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Safety and efficacy of essential oil, oleoresin and tincture from Zingiber officinale Roscoe when used as sensory additives in feed for all animal species

2020; Wiley; Volume: 18; Issue: 6 Linguagem: Inglês

10.2903/j.efsa.2020.6147

1831-4732

Vasileios Bampidis, Giovanna Azimonti, Maria de Lourdes Bastos, Henrik Christensen, Mojca Fašmon Durjava, Maryline Kouba, Marta López‐Alonso, Secundino López Puente, Francesca Marcon, Baltasar Mayo, Alena Pechová, Mariana Petkova, Fernando Ramos, Yolanda Sanz, Roberto Edoardo Villa, Ruud Woutersen, Paul Brantom, Andrew Chesson, Johannes Westendorf, Lucilla Gregoretti, Paola Manini, Birgit Dusemund,

Pesticide Exposure and Toxicity

EFSA JournalVolume 18, Issue 6 e06147 Scientific OpinionOpen Access Safety and efficacy of essential oil, oleoresin and tincture from Zingiber officinale Roscoe when used as sensory additives in feed for all animal species EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP), Corresponding Author EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) feedap@efsa.europa.eu Correspondence: feedap@efsa.europa.euSearch for more papers by this authorVasileios Bampidis, Vasileios BampidisSearch for more papers by this authorGiovanna Azimonti, Giovanna AzimontiSearch for more papers by this authorMaria de Lourdes Bastos, Maria de Lourdes BastosSearch for more papers by this authorHenrik Christensen, Henrik ChristensenSearch for more papers by this authorMojca Kos Durjava, Mojca Kos DurjavaSearch for more papers by this authorMaryline Kouba, Maryline KoubaSearch for more papers by this authorMarta López-Alonso, Marta López-AlonsoSearch for more papers by this authorSecundino López Puente, Secundino López PuenteSearch for more papers by this authorFrancesca Marcon, Francesca MarconSearch for more papers by this authorBaltasar Mayo, Baltasar MayoSearch for more papers by this authorAlena Pechová, Alena PechováSearch for more papers by this authorMariana Petkova, Mariana PetkovaSearch for more papers by this authorFernando Ramos, Fernando RamosSearch for more papers by this authorYolanda Sanz, Yolanda SanzSearch for more papers by this authorRoberto Edoardo Villa, Roberto Edoardo VillaSearch for more papers by this authorRuud Woutersen, Ruud WoutersenSearch for more papers by this authorPaul Brantom, Paul BrantomSearch for more papers by this authorAndrew Chesson, Andrew ChessonSearch for more papers by this authorJohannes Westendorf, Johannes WestendorfSearch for more papers by this authorLucilla Gregoretti, Lucilla GregorettiSearch for more papers by this authorPaola Manini, Paola ManiniSearch for more papers by this authorBirgit Dusemund, Birgit DusemundSearch for more papers by this author EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP), Corresponding Author EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) feedap@efsa.europa.eu Correspondence: feedap@efsa.europa.euSearch for more papers by this authorVasileios Bampidis, Vasileios BampidisSearch for more papers by this authorGiovanna Azimonti, Giovanna AzimontiSearch for more papers by this authorMaria de Lourdes Bastos, Maria de Lourdes BastosSearch for more papers by this authorHenrik Christensen, Henrik ChristensenSearch for more papers by this authorMojca Kos Durjava, Mojca Kos DurjavaSearch for more papers by this authorMaryline Kouba, Maryline KoubaSearch for more papers by this authorMarta López-Alonso, Marta López-AlonsoSearch for more papers by this authorSecundino López Puente, Secundino López PuenteSearch for more papers by this authorFrancesca Marcon, Francesca MarconSearch for more papers by this authorBaltasar Mayo, Baltasar MayoSearch for more papers by this authorAlena Pechová, Alena PechováSearch for more papers by this authorMariana Petkova, Mariana PetkovaSearch for more papers by this authorFernando Ramos, Fernando RamosSearch for more papers by this authorYolanda Sanz, Yolanda SanzSearch for more papers by this authorRoberto Edoardo Villa, Roberto Edoardo VillaSearch for more papers by this authorRuud Woutersen, Ruud WoutersenSearch for more papers by this authorPaul Brantom, Paul BrantomSearch for more papers by this authorAndrew Chesson, Andrew ChessonSearch for more papers by this authorJohannes Westendorf, Johannes WestendorfSearch for more papers by this authorLucilla Gregoretti, Lucilla GregorettiSearch for more papers by this authorPaola Manini, Paola ManiniSearch for more papers by this authorBirgit Dusemund, Birgit DusemundSearch for more papers by this author First published: 05 June 2020 https://doi.org/10.2903/j.efsa.2020.6147 Requestor: European Commission Question number: EFSA-Q-2017-00533 Panel members: Giovanna Azimonti, Vasileios Bampidis, Maria de Lourdes Bastos, Henrik Christensen, Birgit Dusemund, Maryline Kouba, Mojca Kos Durjava, Marta López-Alonso, Secundino López Puente, Francesca Marcon, Baltasar Mayo, Alena Pechová, Mariana Petkova, Fernando Ramos, Yolanda Sanz, Roberto Villa and Ruud Woutersen. Acknowledgements: The EFSA FEEDAP Panel (EFSA Panel on Additives and Products or Substances used in Animal Feed) wishes to thank the following for the support provided to this scientific output: Montserrat Anguita, Jaume Galobart and Fabiola Pizzo. Legal notice: Relevant information or parts of this scientific output have been blackened in accordance with the confidentiality requests formulated by the applicant pending a decision thereon by the European Commission. The full output has been shared with the European Commission, EU Member States and the applicant. The blackening will be subject to review once the decision on the confidentiality requests is adopted by the European Commission. Adopted: 7 May 2020 AboutSectionsPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Abstract Following a request from the European Commission, the EFSAPanel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of essential oil, oleoresin and tincture from Zingiber officinale Roscoe when used as sensory additives in feed for all animal species. The FEEDAPPanel concludes that the additives under consideration are safe for the target species at the following use levels: (i) ginger essential oil up to the maximum proposed use level of 80 mg/kg for veal calves (milk replacer) and 20 mg/kg complete feed (or 20 mg/L water for drinking) for all other species; (ii) ginger oleoresin at the maximum proposed concentrations of 20 mg/kg complete feed for fish, sheep, goats and horses and of 1 mg/kg for pets. For the remaining species, the calculated maximum safe concentration of ginger oleoresin in feed is less than that proposed use level and ranges from 5 mg/kg complete feed for chickens for fattening to 21 mg/kg for veal calves; (iii) ginger tincture at the maximum proposed concentrations of 1.6 mL/kg complete feed for horses and 0.26 mL/kg for dogs. For poultry species, the calculated maximum safe dose ranges between 0.2 and 0.3 mg/L water for drinking. No concerns for consumers were identified following the use of the additives up to the highest safe level in animal nutrition. The additives should be considered as irritants to skin and eyes and the respiratory tract and as a skin sensitisers. The use of the additives in feed is not expected to pose a risk for the environment. Since ginger and its preparations are recognised to flavour food and their function in feed would be essentially the same as that in food, no further demonstration of efficacy is considered necessary. 1 Introduction 1.1 Background and Terms of Reference Regulation (EC) No 1831/20031 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 and in addition, Article 10(2) of that Regulation specifies that for existing products within the meaning of Article 10(1), an application shall be submitted in accordance with Article 7, within a maximum of 7 years after the entry into force of this Regulation. The European Commission received a request from the Feed Flavourings Authorisation Consortium European Economic Interest Grouping (FFAC EEIG)2 for authorisation/re-evaluation of nine preparations (namely turmeric oil, oleoresin, extract (sb) and tincture from Curcuma longa L., cardamom oil from Elettaria cardamomum (L.) Maton, ginger oil, oleoresin, tincture and extract from Zingiber officinale Roscoe) belonging to botanically defined group (BDG) 9 - Zingiberales, when used as feed additives for all animal species (category: sensory additives; functional group: flavourings). During the course of the assessment, this application was split and the present opinion covers only four out of the nine preparations under application: ginger oil, ginger oleoresin, ginger tincture and ginger extract from Z. officinale for all animal species. During the assessment, the applicant withdrew the application for ginger extract.3 This preparation is excluded from the present assessment. 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. The particulars and documents in support of the application were considered valid by EFSA as of 3 January 2011.4 According to Article 8 of Regulation (EC) No 1831/2003, 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. EFSA shall deliver an opinion on the safety for the target animals, consumer, user and the environment and on the efficacy of the products ginger oil, ginger oleoresin and ginger tincture from Z. officinale, when used under the proposed conditions of use (see Sections 3.2.1.3, 3.3.1.3 and 3.4.1.3). The remaining five preparations belonging to botanically defined group (BDG) 9 - Zingiberales under application are assessed in separate opinions. 1.2 Additional information The three preparations under assessment, namely ginger oil, ginger oleoresin and ginger tincture from Z. officinale, are currently authorised as feed additives and listed in the European Union Register of Feed Additives pursuant to Regulation (EC) No 1831/2003 (2b natural products – botanically defined). They have not been assessed as feed additives in the EU. There is no specific EU authorisation for any Z. officinale preparation when used to provide flavour in food. However, according to Regulation (EC) No 1334/20085, flavourings preparations produced from food or food ingredients with flavouring properties may be used without an evaluation and approval as long as 'they do not, on the basis of the scientific evidence available, pose a safety risk to the health of the consumer, and their use does not mislead the consumer'. The European Medicines Agency (EMA, 2012) assessed Z. officinale, rhizome, as herbal medicinal product in the form of powdered herbal preparation. The European Food Safety Authority (EFSA NDA Panel, 2010) assessed the validity of the health claim on maintaining mobility of joints. 'Ginger' (Zingiberis rhizoma) is described in a monograph of the European Pharmacopoeia 10 (European Pharmacopoeia, 2019). It is defined as the dried, whole or cut rhizome of Z. officinale, with the cork removed, either completely or from the wide flat surfaces only, with a minimum content of essential oil of 15 mL/kg anhydrous drug. The preparations from Z. officinale are listed in the book of botanical flavourings of the Council of Europe with the number CoE 489/ Class N2- roots.6 2 Data and methodologies 2.1 Data The present assessment is based on data submitted by the applicant in the form of a technical dossier7 in support of the authorisation request for the use of ginger oil, ginger oleoresin and ginger tincture from Z. officinale as feed additives. The Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) used the data provided by the applicant together with data from other sources, such as previous risk assessments by EFSA or other expert bodies, peer-reviewed scientific papers, other scientific reports and experts' knowledge, to deliver the present output. Many of the components of the essential oil under assessment have been already evaluated by the FEEDAP Panel as chemically defined flavourings. The applicant submitted a written agreement to use the data submitted for the assessment of chemically defined flavourings (dossiers, publications and unpublished reports) for the risk assessment of preparations from Z. officinale.8 EFSA has verified the European Union Reference Laboratory (EURL) report as it relates to the methods used for the control of the phytochemical markers in the nine feed additives from botanically defined flavourings group 09 (BDG 09) – Zingiberales.9 The EURL delivered in 2018 an evaluation report related to the Botanically Defined Flavourings Group BDG 09 – Zingiberales. In this report, only analytical methods for cardamom oil were evaluated. On 25 February 2020, the EURL delivered an addendum to the above-mentioned report, in which the remaining feed additives included in this group were evaluated. In particular, regarding the feed additives subject of the present scientific opinion, the method of analysis for alpha-zingiberene, beta-sesquiphellandrene and ar-curcumene in ginger oil, and the methods of analysis for total gingerols and total shogaols in ginger oleoresin and ginger tincture were evaluated. For the ninth feed additive of this grouped application, namely ginger extract, the European Commission had accepted the request from the Applicant to withdraw the application for this feed additive. Therefore, the analytical methods for ginger extract were not evaluated in this addendum. The full report including the addendum is available on the EURL website: https://ec.europa.eu/jrc/en/eurl/feed-additives/evaluation-reports/fad-2010-0335. 2.2 Methodologies The approach followed by the FEEDAP Panel to assess the safety and the efficacy of ginger oil, ginger oleoresin and ginger tincture from Z. officinale is in line with the principles laid down in Regulation (EC) No 429/200810 and the relevant guidance documents: Guidance on safety assessment of botanicals and botanical preparations intended for use as ingredients in food supplements (EFSA SC, 2009), Compendium of botanicals that have been reported to contain toxic, addictive, psychotropic or other substances of concern (EFSA, 2012), Guidance for the preparation of dossiers for sensory additives (EFSA FEEDAP Panel, 2012a), Technical Guidance for assessing the safety of feed additives for the environment (EFSA, 2008), Guidance for the preparation of dossiers for additives already authorised for use in food (EFSA FEEDAP Panel, 2012b), Guidance on studies concerning the safety of use of the additive for users/workers (EFSA FEEDAP Panel, 2012c), Guidance on the safety of feed additives for the target species (EFSA FEEDAP Panel, 2017a), Guidance on the assessment of the safety of feed additives for the consumer (EFSA FEEDAP Panel, 2017b), Guidance document on harmonised methodologies for human health, animal health and ecological risk assessment of combined exposure to multiple chemicals (EFSA SC, 2019a), Statement on the genotoxicity assessment of chemical mixtures (EFSA SC, 2019b). 3 Assessment The additives under assessment are ginger oil, ginger oleoresin and ginger tincture from Zingiber officinale Roscoe and are intended for use as sensory additives (flavourings) in feed and water for drinking for all animal species. 3.1 Origin and extraction Ginger (Zingiber officinale Roscoe) is a perennial flowering plant which belongs to the Zingiberaceae family. The plant is probably native to Southeast Asia, but the wild type is no longer found and present-day ginger exists only as a cultigen. It is widely distributed and cultivated throughout Southeast Asia, and the Indian subcontinent and also commercially grown in Africa, China, Jamaica and other tropical parts of the world. The parts of the plant used for production of various culinary and medicinal preparations from the species are the fresh and/or dried rhizomes, which are designated by the name 'ginger' as the plant itself.11 The plant components present in the different preparations depend on the selectivity of the extraction process. The different extraction processes used for the additives which are the subject of this opinion, namely ginger essential oil, oleoresin and tincture, are described under their respective headings. 3.2 Ginger essential oil This application concerns the essential oil derived by steam distillation from the dried rhizomes of Z. officinale of Indian and Chinese origin. Briefly, steam is passed through the dried minced material from an external source or generated by boiling water below the material or by boiling water in which the material is immersed. The steam carries up the volatile constituents which are condensed. The essential oil is then separated from water by decantation. The amount of the essential oil present in rhizomes ranges between 1 and 5%, mainly containing α-zingiberene (70% in fresh and 20–30% in dried rhizomes).11 The molecular structures of the main components of the essential oil are shown in Figure 1. Figure 1Open in figure viewerPowerPoint Molecular formula of the main components of ginger essential oil 3.2.1 Characterisation of ginger essential oil The essential oil under assessment is a pale yellow to amber clear mobile liquid with a characteristic aroma of ginger. In 11 batches of the additive (five originating from China and six from India), the specific optical rotation at 25°C ranged between −38° and −32° (specification: −50° to −26°), the refractive index between 1.488 and 1.490 (specification: 1.484–1.498) and the density (25°C) between 874 and 881 kg/m3 (specification: 879–890).12 Ginger oil is identified with the single Chemical Abstracts Service (CAS) number 8007-08-7, European Inventory of Existing Chemical Substances (EINECS) number 283-634-2 and Flavour Extract Manufacturers Association (FEMA) 2522. The product specifications are based on the International Standard developed by the International Organisation for Standardization (ISO) 16928:2014 for essential oil of ginger [Zingiber officinale], which were adapted to reflect the concentrations of the main components of the essential oil, namely α-zingiberene (29–40%), β-sesquiphellandrene (8–14%), ar-curcumene (5–12%), α-farnesene (4–10%), camphene (2–10%) and β-bisabolene (2–9%). Analysis of 11 batches of the additive13 showed compliance with these specifications (Table 1). These six compounds account for about 70.8% on average (range 69.6–71.8%) of the product, expressed as area the area per cent (%) of the gas chromatographic (GC) profile. Table 1. Major constituents of the essential oil from the dried rhizomes of Zingiber officinale Roscoe as defined by the ISO standard: specifications and batch to batch variation based on the analysis of 11 batches. The content of each constituent is expressed as the area per cent of the corresponding chromatographic peak (% GC area), assuming the sum of chromatographic areas of all detected peaks as 100% Constituent Percentage of oil EU register name CAS No FLAVIS No Specification GC Area % Meana Range α-Zingiberene 495-60-3 – 29–40 36.78 33.82–39.20 β-Sesquiphellandrene 20307-83-9 – 8–14 10.25 9.22–11.23 ar-Curcumene 644-30-4 – 5–12 9.51 8.52–11.01 α-Farnesene 18794-84-8 01.014 4–10 6.84 4.45–9.32 Camphene 79-92-5 01.009 2–10 3.80 2.97–6.38 β-Bisabolene 495-61-4 01.028 2–9 3.65 2.47–4.24 Total 70.84 69.58–71.79 EU: European Union; CAS no.: Chemical Abstracts Service number; FLAVIS number: EU Flavour Information System numbers. a Mean calculated on 11 batches. The applicant provided the full characterisation of the 11 batches obtained by gas chromatography coupled with a flame ionisation detector (GC-FID) and mass spectrometry detector (GC-MS).13 In total, 103 constituents were detected, 91 of which were identified and accounted on average for 94.8% (92.8–95.4) of the product (as the GC area). Besides the six compounds indicated in the product specifications (accounting for 70.8% of the product), 13 other compounds were detected at levels > 0.5% and are listed in Table 2. These 19 compounds > 0.5% together account on average for 84.9% (83.2–86.0%) of the product. The remaining 72 compounds (ranging between 0.001% and 0.5% and accounting together for about 15% of the product) are listed in the footnote.14 Table 2. Constituents of the essential oil from the dried rhizomes of Zingiber officinale Roscoe accounting for > 0.5% of the composition (based on the analysis of 11 batches) not included in the specification. The content of each constituent is expressed as the area per cent of the corresponding chromatographic peak (% GC area), assuming the sum of chromatographic areas of all detected peaks as 100% Constituent Percentage of oil EU register name CAS No FLAVIS No Meana Range β-Phellandrene 555-10-2 01.055 3.04 2.47–4.22 Pin-2(3)-ene (α-Pinene) 80-56-8 01.004 1.44 1.17–2.38 1,8-Cineole 470-82-6 03.001 1.33 1.16–1.58 δ-Cadinene 29350-73-0 01.021 1.31 0.71–1.83 Germacra-1(10),4(14),5-triene 23986-74-5 01.042 1.17 0.91–1.46 D,L-Borneol 507-70-0 02.016 1.05 0.88–1.15 β-Elemene 33880-83-0 – 1.00 0.81–1.08 d-Limonene 5989-27-5 01.045 0.80 0.65–1.04 α-Terpineol 98-55-5 02.014 0.78 0.56–1.06 (-)-α-Elemol 639-99-6 02.149 0.57 0.49–0.65 trans-Nerolidol 7212-44-4 02.018 0.56 0.47–0.71 Farnesene (α and β) 18794-84-8 01.041 0.53 0.24–0.93 Alloaromadendrene 25246-27-9 – 0.52 0.31–0.66 Total 14.09 13.52–15.94 EU: European Union; CAS no.: Chemical Abstracts Service number; Flavis number: EU Flavour Information System numbers. a Mean calculated on 11 batches. Twenty-two peaks were detected but not identified, accounting in total for 3.60–6.34% (4.39% on average), with the single highest peak accounting on average for < 0.43% of the oil. The applicant performed a literature search regarding substances of concern and chemical composition of the plant species Z. officinale and its preparations.15 Perillene, 3-(4-methyl-3-pentenyl)-furan, CAS number 539-52-6, is a furan derivative that carries an alkenyl side chain in the 3-position. The occurrence of perillene has been reported in ginger rhizome essential oil. Analysis of the 11 batches showed an average content of 0.02% of perillene. 3.2.1.1 Impurities No information on the concentrations of undesirable compounds in the essential oil was given. The applicant makes reference to the 'periodic testing' of some representative flavourings premixtures for heavy metals (mercury, cadmium and lead), arsenic, fluoride, dioxins and dioxin-like polychlorinated biphenyls (PCBs), organo-chloride pesticides, organo-phosphorous pesticides, aflatoxin B1, B2, G1, G2 and ochratoxin A. Since ginger essential oil is produced by steam distillation, the likelihood of any measurable carry-over of heavy metals is low except for mercury (Tascone et al., 2014). 3.2.1.2 Shelf-life The typical shelf-life of ginger essential oil is stated to be at least 18 months, when stored in tightly closed containers under standard conditions (in a cool, dry place protected from light).16 No stability studies were performed for the essential oil. 3.2.1.3 Conditions of use Ginger essential oil is intended to be added to feed and water for drinking for all animal species without withdrawal period.17 The maximum proposed use level is 20 mg/kg complete feed for all species, except veal calves (milk replacer) and dairy cows, for which the maximum proposed use level is up to 150 mg/head and day which corresponds to 80 and 7.5 mg/kg (on dry matter basis), respectively. No specific use level has been proposed by the applicant for the use in water for drinking. 3.2.2 Safety The assessment of safety is based on the maximum use level proposed by the applicant. 3.2.2.1 Absorption, distribution, metabolism and excretion As no specific studies on absorption, distribution, metabolism and excretion (ADME) with the oil under assessment were provided, the ADME of the individual constituents is therefore considered. With the exception of camphene, which is a bicyclic monoterpenoid, the major components of the essential oil (α-zingiberene, ar-curcumene, β-sesquiphellandrene, β-bisabolene and α-farnesene) are structurally related sesquiterpenes (Figure 1). Other minor constituents are also aliphatic mono- or sesquiterpenes. Mono and sesquiterpenes are lipophilic compounds, which are expected to be rapidly absorbed from the gastro-intestinal tract and oxidised to polar oxygenated metabolites (by the cytochrome P450 enzymes, alcohol dehydrogenases and aldehyde dehydrogenases). The resulting hydroxylated metabolites may be excreted as glucuronide- and sulfate-conjugates or undergo further oxidation, yielding more polar metabolites that are also excreted in conjugated form in the urine and bile. Oxidation of the double bonds leads to epoxide intermediates which are rapidly detoxified either by hydrolysis to yield diols or by conjugation with glutathione. The enzymes involved in the biotransformation pathways of these compounds are present in all the target (food-producing and non-food producing) species (reviewed in EFSA FEEDAP Panel, 2016a). 3.2.2.2 Toxicological studies The applicant submitted a subchronic 90-day oral toxicity rat study performed with an essential oil from rhizome of Zingiber officinale (Jeena et al., 2011). Analysis shows that the essential oil tested is similar in composition and content to the essential oil under assessment (Table 3). Among the major components, the main differences are due to the different percentage of β-bisabolene, α-zingiberene and α-farnesene. However, these three compounds, accounting together for about 45% of the composition of the oils, are structurally related (see Figure 1) and have a similar toxicological profile. Table 3. Comparison of the test item used in the subchronic oral toxicity study (A) and the ginger essential oil under assessment (B) Constituent Essential oil A (%) Essential oil B (%) α-Zingiberene 31.08 36.8 (33.8–39.2) β-Sesquiphellandrene 14.02 10.2 (9.2–11.2) ar-Curcumene 15.35 9.5 (8.5–11.0) α-Farnesene – 6.8 (4.4–9.3) Camphene 5.14 3.8 (3.0–6.4) β-Bisabolene 13.81 3.7 (2.5–4.2) β-Phellandrene – 3.0 (2.5–4.2) Germacra-1(10),4(14),5-triene 2.40 1.2 (0.9–1.5) D,L-Borneol 1.14 1.0 (0.9–1.1) β-Elemene 2.14 1.0 (0.8–1.1) Alloaromadendrene 0.34 0.5 (0.3–07) 85.08 77.5 A total of 50 male and female Wistar rats (5 males and 5 females per group) were given 0 (control), 0 (vehicle control), 100, 250 or 500 mg essential oil A/kg body weight (bw) per day by oral gavage for 90 days. There were no deaths and no significant differences in growth between groups. The results of haematology, blood chemistry, gross pathology and histology showed no evidence of any treatment-related adverse effects. From this study, the highest dose tested (500 mg/kg bw per day) was identified by the authors as the no observed adverse effect level (NOAEL). The FEEDAP Panel agrees with the conclusions of the authors of the study. Ginger essential oil (A) was also tested for the induction of reverse mutations in Salmonella Typhimurium tester strains TA1535, TA98, TA100 and TA102 with or without metabolic activation, at five concentration levels up to 3 mg/plate. No indication of mutagenic activity was observed in any of the experimental conditions (Jeena et al., 2014). The FEEDAP Panel notes that for fully defined mixtures, the EFSA Scientific Committee (EFSA SC) recommends applying a component-based approach, i.e. assessing all components individually for their genotoxic potential (EFSA SC, 2019b). The ginger essential oil under assessment is well characterised (up to 94.8%) and all the components > 0.5% (except β-elemene and alloaromadendrene, for which no alerts were identified by QSAR analysis) have been assessed by EFSA for use in feed and/or food, and are currently authorised for food18 and feed19 uses. In these assessments, it was established that these compounds were not genotoxic. The remaining components were screened with OECD QSAR Toolbox and no alert were identified for in vitro mutagenicity (Ames test), for genotoxic and non-genotoxic carcinogenicity and for other endpoints. The Panel notes that at the maximum proposed use levels in feed, none of the components of the essential oil ranging in concentrations between 0.001% and 0.5%, including perillene, is considered of concern, as they are below the threshold of the corresponding Cramer Class, according to the threshold of toxicological concern (TTC) approach. 3.2.2.3 Safety for the target species Tolerance studies and/or toxicological studies made with the essential oil under application were not submitted. In the absence of these data, the approach to the safety assessment of the whole mixture can be based on read-across from a sufficiently similar mixture (EFSA SC, 2019a). The FEEDAP Panel considers the composition of the ginger essential oil tested in th