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

Transcription

1 EFSA Journal 2012;10(7):2785 SCIENTIFIC PININ Scientific pinion on the safety and efficacy of benzyl alcohols, aldehydes, acids, esters and acetals (chemical group 23) when used as flavourings for all animal species 1 EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2, 3 European Food Safety Authority (EFSA), Parma, Italy ABSTRACT Chemical group 23 consists of benzyl alcohols/aldehydes/acids/esters/acetals, of which 36 are currently authorised for use as flavours in food. The use of the following is safe for all animal species: benzyl alcohol, benzoic acid and benzylacetate at the maximum of 125 mg/kg complete feed; benzylaldehyde, vanillin, gallic acid and benzylpropionate at the maximum of 25 mg/kg complete feed; 4-isopropylbenzyl alcohol, veratraldehyde, 4-isopropylbenzylaldehyde, p-tolualdehyde, benzylbutyrate, benzylformate, benzylisobutyrate, benzylisovalerate, benzylphenylacetate, methylbenzoate, ethylbenzoate, ethylsalicylate, methylsalicylate, isopentylbenzoate and isobutylbenzoate at the maximum of 5 mg/kg complete feed; p-methoxybenzaldehyde, p- anisylalcohol, p-anisylacetate, salicylaldehyde, 2-methoxybenzaldehyde, isopentylsalicylate, hexylsalicylate, isobutylsalicylate, benzylsalicylate and pentylsalicylate at the maximum of 1 mg/kg complete feed; benzylhexanoate, benzylcinnamate and benzylbenzoate at a maximum calculated use levels of 1.5 mg/kg complete feed for cattle, salmonids and non food-producing animals and 1.0 mg/kg complete feed for pigs and poultry; for piperonal the corresponding figures are 0.5 and 0.3 mg/kg complete feed. No safety concerns arise for the consumer following the use of benzoates, benzyl derivatives and salicylates up to the highest safe level in feedingstuffs for all animal species. Gallic acid, vanillin, veratraldehyde, p-anisylalcohol and p-anisylacetate are not expected to generate residues of safety concern when used in feedingstuffs for mammals and gallic acid in feedingstuffs for poultry. The lack of data for the other compounds in poultry and fish precludes an assessment of consumer exposure from these sources. For piperonal, no conclusion can be drawn on the safety for the consumer on the use of this substance in animal nutrition. All compounds should be considered as irritants to skin, eyes and respiratory tract, skin sensitisers and harmful if swallowed. The compounds do not pose a risk to the environment when used at concentrations considered safe for the target species. Since all compounds are used in food as flavourings, no further demonstration of efficacy is necessary. European Food Safety Authority, n request from the European Commission, Question No EFSA-Q , adopted by the FEEDAP Panel on 13 June 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, the late 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 Acknowledgement: 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 pinion on the safety and efficacy of benzyl alcohols, aldehydes, acids, esters and acetals (chemical group 23) when used as flavourings for all animal species. EFSA Journal 2012;10(7):2785. [30 pp.] doi: /j.efsa Available online: European Food Safety Authority, 2012

2 KEY WRDS Sensory additives, flavourings, phenyl ethyl alcohols, phenyl acetic acids, related esters, phenoxyacetic acids and related esters, chemical group 23, benzyl alcohol, 4-isopropylbenzyl alcohol, p-anisyl alcohol, benzaldehyde, 4-methoxy benzaldehyde, piperonal, veratraldehyde, vanillin, 4-isopropylbenzaldehyde, p- tolualdehyde, salicylaldehyde, 2-methoxy benzaldehyde, benzoic acid, gallic acid, benzyl acetete, p-anisyl acetate, benzyl butyrate, benzyl formate, benzyl propionate, benzyl hexanoate, benzyl isobutyrate, benzyl isovalerate, hexyl salicylate, benzyl phenylacetate, methyl benzoate, ethyl benzoate, benzyl benzoate, benzyl cinnamate, ethyl salicylate, methyl salicylate, isobutyl salicylate, isopentyl salicylate, benzyl salicylate, isopentyl benzoate, isobutyl benzoate, pentyl salicylate EFSA Journal 2012;10(7):2785 2

3 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 the safety and efficacy of 36 compounds (benzyl alcohols, aldehydes, acids, esters and acetals belonging to chemical group 23) when used as flavourings for all animal species. All are currently authorised for use as flavours in food and all have been detected in plant materials, in fruits or in processed foods, however the reports of their distribution vary greatly. The FEEDAP Panel concludes that the use of the following compounds is safe for all animal species: benzyl alcohol, benzoic acid and benzyl acetate at the maximum level of 125 mg/kg complete feed; benzylaldehyde, vanillin, gallic acid and benzyl propionate at the maximum level of 25 mg/kg complete feed; 4-isopropylbenzyl alcohol, veratraldehyde, 4-isopropylbenzyl aldehyde, p- tolualdehyde, benzyl butyrate, benzyl formate, benzyl isobutyrate, benzyl isovalerate, benzyl phenylacetate, methyl benzoate, ethyl benzoate, ethyl salicylate, methyl salicylate, isopentyl benzoate and isobutyl benzoate at the maximum level of 5 mg/kg complete feed; p-methoxybenzaldehyde, p- anisyl alcohol, p-anisyl acetate, salicylaldehyde, 2-methoxybenzaldehyde, isopentyl salicylate, hexyl salicylate, isobutyl salicylate, benzyl salicylate and pentyl salicylate at the maximum level of 1 mg/kg complete feed; benzyl hexanoate, benzyl cinnamate and benzyl benzoate at a maximum calculated use levels of 1.5 mg/kg complete feed for cattle, salmonids and non food-producing animals and 1.0 mg/kg complete feed for pigs and poultry; and piperonal at a maximum calculated use levels of 0.5 mg/kg complete feed for cattle, salmonids and non food-producing animals and 0.3 mg/kg complete feed for pigs and poultry. No safety concern would arise for the consumer from the use of benzoates, benzyl derivatives and salicylates up to the highest safe level in feedingstuffs for all animal species. Gallic acid, vanillin, veratraldehyde, p-anisyl alcohol and p-anisyl acetate are not expected to generate residues of safety concern when used in feddingstuffs for mammals and gallic acid in feedingstuffs for poultry. The lack of data on metabolism of the other compounds in poultry and fish precludes an assessment of consumer exposure from these sources. For piperonal, no conclusion can be drawn on the safety for the consumer on the use of this substance in animal nutrition. The FEEDAP Panel considers it prudent to treat all compounds under assessment as irritants to skin, eyes and respiratory tract, skin sensitisers and harmful if swallowed. The compounds assessed are not expected to pose a risk to the environment when used at the concentrations considered safe for the target species. Since all 36 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. EFSA Journal 2012;10(7):2785 3

4 TABLE F CNTENTS Abstract... 1 Summary... 3 Table of contents... 4 Background... 5 Terms of reference... 5 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) Safety Safety for the target species Conclusions on the safety for target species Safety for the consumer Benzoates and benzyl derivatives Salicylates ther CG 23 compounds Conclusions on the safety for the consumer Safety for the user Safety for the environment Efficacy Conclusions Documentation provided to EFSA References Appendices EFSA Journal 2012;10(7):2785 4

5 BACKGRUND 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 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 eighteen substances listed in Table 1 belonging to chemical group 23 benzyl alcohols, aldehydes, acids, esters and acetals, to be used as feed additives for all animal 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 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 1 July The additives are listed as food and feed flavourings in the register of Flavouring substances (CD 217/1999) 7 and in the European Union Register of Feed Additives, respectively. They have not been previously assessed by EFSA for this purpose. The 36 substances belonging to CG 23 have been previously assessed by JECFA (2001a, 2002b, 2002c) and EFSA (2005, 2008a, 2008b, 2009 and 2011) as food flavourings. TERMS F 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 animals, 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. J L 268, , p 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 ctober J L 84, , p. 1. EFSA Journal 2012;10(7):2785 5

6 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 Chemical defined flavourings from Chemical Group 23: 2-Methoxybenzaldehyde 4-Isopropylbenzaldehyde 4-Isopropylbenzyl alcohol 4-Methoxybenzaldehyde Benzaldehyde Benzoic acid Benzyl acetate Benzyl alcohol Benzyl benzoate Benzyl butyrate Benzyl cinnamate Benzyl formate Benzyl hexanoate Benzyl isobutyrate Benzyl isovalerate Benzyl phenylacetate Benzyl propionate Benzyl salicylate Ethyl benzoate Ethyl salicylate Gallic acid Hexyl salicylate Isobutyl benzoate Isobutyl salicylate Isopentyl benzoate Isopentyl salicylate Methyl benzoate Methyl salicylate p-anisyl acetate p-anisyl alcohol Pentyl salicylate Piperonal p-tolualdehyde Salicylaldehyde Vanillin Veratraldehyde - 2. Sensory additives b) flavouring compounds Composition, description 2-Methoxybenzaldehyde (CAS No ) 4-Isopropylbenzaldehyde (CAS No ) 4-Isopropylbenzyl alcohol (CAS No ) 4-Methoxybenzaldehyde (CAS No ) Chemical formula Description Purity criteria (if appropriate) C 8 H % C 10 H 12 95% C 10 H 14 97% C 8 H % Method of analysis (if appropriate) EFSA Journal 2012;10(7):2785 6

7 Benzaldehyde (CAS No ) C 7 H 6 98% Benzoic acid (CAS No ) C 7 H % Benzyl acetate (CAS No ) C 9 H % Benzyl alcohol (CAS No ) C 7 H 8 98% Benzyl benzoate (CAS No ) C 14 H % Benzyl butyrate (CAS No ) C 11 H % Benzyl cinnamate (CAS No ) 8 C 16 H % Benzyl formate (CAS No ) C 8 H % Benzyl hexanoate (CAS No ) C 13 H % Benzyl isobutyrate (CAS No ) C 11 H % Benzyl isovalerate (CAS No ) C 12 H % Benzyl phenylacetate (CAS No ) C 15 H % Benzyl propionate (CAS No ) C 10 H % Benzyl salicylate (CAS No ) C 14 H % Ethyl benzoate (CAS No ) C 9 H % Ethyl salicylate (CAS No ) C 9 H % Gallic acid (CAS No ) C 7 H % Hexyl salicylate (CAS No ) C 13 H % Isobutyl benzoate (CAS No ) C 11 H % Isobutyl salicylate (CAS No ) C 11 H % Isopentyl benzoate (CAS No ) C 12 H % Isopentyl salicylate (CAS No ) C 12 H % Methyl benzoate (CAS No ) C 8 H % Methyl salicylate (CAS No ) C 8 H % p-anisyl acetate (CAS No ) C 10 H % p-anisyl alcohol (CAS No ) C 8 H % Pentyl salicylate (CAS No ) C 12 H % Spectrometry (GC 8 Please note that benzyl cinnamate can be present in the trans- or cis-configuration. The natural occurrence of the substance is the trans-configuration, which most references refer to. EFSA Journal 2012;10(7):2785 7

8 Piperonal (CAS No ) p-tolualdehyde (CAS No ) Salicylaldehyde (CAS No ) Vanillin (CAS No ) Veratraldehyde (CAS No ) C 8 H % C 8 H 8 97% C 7 H % C 8 H % C 9 H % 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) Specific conditions or restrictions for use (if appropriate) Specific conditions or restrictions for handling (if appropriate) Post-market monitoring (if appropriate) Specific conditions for use in complementary feedingstuffs (if appropriate) Marker residue ther provisions and additional requirements for the labelling - All feedingstuffs and water for drinking, as part of a premixture only - - 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(7):2785 8

9 ASSESSMENT 1. Introduction The Chemical Group (CG) 23 for flavouring substances is defined in Commission Regulation (EC) No 1565/ as benzyl alcohols/aldehydes/acids/esters/acetals. The present application concerns 36 compounds, which can be assigned to this CG. The flavours included in this assessment have all been detected in plant materials, fruits or in processed foods; however, the reports of their distribution vary greatly. Some are widely distributed (e.g. benzyl alcohol, benzaldehyde, vanillin, salicylaldehyde, benzoic acid, benzyl acetate, methyl benzoate, ethyl salicylate, and methyl salicylate) while for others there appears only a single reference to their occurrence in food (e.g. benzyl phenylacetate and isobutyl salicylate). All compounds except 2-methoxybenzaldehyde, benzyl hexanoate, hexyl salicylate and pentyl salicylate have been assessed by JECFA (2001a, 2002b and 2002c) and were considered safe for use in food without limit. Acceptable Daily Intake (ADI) values were specified for benzaldehyde, piperonal, vanillin, and methyl salicylate, and a group ADI was set for benzyl alcohol, benzoic acid, benzyl acetate and benzyl benzoate. EFSA considered all the compounds under assessment for use as food flavourings in Flavouring Group Evaluations (FGE) 20, FGE 52 and FGE 54 and reached similar conclusions for the compounds previously considered by JECFA (EFSA, 2005, 2008a, 2008b, 2009 and 2011). Both JECFA and EFSA evaluated tolualdehyde as a mixture of the o-, m- and p-isomers, whereas the present application is for p-tolualdehyde only (FGE 37). All 36 compounds are currently listed in the European Union database of flavouring substances and as such authorised for use in food. A consortium of companies (FFAC) supplying flavours to the feed industry has requested authorisation for the use of the substances listed in Table 2 as additives to feed and water for drinking (category: sensory additives, 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 and their physicochemical characteristics are summarised in Table 2. 9 Commission Regulation (EC) No 1565/2000 of 18 July 2000 laying down the measures necessary for the adoption of an evaluation programme in application of Regulation (EC) No 2232/96 of the European Parliament and of the Council. J L 180, , p 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. J L 133, , p EFSA Journal 2012;10(7):2785 9

10 Benzyl alcohol (02.010) 4-Isopropylbenzyl alcohol (02.039) p-anisyl alcohol (02.128) H H H Benzaldehyde (05.013) 4-Methoxybenzaldehyde (05.015) Piperonal (05.016) Veratraldehyde (05.017) Vanillin (05.018) 4-Isopropylbenzaldehyde (05.022) H p-tolualdehyde (05.029) Salicylaldehyde (05.055) 2-Methoxybenzaldehyde (05.129) Benzoic acid (08.021) H Gallic acid (08.080) Benzyl acetate (09.014) H H H H p-anisyl acetate (09.019) H Benzyl butyrate (09.051) Benzyl formate (09.077) Benzyl propionate (09.132) Benzyl hexanoate (09.316) Benzyl isobutyrate (09.426) Benzyl isovalerate (09.458) Hexyl salicylate (09.581) Benzyl phenylacetate (09.705) H Methyl benzoate (09.725) Ethyl benzoate (09.726) Benzyl benzoate (09.727) Benzyl cinnamate (09.738) Ethyl salicylate (09.748) Methyl salicylate (09.749) H H Trans form shown EFSA Journal 2012;10(7):

11 Isobutyl salicylate (09.750) Isopentyl salicylate (09.751) Benzyl salicylate (09.752) H Isopentyl benzoate (09.755) H Isobutyl benzoate (09.757) H Pentyl salicylate (09.762) H Figure 1: Molecular structures and FLAVIS numbers of flavourings of CG 23 Table 2: Chemically defined flavourings of CG 23 under application EU Register name CAS No. Flavis No. Molecular formula Molecular weight Physical status Benzyl alcohol C 7 H Liquid Isopropylbenzyl alcohol C 10 H ily liquid 2.53 p-anisyl alcohol C 8 H Liquid 1.1 Benzaldehyde C 7 H Liquid Methoxybenzaldehyde C 8 H Liquid 1.76 Piperonal C 8 H Solid 1.05 Veratraldehyde C 9 H Solid 1.22 Vanillin C 8 H Solid Isopropylbenzaldehyde C 10 H Liquid 3.17 p-tolualdehyde C 8 H Liquid 2.26 Salicylaldehyde C 7 H ily liquid Methoxybenzaldehyde C 8 H Solid 1.72 Benzoic acid C 7 H Solid 1.87 Gallic acid C 7 H Solid 0.7 Benzyl acetate C 9 H Liquid 1.96 p-anisyl acetate C 10 H Liquid 2.16 Benzyl butyrate C 11 H Liquid 3.06 Benzyl formate C 8 H Liquid 1.53 Benzyl propionate C 10 H Liquid 2.57 Benzyl hexanoate C 13 H Liquid 4.05 Benzyl isobutyrate C 11 H Liquid 2.50 Benzyl isovalerate C 12 H Liquid 3.26 Hexyl salicylate C 13 H Liquid - Benzyl phenylacetate C 15 H Liquid - Methyl benzoate C 8 H Liquid 2.12 Ethyl benzoate C 9 H ily liquid 2.64 Benzyl benzoate C 14 H ily liquid 3.97 Benzyl cinnamate C 16 H Solid 4.06 Ethyl salicylate C 9 H Liquid 2.95 Methyl salicylate C 8 H Liquid 2.55 Isobutyl salicylate C 11 H Liquid 4.00 Isopentyl salicylate C 12 H ily liquid 4.49 Benzyl salicylate C 14 H ily liquid 4.31 Isopentyl benzoate C 12 H ily liquid 4.15 Isobutyl benzoate C 11 H ily liquid 3.23 Pentyl salicylate C 12 H Liquid 4.36 Log K ow EFSA Journal 2012;10(7):

12 All 36 substances are produced by chemical synthesis. Typically several routes of synthesis are available and described in the dossier. 11 Data was provided on the batch to batch variation in five batches of each additive (with the exception of isopentyl benzoate and isobutyl benzoate where one and four batches were available, respectively). 12 The content of the active substance exceeded the JECFA specifications (Combined Compendium of Food Additives Specifications; JECFA, 2006) for all compounds (Table 3). Table 3: Identification of the substances and data on purity EU Register name JECFA specification % Assay % Average Range Benzyl alcohol > Isopropylbenzyl alcohol > p-anisyl alcohol > Benzaldehyde > Methoxybenzaldehyde > Piperonal > Veratraldehyde > Vanillin > Isopropylbenzaldehyde > p-tolualdehyde > Salicylaldehyde > Methoxybenzaldehyde > Benzoic acid > Gallic acid > Benzyl acetate > p-anisyl acetate > Benzyl butyrate > Benzyl formate > Benzyl propionate > Benzyl hexanoate > Benzyl isobutyrate > Benzyl isovalerate > Hexyl salicylate > Benzyl phenylacetate > Methyl benzoate > Ethyl benzoate > Benzyl benzoate > Benzyl cinnamate > Ethyl salicylate > Methyl salicylate > Isobutyl salicylate > Isopentyl salicylate > Benzyl salicylate > Isopentyl benzoate > (a) Isobutyl benzoate > (b) 99.7 Pentyl salicylate > (a) ne batch available; (b) Average of four batches 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. 11 Technical dossiers/section II. 12 Technical dossiers/section II/Annex 2.1 and Supplementary Information June EFSA Journal 2012;10(7):

13 2.2. Stability and homogeneity A minimum shelf-life for all compounds is 12 months with the majority stable for a longer period when stored in closed containers under recommended conditions (in a cool and dry place). This assessment is made on the basis of compliance with the original specification after storage. Although no data is required for the stability of volatile additives in premixes and feed, 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 administration Conditions of use The applicant proposes the use of the 36 additives in feed or water for drinking for all animal species without withdrawal. 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. EFSA Journal 2012;10(7):

14 Table 4: Proposed dose for feed use EU Register name Feed concentration (mg/kg) Normal High Benzyl alcohol Isopropylbenzyl alcohol 1 5 p-anisyl alcohol 1 5 Benzaldehyde Methoxybenzaldehyde 5 25 Piperonal 1 5 Veratraldehyde 1 5 Vanillin Isopropylbenzaldehyde 1 5 p-tolualdehyde 1 5 Salicylaldehyde Methoxybenzaldehyde 1 5 Benzoic acid Gallic acid Benzyl acetate p-anisyl acetate 1 5 Benzyl butyrate 1 5 Benzyl formate 1 5 Benzyl propionate 5 25 Benzyl hexanoate 1 5 Benzyl isobutyrate 1 5 Benzyl isovalerate 1 5 Hexyl salicylate 1 5 Benzyl phenylacetate 1 5 Methyl benzoate 1 5 Ethyl benzoate 1 5 Benzyl benzoate 1 5 Benzyl cinnamate 1 5 Ethyl salicylate 1 5 Methyl salicylate 5 25 Isobutyl salicylate 1 5 Isopentyl salicylate 1 5 Benzyl salicylate 5 25 Isopentyl benzoate 1 5 Isobutyl benzoate 1 5 Pentyl salicylate 1 5 No specific proposals are made for doses to be used in water for drinking. However the FEEDAP Panel notes that 24 out of the 36 compounds are described as insoluble in water, and 9 compounds are considered either slightly or very slightly soluble 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 Chemically Defined Flavourings Group 23 (CDG23 Benzyl alcohols, aldehydes, acids, esters and acetals) in animal feed. The Executive Summary of the EURL report can be found in Appendix A. EFSA Journal 2012;10(7):

15 3. Safety The assessment of safety is based on the high use level proposed by the applicant (125 mg/kg complete feed for benzyl alcohol, vanillin, benzoic acid, gallic acid and benzyl acetate, 25 mg/kg complete feed for benzaldehyde, 4-methoxybenzaldehyde, benzyl propionate, methyl salicylate and benzyl salicylate and 5 mg/kg complete feed for the other additives, see 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 data for human exposure in the EU (2008, 2009 and 2011) range between and µg/person per day. Table 5 summarises the result of the comparison with human exposure for representative target animals. The body weight of target animals is taken from the default values shown in Table 8 (Appendix B). EFSA Journal 2012;10(7):

16 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 075 )/day* Target animal exposure µg/mbw (kg 075 )/day Salmon Piglet Dairy cow Benzyl alcohol Isopropylbenzyl alcohol p-anisyl alcohol Benzaldehyde Methoxybenzaldehyde Piperonal Veratraldehyde Vanillin Isopropylbenzaldehyde p-tolualdehyde Salicylaldehyde Methoxybenzaldehyde Benzoic acid Gallic acid Benzyl acetate p-anisyl acetate Benzyl butyrate Benzyl formate Benzyl propionate Benzyl hexanoate Benzyl isobutyrate Benzyl isovalerate Hexyl salicylate Benzyl phenylacetate Methyl benzoate Ethyl benzoate Benzyl benzoate Benzyl cinnamate ** Ethyl salicylate Methyl salicylate Isobutyl salicylate Isopentyl salicylate Benzyl salicylate Isopentyl benzoate Isobutyl benzoate Pentyl salicylate * mbw = metabolic body weight (kg 0.75 ) for a 60 kg person = 21.6; ** exposure based on EU intake figures as reported by JECFA The data in Table 5 clearly indicate that the intake by the target animals usually greatly exceeds that of humans, resulting from use in food for all 36 compounds. As a consequence, safety for the target species at the feed concentration applied cannot be derived from the risk assessment for food use. For benzoic acid safety data on a range of target species are available and have been previously considered (EFSA, 2012a). The use levels proposed for the use of benzoic acid as feed flavouring are considerably lower than the maximum use levels proposed for other uses of benzoic acid and salts (e.g mg/kg complete piglet feed; Regulation No (EC) 226/2012). 13 Therefore the FEEDAP Panel considers the use of benzoic acid safe at the levels proposed by the applicant. The FEEDAP 13 Commission Implementing Regulation (EU) No 226/2012 of 15 March 2012 amending Regulation (EC) No 1730/2006 as regards the conditions of use of benzoic acid (holder of authorisation Emerald Kalama Chemical BV). J L , p. 6. EFSA Journal 2012;10(7):

17 Panel extends this conclusion to benzoates (four compounds), benzyl alcohol and benzyl esters (nine compounds), benzaldehyde and 4-isopropylbenzyl alcohol taking into consideration chemical structure, metabolism and proposed use levels. For details of metabolism, see section Because of their much higher lipophilicity (LogK ow > 3.5) and their potential differences in toxicokinetics, three benzoates/benzyl esters were not included in this consideration. For these three compounds, namely benzyl hexanoate, benzyl cinnamate and benzyl benzoate, the threshold of toxicological concern (TTC) approach was followed (see below). For the remaining compounds, the maximum feed concentration which can be considered as safe for the target animal can be derived from the lowest No bserved Adverse Effect Level (NAEL) when suitable data is available. Toxicological data could only be found for six out of the remaining 16 compounds (reviewed by EFSA, 2011). Table 6 shows the available NAELs that are considered to be relevant for this group of additives. For a further nine compounds where a specific NAEL was unavailable a group NAEL approach was taken on the basis of structural and metabolic similarity, see section The NAEL for 4-methoxy benzaldehyde was extrapolated to p-anisyl alcohol and p-anisyl acetate, the NAEL for vanillin to veratraldehyde, the NAEL for salicylaldehyde to 2- methoxy benzaldehyde, the NAEL for methyl salicylate to ethyl salicylate and the NAEL for isopentyl salicylate to hexyl salicylate, isobutyl salicylate, benzyl salicylate and pentyl salicylate. EFSA Journal 2012;10(7):

18 Table 6: NAELs (mg/kg bw per day) used to derive the estimated safe feed concentration for the target species Flavouring 4-Methoxy benzaldehyde (05.015) NAEL mg/kg bw/day 20 (apply a safety factor of 200) Reference ECD SIDS, 2009 Vanillin (05.018) 400 Mancebo et al., 2003 Salicylaldehyde (05.055) 10 (apply a safety factor of 200) ECD SIDS, 2011 Gallic acid (08.080) 119 Niho et al., 2001 Methyl (09.749) Isopentyl salicylate (09.751) salicylate 50 Webb and Hansen, Drake et al., 1975 Study description, substance tested, end points 42-day study (0, 20, 100, 500 mg/kg bw per day), rat, gavage; effects on body weight, decrease platelet count, hyperplasia of squamous epithelium (all effects were observed at 500 mg/kg bw per day in males and 100 and 500 mg/kg bw per day in females, respectively) 13-week study (0, 80, 240, 400 mg/kg bw per day), rat, gavage; no effects on body weight, food consumption, hematological and biochemical parameters, organ weight, hystopathology 49-day study (0, 2.5, 10, 40, 160 mg/kg bw per day), rat, gavage; effects on liver hystopathology observed in the high dose groups (40 and 160 mg/kg bw per day) 13-week study (0, 0.2, 0.6, 1.7, 5 %), rat, diet; centrilobular liver cell hypertrophy, reflected in a significant increase in liver weight, was observed in animals of both sexes from 1.7 % No effects at 0.6 % (corresponding to 119 and 128 mg/kg bw per day in male and female rats, respectively) 2-year study (0, 0.1, 0.5, 1.0, 2.0 % corresponding to 0, 50, 250, 500, 1000 ng/kg bw per day), rat, diet; effects on growth, organ weight, haematological parameters, gross pathology observed at high dose levels No effects at 0.1 % 13-week study (0, 0.005, 0.05, 0.5 %), rat, diet; reduced body weight at 0.5 % associated with reduced food intake. Increased relative kidney weight without any histopathological changes at 0.05 and 0.5 % in the diet. No effects at % (corresponding to 4.7 mg/kg bw per day) Applying a safety factor of 100 to chronic and sub-chronic studies and 200 to studies of shorter duration to the NAEL, the maximum safe intake and thus the maximum safe feed concentration was calculated for different target species following the EFSA Guidance for sensory additives (EFSA, 2012b). For the remaining compounds, benzyl hexanoate, benzyl cinnamate and benzyl benzoate (Cramer class I) and piperonal (Cramer class II), the TTC approach was followed (EFSA Guidance on sensory additives, 2012b). The intake and body weight of target animals is taken from the default values given in Appendix B. The results of the calculations are shown in Table 8 (Appendix B). EFSA Journal 2012;10(7):

19 Table 7: Maximum safe concentration in feed of flavourings for different target animals EU Register name Maximum safe concentration in feed of a flavouring (mg/kg) Veal Calves Salmonids Cattle for fattening Pigs for fattening Sows Dairy cows Turkeys for fattening Piglets Chicken for fattening Laying hens Cats Dogs p-methoxybenzaldehyde p-anisyl alcohol p-anisyl acetate Vanillin Veratraldehyde Salicylaldehyde Methoxybenzaldehyde Gallic acid Methyl salicylate Ethyl salicylate Isopentyl salicylate Hexyl salicylate Isobutyl salicylate Benzyl salicylate Pentyl salicylate Benzyl hexanoate Benzyl cinnamate Benzyl benzoate Piperonal wing to their inability to metabolise drugs and phenolic compounds by glucuronidation, cats are particularly sensitive to adverse effects of salicylates. Cats are deficient in glucuronyl transferase and have a prolonged excretion of acetyl salicylate (half-life, 37.5 h). Acetyl salicylate is known to cause toxicosis in cats at doses > 25 mg/kg bw, with possible lethal effects at higher doses (325 mg). The same metabolic defect in the glucuronide pathway is responsible for the intolerance of cats to benzoic acid and benzyl alcohol (NRC, 1986). However, at the levels considered safe these effects are considered unlikely to occur Conclusions on the safety for target species The values for feed concentration are derived from a comparison between the concentration calculated as safe and those recommended as maximum levels by the applicant. They do not exceed the maximum specified by the applicant. n the basis of these considerations the FEEDAP Panel concludes that the use of the following compounds is safe for all animal species: - benzyl alcohol, benzoic acid and benzyl acetate at the maximum level of 125 mg/kg complete feed; - benzylaldehyde, vanillin, gallic acid and benzyl propionate at the maximum level of 25 mg/kg complete feed; - 4-isopropylbenzyl alcohol, veratraldehyde, 4-isopropylbenzyl aldehyde, p-tolualdehyde, benzyl butyrate, benzyl formate, benzyl isobutyrate, benzyl isovalerate, benzyl phenylacetate, methyl benzoate, ethyl benzoate, ethyl salicylate, methyl salicylate, isopentyl benzoate and isobutyl benzoate at the maximum level of 5 mg/kg complete feed; EFSA Journal 2012;10(7):

20 - p-methoxybenzaldehyde, p-anisyl alcohol, p-anisyl acetate, salicylaldehyde, 2- methoxybenzaldehyde, isopentyl salicylate, hexyl salicylate, isobutyl salicylate, benzyl salicylate and pentyl salicylate at the maximum level of 1 mg/kg complete feed. The calculated safe use levels for benzyl hexanoate, benzyl cinnamate and benzyl benzoate are 1.5 mg/kg complete feed for cattle, salmonids and non food-producing animals and 1.0 mg/kg complete feed for pigs and poultry. The equivalent values for piperonal are 0.5 and 0.3 mg/kg complete feed, respectively. Safe concentrations should be appropriately reduced if used in water for drinking. The absence of a margin of safety would not allow the simultaneous administration of the compounds under consideration except benzoic acid and related compounds in feed and water for drinking. If more than one salicylic acid derivative is used in flavouring premixtures the maximum levels should be reduced proportionally Safety for the consumer The safety for the consumer of all compounds used as food flavours has already been assessed by the CEF Panel (EFSA, 2005, 2008a, 2008b, 2009 and 2011) and JECFA (2002). All 36 compounds are currently authorised as food additives without limitations. Since the CEF opinion of 2011, one relevant paper has been published. An in vitro study in human lymphocytes with salts of benzoic acid (Zengin et al., 2011) reported positive results in chromosomal aberration, sister chromatid exchange and micronucleus frequency. The FEEDAP Panel considers that the new study would not require modification of the CEF position. As the intake of all 36 compounds by target animals exceeds that of humans resulting from use in food by one to three orders of magnitude, the metabolic fate and potential transfer of significant amounts of residues in edible tissues and products has to be considered Benzoates and benzyl derivatives In its evaluation of the chemical group benzoates and benzyl derivatives, JECFA (2002a) recognized that aromatic esters are absorbed rapidly in the gut and hydrolysed by the catalytic activity of carboxylesterases, which are found predominantly in the liver. Benzoate esters are hydrolysed to benzoic acid and aliphatic or aromatic alcohols, whereas benzyl derivatives generate benzyl alcohol and aliphatic and aromatic acids. The aliphatic alcohols are metabolized in the fatty acid pathway and tricarboxylic acid cycle. The aromatic alcohols, as well as benzaldehyde are oxidized to the corresponding aromatic acids which are subsequently β-oxidized (cinnamic acid) to benzoic acid which is excreted in the urine as a glycine conjugate. This is exemplified by the metabolic fate of benzylacetate where the absorption, routes of metabolism and excretion were apparently unaffected by the size or number of doses administered; no evidence of a reduction or saturation of the metabolic capacity of the experimental animals was observed in the dose range tested (Abdo et al., 1985). When high doses of benzyl derivatives are given, formation of the glycine conjugate is limited; when glycine is depleted, free benzoic acid may sequester acetyl coenzyme A or be excreted unchanged or as the glucuronic acid conjugate. JECFA stated also that the aromatic ring substitution such as in the ring-alkylsubstituted benzaldehyde (p-tolualdehyde and 4-isopropylbenzaldehyde) is not expected to modify the principal pathway of metabolism, the oxidation of the alkyl side-chain representing a minor route. In its evaluation of the same family of food flavouring compounds, EFSA (2011) concluded that the hydroxy- and methoxysubstituted benzyl derivatives are expected to be rapidly absorbed, metabolized (-demethylation of EFSA Journal 2012;10(7):

21 2-methoxybenzaldehyde and 4-methoxybenzaldehyde; oxidation of the aldehyde to the corresponding acid; conjugation of the hydroxyl group and the carboxylic group) and excreted mainly in the urine. The FEEDAP Panel (EFSA, 2012a) reported the extended capacity of mammals to metabolise benzoic acid even at high dose levels (up to 500 mg/kg bw), and the absence of bioaccumulation. It also stated that carboxylic acids including benzoic and salicylic acid follow a similar fate in chickens, conjugation occurring with ornithine instead of glycine. Studies in fish are scarce, but it was shown that benzoic acid was rapidly and extensively absorbed after oral administration to channel catfish, renal excretion being the primary route of elimination as unchanged benzoic acid and as a benzoyltaurine conjugate (Plakas and James, 1990) Salicylates The metabolic fate of salicylates has recently been assessed (EFSA, 2011). In mammals, salicylic acid esters are hydrolysed in the gastro-intestinal tract and the liver, yielding the corresponding alcohols (benzyl alcohol and aliphatic alcohols) and salicylic acid. Benzoyl alcohol is oxidised to benzaldehyde and benzoic acid, whereas salicylic acid is further metabolised in the liver via conjugation with glycine and subsequent formation of salicyluric acid, but also glucuronide conjugation. Given the rapid and near complete excretion of salicylates and related compounds in the urine, one can conclude that absorbed salicylates and their metabolites are widely distributed via blood, with little retention in tissues. Salicylic acid follows a similar fate in chickens, conjugation occurring with ornithine instead of glycine. No data are available for the metabolism of salicyclic acid in fish ther CG 23 compounds The major metabolic pathway of piperonal in rat, after oral administration (150 mg/kg bw), involved the oxidation and conjugation of the side chain; 94 % was excreted in the urine in 24 hours as piperonylic acid (about 20 %) and its glycine conjugate. The cleavage of the methylenedioxyphenyl moiety and expiration of the methylene carbon as carbon dioxide was demonstrated to occur to a very limited extent (Klungsoyr and Scheline, 1984). A similar metabolic fate of piperonal was described in mice (Kamienski and Casida, 1970). Gallic acid metabolism has been studied in the rat, rabbit and human (Booth et al., 1959; Parke, 1968; Shahrzad and Bitsch, 1998; Yasuda et al., 2000; Hodgson, 2000). After oral administration of gallic acid, the major metabolite identified in urine was free and 3--sulphated 4--methylgallic acid resulting from the methylation of the corresponding phenolic function. 3--methyl- and 3,4-dimethylgallic acid were identified in the human only, whereas the -demethylation of gallic acid was shown to occur in the rat and rabbit as the result of decarboxylation by the intestinal microflora, giving rise to pyrogallol and subsequently free and glucuronidated 2--methylpyrogallol. In chickens fed gallic acid the major urinary metabolite was identified as 4--methyl gallic acid, the secondary metabolite being conjugated and unconjugated pyrogallol (Potter and Fuller, 1968). The anaerobic degradation of gallic acid by partial decarboxylation to pyrogallol, reductive dehydroxylation at the 4-position to phloroglucinol and resorcinol, then lactone formation of the remaining carbonyl, was reported to occur in the rumen of sheep (Murdiati et al., 1992). Vanillin given orally to rats (100 mg/kg bw) resulted in the urinary excretion of the major metabolites (vanillin 7 %, vanillyl alcohol 19 % and vanillic acid 47 %) within the first 24 h. Vanillyl alcohol was conjugated mainly as glucuronide and/or sulphate conjugates, vanillic acid being excreted unchanged and as its glycine conjugate. Metabolisation by the intestinal bacteria of the conjugates excreted via the bile generated toluene derivatives and decarboxylated products (Strand and Scheline, 1975). Most of a dose of 100 mg/kg bw of vanillin administered by intraperitoneal injection to rabbits was excreted in urine in 24 h; about 69 % was oxidized to vanillic acid and 10 % reduced to vanillyl alcohol, whereas about 10% was excreted as the glucuronic acid conjugate of vanillin (Sammons and Williams, EFSA Journal 2012;10(7):

22 1941). JECFA (1968) reported that in man and rat, liver homogenates readily convert vanillin to vanillic acid, but also that vanillic acid was excreted in the urine following oral administration of vanillin. Veratraldehyde given orally to rabbits (2000 mg/kg bw) resulted in the urinary excretion of 70 % of the administered dose in 24 h as veratric acid and its glucuronoconjugate (Sammons and Williams, 1946). Presumably, veratric acid enters the enterohepatic circulation where gut microflora decarboxylates it to yield catechol. Decarboxylation of veratraldehyde to catechol (orthohydroxyphenol) has been shown to occur after incubation with rat caecal extracts (Scheline, 1972). p-anisyl acetate is likely to be de-acetylated in the digestive tract and liver by esterases, giving rise to p-anisyl alcohol which is oxidized to p-anisaldehyde (see ). Anisaldehyde given orally to rabbits (2000 mg/kg bw) resulted in the urinary excretion of 75 % of the administered dose into 24 h as the glucuronic acid conjugate of para-methoxybenzoic acid (anisic acid) (Sammons and Williams, 1946). The incubation of anisaldehyde with rat caecal preparations revealed the presence of anisic acid and anisyl alcohol (Scheline, 1972) Conclusions on the safety for the consumer Benzoates, benzyl derivatives and salicylates are rapidly absorbed, distributed, metabolised and excreted. Mammals, birds and fish share a similar metabolic capacity to handle these compounds and produce hydrophilic metabolites with low affinity for tissues which are excreted efficiently in the urine. The similarity of the metabolic fate of these substances in target and laboratory animals ensures that the consumer is exposed to the same residues as the laboratory animals used in toxicological studies. The transfer of the parent compounds is limited and no residues of safety concern derived from these compounds are to be expected. Consequently, no safety concern would arise for the consumer from the use of these compounds up to the highest safe level in feeds. Gallic acid, vanillin, veratraldehyde, p-anisyl alcohol and p-anisyl acetate are rapidly metabolized in mammals (including humans) through metabolic pathways and kinetics comparable to those followed by benzoates, benzyl derivatives and salicylates. Consequently they are not expected to generate residues of safety concern. Similar pathways have been demonstrated for gallic acid in chicken. The lack of data on metabolism of the other compounds in poultry and fish precludes an assessment of consumer exposure from these sources. For piperonal, the commonality of metabolic pathways in the laboratory animals, target species and humans has not been demonstrated. Consequently no conclusion can be drawn on the safety for the consumer when this substance is used in animal nutrition Safety for the user No experimental data on the safety for the user was provided. Hazards for skin and eye contact and respiratory exposure are recognised for 24 out of the 36 compounds. Twenty-three compounds are classified as irritating to eyes and/or skin. p-anisyl alcohol, 4-isopropylbenzaldehyde and benzyl salicylate may cause skin sensitisation, and salicylaldehyde is considered harmful in contact with skin. Thirteeen compounds (p-anisyl alcohol, aldehyde and salicylate derivatives) are identified as irritating to the respiratory system, and benzyl alcohol is considered harmful by inhalation. For the remaining substances, no hazards are identified, probably because the substances have not yet been tested. The FEEDAP Panel considers it prudent to treat all compounds under assessment as irritants to skin, eyes and respiratory tract, skin sensitisers and harmful if swallowed. EFSA Journal 2012;10(7):