratory exposure of various severities are recognised for all of the compounds included in CG 25. The Panel expects that at the use level safe for all target species (5 mg/kg feed), the 16 compounds included in CG 25 do not pose a risk for the environment. Since all 16 compounds include in CG 25 are used in food as flavourings and their function in feed is essentially the same as that in food, no further demonstration of efficacy was considered necessary. European Food Safety Authority, 2012 KEY WORDS Sensory Additives, Feed Flavourings, 2-methoxyphenol, thymol, 2-methoxy-4-methylphenol, 4-ethylguaiacol, 2-methoxy-4-vinylphenol, 4-ethylphenol, 3-methylphenol, 2-methylphenol, 4-methylphenol, carvacrol, 2,6- dimethoxyphenol, phenol, 2,6-dimethylphenol, 2-isopropylphenol, benzene-1,3-diol, 3,4-dimethylphenol 1 On request from the European Commission, Question No EFSA-Q , adopted on 01 February Revision /2012: minor editorial changes. 2 Panel members: Gabriele Aquilina, Georges Bories, Andrew Chesson, Pier Sandro Cocconcelli, Joop de Knecht, Noël Albert Dierick, Mikolaj Antoni Gralak, Jürgen Gropp, Ingrid Halle, Christer Hogstrand, Reinhard Kroker, Lubomir Leng, Secundino López Puente, Anne-Katrine Lundebye Haldorsen, Alberto Mantovani, Giovanna Martelli, Miklós Mézes, Derek Renshaw, Maria Saarela, Kristen Sejrsen and Johannes Westendorf. Correspondence: FEEDAP@efsa.europa.eu 3 The Panel wishes to thank the members of the Working Group on Feed Flavourings, including Paul Brantom, Joaquim Brufau, Gérard Pascal and Guido Rychen, for the preparatory work on this scientific opinion. Suggested citation: EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP); Scientific Opinion on the safety and efficacy of phenol derivatives containing ring-alkyl, ring-alkoxy and side-chains with an oxygenated functional group (chemical group 25) when used as flavourings for all species. EFSA Journal 2012;10(2):2573. [19 pp.] doi: /j.efsa Available online: European Food Safety Authority, 2012

2 SUMMARY Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on safety and efficacy of 16 phenol derivatives containing ring-alkyl, ring-alkoxy and side-chains with an oxygenated functional group (CG 25) when used as flavouring compounds in animal feed. All 16 compounds are currently authorised for use as flavours in food. The flavouring compounds are calculated to be safe for all animal species at a maximum level of 5 mg/kg complete feed. This value supports the highest proposed use level for 13 of the 16 compounds with the margin of safety of approximately 1 to 5 for pigs and poultry, the most sensitive species. This safe level is substantially below the maximum proposed use level for all species for thymol and carvacrol (125 mg/kg complete feed) and for 3-methoxyphenol (25 mg/kg complete feed). This concentration should be appropriately reduced when used in water for drinking. The Panel concludes that the use of any of the 16 compounds included in CG 25 at the maximum proposed use level in animal feed would not raise safety concerns for consumers of animal products. No new data on the safety for the user was provided. However potential hazards for skin and eye contact and respiratory exposure of various severity are recognised for all of the compounds included in CG 25. The Panel expects that at the use level safe for all target species (5 mg/kg feed), the 16 compounds included in CG 25 do not pose a risk for the environment. Since all 16 compounds include in CG 25 are used in food as flavourings and their function in feed is essentially the same as that in food, no further demonstration of efficacy was considered necessary. EFSA Journal 2012;10(2):2573 2

3 TABLE OF CONTENTS Abstract... 1 Summary... 2 Table of contents... 3 Background... 4 Terms of reference... 4 Assessment Introduction Characterisation Characterisation of the flavouring additives Stability and homogeneity Conditions of use Evaluation of the analytical methods by the European Union Reference Laboratory (EURL)11 3. Safety Safety for the target species Conclusions on the safety for target animals Safety for the consumer Safety for the user Safety for the environment Efficacy Documentation provided to EFSA References Appendix EFSA Journal 2012;10(2):2573 3

4 BACKGROUND Regulation (EC) No 1831/ establishes the rules governing the Community authorisation of additives for use in animal nutrition. In particular, Article 4(1) of that Regulation lays down that any person seeking authorisation for a feed additive or for a new use of a feed additive shall submit an application in accordance with Article 7; in addition, Article 10(2) of that Regulation also specifies that for existing products within the meaning of Article 10(1), an application shall be submitted in accordance with Article 7, at the latest one year before the expiry date of the authorisation given pursuant to Directive 70/524/EEC for additives with a limited authorisation period, and within a maximum of seven years after the entry into force of this Regulation for additives authorised without a time limit or pursuant to Directive 82/471/EEC. The European Commission received a request from the Feed Flavourings Authorisation Consortium European Economic Interest Grouping (FFAC EEIG) 5 for authorisation of the substances listed in Table 1 belonging to chemical group 25-phenol derivatives containing ring-alkyl, ring-alkoxy and side-chains with an oxygenated functional group, to be used as a feed additive for all species (category: sensory additives; functional group: flavourings) under the conditions mentioned in Table 1. According to Article 7(1) of Regulation (EC) No 1831/2003, the European Commission forwarded the application to the European Food Safety Authority (EFSA) as an application under Article 4(1) (authorisation of a feed additive or new use of a feed additive) and under Article 10(2) (re-evaluation of an authorised feed additive). EFSA received directly from the applicant the technical dossier in support of this application. 6 According to Article 8 of that Regulation, EFSA, after verifying the particulars and documents submitted by the applicant, shall undertake an assessment in order to determine whether the feed additive complies with the conditions laid down in Article 5. The particulars and documents in support of the application were considered valid by EFSA as of 02 February The additives are listed as food and feed flavourings in the register of Flavouring substances (CD 1999/217) 7 and in the Community Register of Feed Flavourings (Reg. (EC) No 1831/2003). They have not been previously assessed by EFSA for this purpose. The substances of the chemical group 25 have been previously assessed by JECFA (2001), and the EFSA Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Foods (AFC) (2006, 2008a, 2008b) as food flavourings. EMA assessed thymol and phenol when used as veterinary medical products (EMA, 1995 and 1996); no MRLs have been considered necessary to be set (Reg. (EC) No 37/2010). TERMS OF REFERENCE According to Article 8 of Regulation (EC) No 1831/2003, EFSA shall determine whether the feed additive complies with the conditions laid down in Article 5. EFSA shall deliver an opinion on the safety for the target animal(s), consumer, user and the environment and the efficacy of the active substances listed in Table 1, when used under the conditions described in Table 1. 4 Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22 September 2003 on additives for use in animal nutrition. OJ L 268, , p.29 5 Feed Flavourings Authorisation Consortium European Economic Interest Grouping (FFAC EEIG), Avenue Louise 130A, B-1050, Brussels, Belgium. 6 EFSA Dossier reference: FAD Commission Decision of 23 February 1999 adopting a register of flavouring substances used in or on foodstuffs drawn up in application of Regulation (EC) No 2232/96 of the European Parliament and of the Council of 28 October OJ L 84, , p. 1. EFSA Journal 2012;10(2):2573 4

5 Table 1: Description and conditions of use of the additive as proposed by the applicant Additive Registration number/ec No/No (if appropriate) Category(ies) of additive Functional group(s) of additive Chemically defined flavourings from Chemical Group 25 - Phenol derivatives: 2-Methoxyphenol Thymol 2-Methoxy-4-methylphenol 4-Ethylguaiacol 2-Methoxy-4-vinylphenol 4-Ethylphenol 3-Methylphenol 2-Methylphenol 4-Methylphenol Carvacrol 2,6-Dimethoxyphenol Phenol 2,6-Dimethylphenol 2-Isopropylphenol Benzene-1,3-diol 3,4-Dimethylphenol - 2. Sensory additives b) flavouring compounds Composition, description Chemical formula Description Purity criteria (if appropriate) 2-Methoxyphenol (CAS-Nr ) C 7 H 8 O 2 99 % Thymol (CAS-Nr ) C 10 H 14 O 98 % 2-Methoxy-4-methylphenol (CAS-Nr ) C 8 H 10 O 2 98 % 4-Ethylguaiacol (CAS-Nr ) C 9 H 12 O 2 98 % 2-Methoxy-4-vinylphenol (CAS-Nr ) C 9 H 10 O 2 96 % 4-Ethylphenol (CAS-Nr ) C 8 H 10 O 99 % 3-Methylphenol (CAS-Nr ) C 7 H 8 O 98 % 2-Methylphenol (CAS-Nr ) C 7 H 8 O 98 % 4-Methylphenol (CAS-Nr ) C 7 H 8 O 99 % Carvacrol (CAS-Nr ) C 10 H 14 O 98 % 2,6-Dimethoxyphenol (CAS-Nr ) C 8 H 10 O 3 98 % Phenol (CAS-Nr ) C 6 H 6 O 98 % 2,6-Dimethylphenol (CAS-Nr ) C 8 H 10 O 99 % 2-Isopropylphenol (CAS-Nr ) C 9 H 12 O 98 % Benzene-1,3-diol (CAS-Nr ) C 6 H 6 O 2 98 % 3,4-Dimethylphenol (CAS-Nr ) C 8 H 10 O 98 % Method of analysis (if appropriate) EFSA Journal 2012;10(2):2573 5

6 Trade name (if appropriate) - Name of the holder of authorisation (if appropriate) - Species or category of animal All species and categories Maximum Age Conditions of use Minimum content Maximum content mg or Units of activity or CFU/kg of complete feedingstuffs (select what applicable) Withdrawal period (if appropriate) Other provisions and additional requirements for the labelling Specific conditions or restrictions - for use (if appropriate) Specific conditions or restrictions All feedingstuffs and water for drinking for handling (if appropriate) Post-market monitoring - (if appropriate) Specific conditions for use in complementary feedingstuffs - (if appropriate) Marker residue Maximum Residue Limit (MRL) (if appropriate) Species or category of animal Target tissue(s) or food products Maximum content in tissues EFSA Journal 2012;10(2):2573 6

7 ASSESSMENT 1. Introduction The chemical group (CG) 25 for flavouring substances is defined in Commission Regulation (EC) No 1565/2000 as phenol derivatives containing ring-alkyl, ring-alkoxy, and side-chains with an oxygenated functional group. The present application concerns 16 separate compounds which meet this definition (see Table 2) and can be assigned to this CG. The majority of the 16 compounds are widely distributed in plants and plant derived products. However reports of the natural occurrence of the dimethoxyphenols, isopropylphenol and benzene -1,3-diol are more limited and appear restricted to manufacture products including dried fish. All 16 compounds have been assessed by JECFA (2001) and were considered safe for use in food. Subsequently EFSA, (2006, 2008) considered the same 16 compounds for use as food flavourings with similar conclusions. Phenol and thymol were assessed by EMA as veterinary drugs and no MRLs were considered necessary to be set (1995, 1996). Currently they are listed in the European Union database of flavouring substances and as such authorised for use in food. 8 The assessment of these substances as food flavourings excluded the risk of genotoxicity. A consortium of companies (FFAC) supplying flavours to the feed industry has requested authorisation for the use of the 16 compounds listed in Table 2 as additives in feed and water for drinking (category: sensory additives; functional group: flavouring compounds) for use in all animal species. Regulation (EC) No 429/ allows substances already approved for use in human food to be assessed with a more limited procedure than for other feed additives. However, the use of this procedure is always subject to the condition that food safety assessment is relevant to the use in feed. 2. Characterisation 2.1. Characterisation of the flavouring additives The molecular structures of the additives under application are shown in Figure 1, their physicochemical characteristics are summarised in Table Commission Regulation (EC) No 429/2008 of 25 April 2008 on detailed rules for the implementation of Regulation (EC) No 1831/2003 of the European Parliament and of the Council as regards the preparation and the presentation of applications and the assessment and the authorisation of feed additives. OJ L 133, , p. 1 EFSA Journal 2012;10(2):2573 7

8 2-Methoxyphenol (04.005) Thymol (04.006) 2-Methoxy-4-methylphenol (04.007) 4-Ethylguaiacol (04.008) 2-Methoxy-4-vinylphenol (04.009) 4-Ethylphenol (04.022) 3-Methylphenol (04.026) 2-Methylphenol (04.027) 4-Methylphenol (04.028) Carvacrol (04.031) 2,6-Dimethoxyphenol (04.036) Phenol (04.041) 2,6-Dimethylphenol (04.042) 2-Isopropylphenol (04.044) Benzene-1,3-diol (04.047) 3,4-Dimethylphenol (04.048) Figure 1: Molecular structures and FLAVIS numbers of Flavourings of CD 25 EFSA Journal 2012;10(2):2573 8

9 Table 2: Chemically defined flavourings from CG 25 under application Flavouring FLAVIS No CAS No Mol. Formula Mol. weight Phys. status pka Log K ow 2-Methoxyphenol C 7 H 8 O Solid Thymol C 10 H 14 O Solid Methoxy C 8 H 10 O Liquid methylphenol 4-Ethylguaiacol C 9 H 12 O Liquid Methoxy C 9 H 10 O Liquid - - vinylphenol 4-Ethylphenol C 8 H 10 O Solid Methylphenol C 7 H 8 O Liquid Methylphenol C 7 H 8 O Solid Methylphenol C 7 H 8 O Solid Carvacrol C 10 H 14 O Liquid ,6-Dimethoxyphenol C 8 H 10 O Solid Phenol C 6 H 6 O 94.1 Solid ,6-Dimethylphenol C 8 H 10 O Solid Isopropylphenol C 9 H 12 O Liquid Benzene-1,3-diol C 6 H 6 O Solid ,4-Dimethylphenol C 8 H 10 O Solid All the 16 substances are produced by chemical synthesis. Typical routes of chemical synthesis are available and described in details. 10 Data was provided on the batch to batch variation. 11 In each case, except for 3,4-dimethylphenol, the content of the active substance exceeded the JECFA specifications (JECFA, 2001) (Table 3) on which the assessment for food safety was made. 10 Technical dossier/section II 11 Technical dossier/section II/Annex 2.1 and Supplementary information March 2011 EFSA Journal 2012;10(2):2573 9

10 Table 3: Average of data on batch to batch variation and JECFA s specifications Flavouring FLAVIS No. JECFA specification % purity Average* Assay (%) Range 2-Methoxyphenol > Thymol > Methoxy-4-methylphenol > Ethylguaiacol > Methoxy-4-vinylphenol > Ethylphenol > Methylphenol > Methylphenol > Methylphenol > Carvacrol > ,6-Dimethoxyphenol > Phenol > ,6-Dimethylphenol > 99 Only one batch available Isopropylphenol > Benzene-1,3-diol > ,4-Dimethylphenol > 98 No data No data *Data based on 5 batches of the additives except otherwise specified Potential contaminants are considered as part of the product specification and are monitored as part of the HACCP procedure applied by all consortium members. The parameters considered include residual solvents, heavy metals and other undesirable substances Stability and homogeneity A shelf life of at least 24 months is given for all the chemicals under consideration except 3- methylphenol (60 months) and 2,6-dimethylphenol (36 months) when stored in closed containers under recommended conditions (in a cool, dry place, away from heat). This assessment is made on the basis of compliance with the original specification. Although no data is required for the stability of volatile additives in premixtures and feed, the use in water for drinking introduces other issues relating to product stability, such as degradation due to microbial activity. As no data on the short-term stability of the additive in water for drinking were provided, the FEEDAP Panel is not in the position to comment on this route of delivery. EFSA Journal 2012;10(2):

11 2.3. Conditions of use Phenol derivatives (CG 25) The applicant proposes the use of the 16 additives in feed or water for drinking for all animal species and without withdrawal time. In each case the applicant proposes a normal use level and a high use level of five times the normal level in feed as shown in Table 4. No specific proposals are made for doses to be used in water for drinking. However, the FEEDAP Panel notes that 12 compounds are described as insoluble or slightly soluble in water. Table 4: Conditions of use - Chemically defined flavourings from CG 25 under application Flavouring FLAVIS No. Feed concentrations (mg/kg) Normal High 2-Methoxyphenol Thymol Methoxy-4-methylphenol Ethylguaiacol Methoxy-4-vinylphenol Ethylphenol Methylphenol Methylphenol Methylphenol Carvacrol ,6-Dimethoxyphenol Phenol ,6-Dimethylphenol Isopropylphenol Benzene-1,3-diol ,4-Dimethylphenol Evaluation of the analytical methods by the European Union Reference Laboratory (EURL) EFSA has verified the EURL report as it relates to the methods used for the control of 2- methoxyphenol, thymol, 2-methoxy-4-methylphenol, 4-ethylguaiacol, 2-methoxy-4-vinylphenol, 4- ethylphenol, 3-methylphenol, 2-methylphenol, 4-methylphenol, carvacrol, 2,6-dimethoxyphenol, phenol, 2,6-dimethylphenol, 2-isopropylphenol, benzene-1,3-diol, 3,4-dimethylphenol in animal feed. The Executive Summary of the EURL report can be found in the Appendix. 3. Safety The assessment of safety is based on the high use level proposed by the applicant (5 mg/kg complete feed) for all compounds except 3-methylphenol (25 mg/kg complete feed) and thymol and carvacrol (125 mg/kg complete feed) (Table 4) Safety for the target species The first approach to the safety assessment for target species takes account of the applied use levels in animal feed relative to the maximum reported exposure of humans on the basis of the metabolic body weight. The intake and body weight of target animals is taken from the default values shown in Table 7. The data for human exposure for the 16 compounds range from to 11.6 µg/mbw(kg 0.75 ) /day. Table 5 summarises the result of the comparison with human exposure for selected target animals. EFSA Journal 2012;10(2):

12 Table 5: Comparison of exposure of humans and target animals to the flavourings under application Flavouring Use level in feed (mg/kg) Human exposure µg/mbw (kg 0.75 )/day* Target animal exposure µg/mbw (kg 0.75 )/day Salmon Piglet Dairy cow 2-Methoxyphenol Thymol Methoxy-4-methylphenol Ethylguaiacol Methoxy-4-vinylphenol Ethylphenol Methylphenol Methylphenol Methylphenol Carvacrol ,6-Dimethoxyphenol Phenol ,6-Dimethylphenol Isopropylphenol Benzene-1,3-diol ,4-Dimethylphenol * mbw: metabolic body weight (kg 0.75 ) for a 60 kg person = 21.6 The data in Table 5 confirms that the intake by the target animals given the maximum proposed dose substantially exceeds that of humans for all 16 compounds. As a consequence, safety for the target species at the feed concentration applied cannot be directly derived from the risk assessment for food use. The maximum feed concentration which can be considered as safe for the target animal can be derived from the lowest NOAEL available. Table 6 shows the available NOAELs that are considered to be relevant for this group of additives. Where no NOAEL could be identified for a particular compound a group NOAEL approach was taken on the basis of structural similarity. Where this was done, it is identified in Table 6. EFSA Journal 2012;10(2):

13 Table 6: Lowest NOAEL (mg/kg bw/day) for flavourings of CG 25 Flavouring NOAEL Reference Study description and end points 2-Methoxyphenol n.a. (*) Thymol 40 Matsuura, date unknown 12 2-Methoxy-4- methylphenol n.a. 4-Ethylguaiacol n.a. 2-Methoxy-4- vinylphenol n.a. 4-Ethylphenol n.a. 3-Methylphenol 50 Andersen, Methylphenol 50 Andersen, Methylphenol 50 Andersen, Carvacrol n.a. 2,6- Dimethoxyphenol n.a. 2,6-Dimethylphenol 100 Reproductive toxicity study in rats. Thickening of the forestomach, gross effects in adrenals and thymus Subchronic oral toxicity study with cresols on rats. Phenol 70 Ryan, Reduction in feed and water consumption Anonymous, Isopropylphenol n.a. Benzene-1,3-diol 36 3,4-Dimethylphenol (*) : No data available n.a NTP, 1992 On oral administration of 2,6-xylenol to rats by gavage for 4 weeks at a daily dose of 100 mg/kg body weight, an increase in liver weight was found without corresponding histopathological effects. Carcinogenicity study in mice (104 weeks): acute clinical signs (ataxia, tremors) Since individual NOAELs could be found only for seven out of 16 compounds the FEEDAP Panel took a pragmatic solution to identify a safe group level. This was based on the lowest NOAEL found for Benzene-1,3-diol (36 mg/kg bw). Applying a safety factor of 100 to this value, the maximum safe intake and thus the maximum safe feed concentration was derived for CG 25 and for different target species following the EFSA Guidance for sensory additives. The results of the calculations are shown in Table Included in the Draft Risk Assessment Report (DAR) for thymol used as pesticide Thymol_08_vol3_b6_tox_May_2011_public.pdf EFSA Journal 2012;10(2):

14 Table 7: Calculated maximum safe feed concentration of flavourings from CG 25 for different target animals (mg/kg) Target animal Default settings Maximum safe intake/feed concentration BW (*) (kg) FI (g/d) Intake (mg/d) mg/kg feed Salmonids Veal calves (milk replacer) Cattle for fattening Pigs for fattening Sows Dairy Cows Turkeys for fattening Piglets Chickens for fattening Laying hens Dog Cat (*) BW: body weight; FI: feed intake Conclusions on the safety for target animals The flavouring compounds included in CG 25 are calculated to be safe for all animal species at a maximum level of 5 mg/kg complete feed. A value of 5 mg/kg complete feed covers the high use level proposed by the applicant for 13 flavourings (2-methoxyphenol, 2-methoxy-4-methylphenol, 4-ethylguaiacol, 2-methoxy-4-vinylphenol, 4- ethylphenol, 2-methylphenol, 4-methylphenol, 2,6-dimethoxyphenol, phenol, 2,6-dimethylphenol, 2- isopropylphenol, benzene-1,3-diol and 3,4-dimethylphenol). The margin of safety for this use level varies between 1 and 5 for pigs and poultry, the most sensitive species. This concentration should be appropriately reduced when used in water for drinking. The simultaneous use in feed and water for drinking both at the highest safe concentration should be avoided. The high use levels proposed for 3-methylphenol (25 mg/kg) thymol and carvacrol (125 mg/kg) could not be confirmed as safe. The FEEDAP Panel also considered scenarios in which more than one member of CG 25 might be administered together at the highest proposed and highest safe use level, respectively. The Panel concludes that the safe limit shown in Table 7 should be respected regardless of the number of compounds of CG 25 simultaneously used to ensure the safety Safety for the consumer The safety for the consumer of all 16 compounds have been already assessed by the EFSA (FGE 22, 58 and 88) which endorsed the JECFA (JECFA, 2001) conclusions on the absence of safety concerns as food flavours. They are all authorised for use in food without limitation. JECFA (2001) and EFSA (2006, 2008) evaluations of the metabolic fate of phenol and alkyl-phenols concluded that these compounds are rapidly absorbed from the gastrointestinal tract, mainly conjugated with sulfate and glucuronic acid and excreted in the urine. If the capacity for conjugation is overwhelmed at high doses, alternative metabolic routes may be used including hydroxylation of the phenol ring (to quinol) and oxidation of the side-chains leading to the production of reactive metabolites (quinones, catechols, quinone methides). Alkoxy-phenols are O-demethylated and follow the same metabolic fate. An analysis of published data indicated that the metabolic pathways described for phenol are common to mammals (including the human), birds and fish (Kao et al, 1979; Capel et al., 1972; Layiwola and Linnecar, 1981; Kasokat et al., 1987). Tissue distribution in fish after repeated oral administration of alkylphenols with a longer alkyl chain (C4 to C7) than CG 25 members, i.e. with a higher lipophilicity and cumulative potential, indicated steady state EFSA Journal 2012;10(2):

15 concentrations after two days, very low residual values in the order: liver >> muscle and kidney, and a short biological half-life (Sundt and Baussant, 2003). These data suggest no accumulation of these phenolic compounds in tissues is expected. Consequently the Panel concludes that the use of the compounds of CG 25 in animal feed at the maximum concentrations defined in Table 7 would not raise safety concerns for consumers of animal products Safety for the user No new data on the safety for the user was provided. However potential hazards for skin and eye contact and respiratory exposure are recognised for all of the compounds included in CG 25. All carry risk phrases indicating effects of various severity ranging from skin and eye irritancy (R 36/38 in 9/16 cases) to clear corrosive effects (R 34 in 9/16 cases). Four compounds (4-Ethylguaiacol, 2-Methoxy-4- vinylphenol, 4-Ethylphenol, 2,6-Dimethoxyphenol) are identified as irritating to the respiratory system (R 37). In addition the potential hazards associated with phenol are well recognised and documented and work place exposure limits exist Safety for the environment All compounds in CG 25 to a varying extent occur naturally, primarily in plants. However, their natural distribution within European Union countries and the concentrations at which they normally exist vary greatly as do their environmental fates following emission. It is therefore difficult to a priori categorically state that the use of CG 25 compounds as flavourings in animal feeds will not raise their concentrations in the environment. Experimentally determined or structurally predicted degradation in soil and water is fast with a DT50 in soil ranging from about an hour (i.e., 4-methylphenol) to several weeks (e.g., 4-ethylphenol and carvacrol). Based on the predicted Koc value varying between 50 (benzene-1,3-diol) and 1050 (carvacrol), the CG 25 members are considered to be moderately mobile. Furthermore, the estimated bioconcentration factors for these compounds are all < 200 and, therefore, have a low potential for bioaccumulation. This confirmed with experimentally determined Bioconcentration Factor (BCF) values in fish for menthols which are in the range of < L/kg. Some of the CG 25 members are used as bio-pesticides because of their insecticide and/or insect repellent properties. Based on the available data (UK EPA, 2005; ENV.GOV.BC.CA, 2002) most CG 25 compounds can be considered to be acutely toxic to standard aquatic species (toxicity band 1 10 mg/l). Data on phenol, 2-methoxyphenol and benzene-1,3-diol, suggest that the chronic toxicity band is approximately 10 times lower. In view of the structural similarity and applying a safety factor of 10 to the lowest No Observed Effect Concentration (NOEC), the Predicted No Effect Concentration (PNEC) aquatic for the CG 25 members is not expected to be < 10 µg/l. The limited studies available on the toxicity to terrestrial species give Lethal (Effect) Concentration (L(E)C 50 ) values in range between 100 and > 500 mg/kg. Based on the Equilibrium Partitioning (EP) approach using a PNEC aquatic of 10 µg/l the PNEC soil will vary between µg/kg. Based on a dose of 5 mg/kg, which is considered safe for all target species, and assuming quantitative excretion as parent compound, the Predicted Environmental Concentration (PEC) soil will be close to the trigger value varying between µg/kg, assuming that manure will be plough into arable soil or be applied in several spreading events onto grassland. Using the screening method the PEC surface water will be < 10 µg/l based on the lowest Koc value. These PECs do not exclude a risk for the soil compartment. However, as mentioned in section 3.2, CG 25 compounds are also readily absorbed and biotransformed to conjugated metabolites by the target species and a small amount of the parent compounds are expected to be excreted. Even if CG 25 conjugated metabolites revert in the environment through microbial action, the regenerated parent compounds will also be subjected to 13 Directive 2009/161/EU establishing a third list of indicative occupational exposure limit values in implementation of Council Directive 98/24/EC and amending Commission Directive 2000/39/EC. OJ L , p. 87. EFSA Journal 2012;10(2):

16 biodegradation, which makes it unlikely that the maximum predicted concentrations will be reached. It is therefore expected that at a dose of 5 mg/kg the impact on the environment from the use of CG 25 compounds will be acceptably low. Consequently these compounds are not expected to pose a risk for the environment when used at a dose considered safe for the target species. 4. Efficacy Since these compounds are used in food as flavourings, and their function in feed is essentially the same as that in food no further demonstration of efficacy is necessary. CONCLUSIONS The flavourings of CG 25 are calculated to be safe for all animal species at a maximum level of 5 mg/kg complete feed. This value supports the highest proposed use level for 13 of the 16 compounds with the margin of safety of approximately 1 to 5 for pigs and poultry, the most sensitive species. The maximum safe level is substantially below the maximum proposed use level for all species for thymol and carvacrol (125 mg/kg complete feed) and for 3-methoxyphenol (25 mg/kg complete feed). This concentration should be appropriately reduced when used in water for drinking. The Panel concludes that the use of any of the 16 compounds included in CG 25 at the maximum proposed use level in animal feed would not raise safety concerns for consumers of animal products. No new data on the safety for the user was provided. However potential hazards for skin and eye contact and respiratory exposure of various severity are recognised for all of the compounds included in CG 25. The Panel expects that at the use level safe for all target species (5 mg/kg feed) the 16 compounds included in CG 25 do not pose a risk for the environment Since all 16 compounds include in CG 25 are used in food as flavourings, and their function in feed is essentially the same as that in food, no further demonstration of efficacy was considered necessary. DOCUMENTATION PROVIDED TO EFSA 1. Safety and efficacy of chemically defined flavourings from Chemical Group 25-Phenol derivatives containing ring-alkyl, ring-alkoxy and side-chains with an oxygenated functional group. December Submitted by the Feed Flavourings Authorisation Consortium European Economic Interest Grouping (FFAC EEIG). 2. Safety and efficacy of chemically defined flavourings from Chemical Group 25-Phenol derivatives containing ring-alkyl, ring-alkoxy and side-chains with an oxygenated functional group. Supplementary information. March Submitted by Feed Flavourings Authorisation Consortium European Economic Interest Grouping (FFAC EEIG). 3. Safety and efficacy of chemically defined flavourings from Chemical Group 25-Phenol derivatives containing ring-alkyl, ring-alkoxy and side-chains with an oxygenated functional group. Supplementary information. September Submitted by Feed Flavourings Authorisation Consortium European Economic Interest Grouping (FFAC EEIG). 4. Evaluation report of the Community Reference Laboratory for Feed Additives on the methods(s) of analysis for Chemically Defined Flavourings - Group 25 (CDG25, phenol derivatives containing ring-alkyl, ring-alkoxy, and side-chains with an oxygenated functional group) 5. Comments from Member States received through the ScienceNet. EFSA Journal 2012;10(2):

17 REFERENCES Andersen A, Final report on the safety assessment of sodium p-chloro-m-cresol, p-chloro-mcresol, chlorothymol, mixed cresols, m-cresol, o-cresol, p-cresol, isopropyl cresols, thymol, o- cymen-5-ol, and carvacrol. IJT. 25 Suppl 1: Anonymous, 1995.Toxikologische Bewertung. Heidelberg, Berufsgenossenschaft der chemischen Industrie Vol138: 19. Capel ID, French MR, Millburn P, The fate of foreign compounds in biological systems. Xenobiotica Volume: 2 Issue: 1 Pages: EFSA (European Food Safety Authority), Opinion of the Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food. Flavouring Group Evaluation 22: Ring-substituted phenolic substances from chemical groups 21 and 25. The EFSA Journal 393, EFSA (European Food Safety Authority), 2008a. Opinion of the Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food. Flavouring Group Evaluation 88, (FGE.88)[1] - Consideration of Phenol and Phenol Derivatives. The EFSA Journal, 857, s_en,3.pdf EFSA (European Food Safety Authority), 2008b. Opinion of the Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food. Flavouring Group Evaluation 58 (FGE.58) Consideration of phenol derivatives evaluated by JECFA (55th meeting) structurally related to ring substituted phenolic substances evaluated by EFSA in FGE.22 (2006) (Commission Regulation (EC) No 1565/2000 of 18 July 2000). The EFSA Journal, 711, EMA (European Agency for the Evaluation of Medicinal Products), Committee for Veterinary Medicinal Product. Thymol. EMEA/MRL/075/96-final. EMA (European Agency for the Evaluation of Medicinal Products), Committee for Veterinary Medicinal Product. Phenol. EMEA/MRL/040/95. ENV.GOV.BC.CA (Ministry of Water, Land and Air Protection. British Columbia. Canada), Ambient Working Water Quality Guidelines for Phenols. EPA UK (Environment Agency), 2005 Prioritisation of Alkylphenols for Environmental Risk Assessment. JECFA, Evaluation of certain food additives and contaminants. Fifty-fifth report of the Joint FAO/WHO Expert Committee on Food Additives. WHO Technical Report Series, no Geneva, 6-15 June Kao J, Bridges JW, Faulkner JK, Metabolism of C-14 Phenol by sheep, pig and rat. Xenobiotica 9 (3), Kasokat T, Nagel R, Urich K, The metabolism of phenol and substitued phenols in zebra fish. Xenobiotica 17 (10) Layiwola PJ, and Linnecar DFC, The biotransformation of Phenol (C-14) in some fresh-water fish. Xenobiotica 11 (3) NTP (National Toxicology Program), Toxicology and Carcinogenesis Studies of Resorcinol (CAS No ) in F344/N rats and B6C3F1 mice (gavage studies), NTP Technical Reports. Series No. 403, NTP, Research Triangle Park, NC, USA. EFSA Journal 2012;10(2):

18 Posternak JM Linder A and Vodoz CA, Summaries of toxicological data. Toxicological tests on flavoring matters. Food Chem Toxicol. 7, Ryan BM, Selby R, Gingell R, Waechter JM, Butala JH, Dimond SS, Dun BJ, House R and Morissey R, Two-generation reproduction study and immunotoxicity screen in rats dosed with phenol via the drinking water. Int J Toxicol 20 (3): Sundt RC and Baussant T, 2003Uptake, tissue distribution and elimination of C 4 C 7 alkylated phenols in cod. Dietary vs. waterborne exposure. Report AM-2003/ EFSA Journal 2012;10(2):

19 APPENDIX Executive Summary of the Evaluation Report of the European Union Reference Laboratory for Feed Additives on the Method(s) of Analysis for Chemically Defined Flavourings - Group 25 (CDG25, phenol derivatives containing ring-alkyl, ring-alkoxy, and side-chains with an oxygenated functional group) 14 The Chemically Defined Flavourings - Group 25 (CDG25, phenol derivatives containing ring-alkyl, ring-alkoxy, and side-chains with an oxygenated functional group), in this application comprises 16 substances, for which authorisation as feed additives is sought under the category "sensory additives", functional group 2(b) "flavouring compounds", according to the classification system of Annex I of Regulation (EC) No 1831/2003. In the current application submitted according to Article 4(1) and Article 10 (2) of Regulation (EC) No 1831/2003, the authorisation for all species is requested. The flavouring compounds of interest have a purity ranging from 96 % to 99 %. Mixtures of flavouring compounds are intended to be incorporated only into feedingstuffs or drinking water. The Applicant suggested no minimum or maximum levels for the different flavouring compounds in feedingstuffs. For the identification of volatile chemically defined flavouring compounds CDG25 in the feed additive, the Applicant submitted a qualitative multi-analyte gas-chromatography mass-spectrometry (GC-MS) method, using Retention Time Locking (RTL), which allows a close match of retention times on GC-MS. By making an adjustment to the inlet pressure, the retention times can be closely matched to those of a reference chromatogram. It is then possible to screen samples for the presence of target compounds using a mass spectral database of RTL spectra. The Applicant maintained two FLAVOR2 databases/libraries (for retention times and for MS spectra) containing data for more than 409 flavouring compounds. These libraries were provided to the EURL. The Applicant provided the typical chromatogram for the CDG25 of interest. In order to demonstrate the transferability of the proposed analytical method (relevant for the method verification), the Applicant prepared a model mixture of flavouring compounds on a solid carrier to be identified by two independent expert laboratories. This mixture contained twenty chemically defined flavourings belonging to twenty different chemical groups to represent the whole spectrum of compounds in use as feed flavourings with respect to their volatility and polarity. Both laboratories properly identified all the flavouring compounds in all the formulations. Since the substances of CDG 25 are within the volatility and polarity range of the model mixture tested, the Applicant concluded that the proposed analytical method is suitable to determine qualitatively the presence of the substances from CDG 25 in the mixture of flavouring compounds. Based on the satisfactory experimental evidence provided, the EURL recommends for official control for the qualitative identification in the feed additive of the individual (or mixture of) flavouring compounds of interest (listed in Table 1) the GC-MS-RTL (Agilent specific) method submitted by the Applicant. As no experimental data were provided by the Applicant for the identification of the active substance(s) in feedingstuffs and water, no methods could be evaluated. Therefore the EURL is unable to recommend a method for the official control to identify the active substance(s) of interest (cf. Table 1) in feedingstuffs or water. No further testing or validation is required. 14 The full report is available on the EURL website EFSA Journal 2012;10(2):