Safety and efficacy of a feed additive consisting of an essential oil derived from the flowering aerial parts of Salvia rosmarinus Spenn. (rosemary oil) for use in all animal species (FEFANA asbl)

TL;DR

This study evaluates the safety and effectiveness of rosemary oil as a feed additive for all animal species, finding it safe at specified concentrations.

Contribution

The paper provides a comprehensive safety assessment of rosemary oil as a feed additive across multiple animal species.

Findings

Rosemary oil is safe for use in feed at specified concentrations for various animal species.

Methyleugenol and estragole were detected in some batches but did not pose safety concerns at recommended levels.

The additive is considered an irritant and sensitiser, requiring precautions to minimize user exposure.

Abstract

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of an essential oil from the flowering aerial parts of Salvia rosmarinus Spenn. (rosemary oil) when used as a sensory additive in feed and in water for drinking for all animal species. Methyleugenol (< 0.039%) and estragole (< 0.027%) were detected in some batches of the additive. The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concluded that the use of rosemary oil is very unlikely to induce adverse effects in long‐living and reproductive animals and is of no safety concern for species for fattening at the following concentrations in complete feed: 18 mg/kg for chickens for fattening, 20.8 mg/kg for laying hens, turkeys for fattening, pigs for fattening, piglets, sows, veal calves (milk replacer), cattle for fattening, sheep/goats, horses, dairy cows, rabbits, salmonids and dogs, 500 mg/kg for ornamental fish and 14.1 mg/kg for cats. These conclusions were extrapolated to other physiologically related species. For any other species, the additive is very unlikely to induce adverse effects at 14.1 mg/kg complete feed. No concerns for consumers and the environment were identified following the use of the additive up to the maximum proposed use level in feed. Regarding user safety, the essential oil under assessment should be considered as an irritant to skin and eyes and as a dermal and respiratory sensitiser. When handling the additive, exposure of unprotected users to methyleugenol and estragole may occur. Therefore, to reduce the risk, the exposure of the users should be minimised. Since the leaves of S. rosmarinus are recognised to flavour food and their function in feed would be essentially the same as that in food, no further demonstration of efficacy was considered necessary.

Tables

TABLE 1. Constituents of rosemary oil as defined by specifications and batch‐to‐batch variation based on the analysis of 10 batches by gas chromatography with flame ionisation detector (GC‐FID). 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			% GC area
EU register name	CAS No	FLAVIS No	Specification 1	Mean	Range
1,8‐Cineole	470‐82‐6	03.001	14–62	34.0	17.4–48.8
Camphor 2	76‐22‐2	–	5–24	14.7	9.0–23.0
α‐Pinene	80‐56‐8	01.004	7–26	16.7	10.7–22.2
Camphene	79‐92‐5	01.009	< 15%	7.4	4.4–12.3
β‐Pinene	127‐91‐3	01.003	1–12	6.1	3.2–9.5

TABLE 2. Constituents of rosemary oil accounting for > 0.1% of the composition: Batch‐to‐batch variation based on the analysis of 13 batches by gas chromatography‐mass spectrometry (GC‐MS). 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			% GC area
EU register name	CAS No	FLAVIS No	Mean	Range
1,8‐Cineole	470‐82‐6	03.001	36.24	16.17–57.33
Camphor 1	76‐22‐2	–	15.29	11.12–23.13
α‐Pinene (pin‐2(3)‐ene)	80‐56‐8	01.004	14.67	8.60–19.74
β‐Pinene (pin‐2(10)‐ene)	127‐91‐3	01.003	5.31	2.87–9.26
Camphene	79‐92‐5	01.009	5.00	0.15–11.18
β‐Caryophyllene	87‐44‐5	01.007	3.13	1.31–5.61
d,l‐Borneol	507‐70‐0	02.016	3.06	0.13–4.60
d‐Limonene 2	5989‐27‐5	01.045	2.51	0.82–4.90
p‐Cymene (1‐isopropyl‐4‐methylbenzene)	99‐87‐6	01.002	2.27	1.36–2.95
α‐Terpineol	98‐55‐5	02.014	2.18	1.58–2.70
d,l‐Isoborneol	124‐76‐5	02.059	1.71	1.67–1.75
d,l‐Bornyl acetate	76‐49‐3	09.017	1.69	0.05–3.15
Myrcene	123‐35‐3	01.008	1.44	0.32–3.07
γ‐Terpinene	99‐85‐4	01.020	1.00	0.69–1.49
Linalool	78‐70‐6	02.013	0.97	0.63–1.51
Verbenone (Pin‐2‐en‐4‐one)	80‐57‐9	07.196	0.93	0.27–2.10
d,l‐Isobornyl acetate	125‐12‐2	09.218	0.74	0.70–0.80
4‐Terpinenol	562‐74‐3	02.072	0.66	0.02–1.22
α‐Fenchene	471‐84‐1	‐	0.58	0.03–1.68
(E)‐3,7‐Dimethylocta‐1,5,7‐trien‐3‐ol	53834‐70‐1	02.146	0.54	0.54–0.54
3,7,10‐Humulatriene	6753‐98‐6	01.043	0.49	0.09.1.15
Longifolene	475‐20‐7	01.047	0.46	0.44–0.48
α‐Terpinene	99‐86‐5	01.019	0.39	0.08–0.80
δ‐3‐Carene	13466‐78‐9	01.029	0.34	0.06–0.66
Terpinolene	586‐62‐9	01.005	0.34	0.06–0.73
α‐Phellandrene	99‐83‐2	01.006	0.31	0.19–0.52
α‐Thujene	2867‐05‐2	–	0.27	0.02–0.74
γ‐Terpineol	586‐81‐2	–	0.22	0.22–0.23
(+)‐δ‐Cadinene	483‐76‐1	–	0.20	0.03–0.40
Octan‐3‐one	106‐68‐3	07.062	0.19	0.06–0.56
Oct‐1‐en‐3‐ol	3391‐86‐4	02.023	0.17	0.13–0.21
trans‐Sabinene hydrate	17699‐16‐0	–	0.17	0.12–0.24
α‐Copaene	3856‐25‐5	–	0.15	0.03–0.36
γ‐Muurolene	30021‐74‐0	–	0.15	0.01–0.25
Tricyclene (1,1,7‐trimethyltricyclo[2.2.1.0.(2.6)]heptane)	508‐32‐7	01.060	0.13	0.01–0.26
β‐Caryophyllene epoxide	1139‐30‐6	16.043	0.12	0.02–0.30
γ‐Cadinene	39029‐41‐9	–	0.12	0.00–0.17
Pseudolimonene	499‐97‐8	–	0.11	0.11 3
Pinocamphone	547‐60‐4	–	0.11	0.01–0.45
γ‐Amorphene	6980‐46‐7	–	0.10	0.08–0.13
Sabinene (4(10)‐thujene)	3387‐41‐5	01.059	0.10	0.02–0.27
Total			98.4 4	94.4–99.8 5

TABLE 3. Maximum proposed use levels of rosemary oil in complete feed.

Animal category	Maximum proposed use level (mg/kg complete feed)
Chickens for fattening	300
Laying hens	300
Turkeys for fattening	100
Piglets	100
Pigs for fattening	100
Sows	100
Veal calves (milk replacers)	100
Cattle for fattening	500
Dairy cows	200
Sheep/goats	100
Horses	300
Rabbits	100
Fish (salmon)	300
Crustaceans (shrimps)	300
Dogs	130
Cats	100
Ornamental fish	100
Other species	300

TABLE 4. Flavouring components of rosemary oil already assessed by EFSA and/or by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) as chemically defined flavourings, grouped according to the chemical group (CG) as defined in Commission Regulation (EC) No 1565/2000, with indication of the EU Flavour Information System (FLAVIS) number and the corresponding EFSA/JECFA opinion.

CG	Chemical group	Product (EU register name)	FLAVIS No	EFSA/JECFA opinion,* Year
01	Straight‐chain primary aliphatic alcohols/aldehydes/acids, acetals and esters with esters containing saturated alcohols and acetals containing saturated aldehydes	Nonanal	05.025	2013
04	Non‐conjugated and accumulated unsaturated straight‐chain and branched‐chain aliphatic primary alcohols, aldehydes, acids, acetals and esters	Hex‐3(cis)‐en‐1‐ol	02.056	2016a
05	Saturated and unsaturated aliphatic secondary alcohols, ketones and esters with esters containing secondary alcohols	Oct‐1‐en‐3‐ol	02.023	2020a
		Isopulegol	02.067	2020a
		Octan‐3‐one	07.062	2015a, 2023b
06	Aliphatic, alicyclic and aromatic saturated and unsaturated tertiary alcohols and esters with esters containing tertiary alcohols ethers	Linalool	02.013	2012a, 2020b
		α‐Terpineol	02.014	2012a
		2‐(4‐Methylphenyl)propan‐2‐ol	02.042
		4‐Terpinenol	02.072
		(E)‐3,7‐Dimethylocta‐1,5,7‐trien‐3‐ol 1	02.146	WHO, 2002, (JECFA)
07	Primary alicyclic saturated and unsaturated alcohols/aldehydes/acids/acetals/esters with esters containing alicyclic alcohols	Myrtenol 1	02.091	2017, CEF
		2‐(2,2,3‐Trimethylcyclopent‐3‐enyl)ethan‐1‐ol 1 (α‐campholenol)	02.114	WHO, 2002, (JECFA)
08	Secondary alicyclic saturated and unsaturated alcohols, ketones, ketals and esters with ketals containing alicyclic alcohols or ketones and esters containing secondary alicyclic alcohols	d,l‐Fenchyl alcohol	02.038	2016b
		d,l‐Borneol	02.016	2016b, 2023c
		d,l‐Isoborneol	02.059
		d,l‐Bornyl acetate	09.017
		d,l‐Isobornyl acetate	09.218
		d‐Camphor 2	07.215
		p‐Menth‐1‐en‐3‐ol	02.083	2011a, CEF WHO, 2000a, (JECFA)
		Pinocarveol 1 , 3	02.100	2014a, CEF
		Pin‐2‐en‐4‐ol 1 , 3 (verbenol)	02.101
		Pin‐2‐en‐4‐one 1 , 4 (verbenone)	07.196	2012, CEF
09	Primary aliphatic saturated or unsaturated alcohols/aldehydes/acids/acetals/esters with a second primary, secondary or tertiary oxygenated functional group including aliphatic lactones	Ethyl brassylate 1	09.533	WHO, 2000b (JECFA)
10	Secondary aliphatic saturated or unsaturated alcohols, ketones, ketals and esters with a second secondary or tertiary oxygenated functional group	4‐Hydroxy‐4‐methylpentan‐2‐one 1	07.165	2011b, CEF
16	Aliphatic and alicyclic ethers	1,8‐Cineole	03.001	2012b, 2021b
18	Allylhydroxybenzenes	Eugenol	04.003	2011
25	Phenol derivatives containing ring‐alkyl, ring‐alkoxy and side‐chains with an oxygenated functional group	Thymol	04.006	2012c
		Carvacrol	04.031
30	Miscellaneous substances	Methyl 3‐oxo‐2‐pent‐2‐enyl‐1‐cyclopentylacetate 1 , 5	09.521	WHO, 2005, (JECFA)
31	Aliphatic and aromatic hydrocarbons and acetals containing saturated aldehydes	1‐Isopropyl‐4‐methylbenzene (p‐cymene)	01.002	2015b
		Terpinolene	01.005
		α‐Phellandrene	01.006
		α‐Terpinene	01.019
		d‐Limonene	01.045
		γ‐Terpinene	01.020
		Pin‐2(10)‐ene (β‐pinene)	01.003	2016c
		Pin‐2(3)‐ene (α‐pinene)	01.004
		β‐Caryophyllene	01.007
		Myrcene	01.008
		Camphene	01.009
		δ‐3‐Carene	01.029
		Germacra‐1(10),4(14),5‐triene δ‐Germacrene 1 , 6	01.042	2011c, CEF
		3,7,10‐Humulatriene 1 , 6	01.043
		Longifolene 1 , 6	01.047
		α‐Muurolene 1 , 6	01.052
		1,1,7‐trimethyltricyclo [2.2.1.0.(2.6)]heptane (tricyclene) 1 , 6	01.060
		β‐Bisabolene 1	01.028	2015a, CEF
		4(10)‐Thujene (sabinene) 1	01.059
		cis‐3,7‐Dimethyl‐1,3,6‐octatriene (cis‐β‐ocimene) 1 , 7	01.064
32	Epoxides	β‐Caryophyllene epoxide 1	16.043	2014b, CEF

TABLE 5. Concentration of camphor in complete feed resulting from the use of rosemary oil at the maximum proposed use levels and calculated maximum safe concentrations of rosemary oil in complete feed (mg/kg) to ensure a safe level of camphor (5 mg/kg) for the different target animal categories.

Animal category	Daily feed intake (g DM/kg bw)	Proposed use level of rosemary oil (mg/kg complete feed) 1	Concentration of camphor (mg/kg complete feed) 2	Maximum safe use level of rosemary oil (mg/kg complete feed) 1 , 3
Chickens for fattening	79	300	72	20.8
Laying hens	53	300	72	20.8
Turkeys for fattening	59	100	24	20.8
Pig for fattening	44	100	24	20.8
Piglets	37	100	24	20.8
Sows lactating	30	100	24	20.8
Veal calves (milk replacer)	19	100	24	20.8
Cattle for fattening	20	500	120	20.8
Dairy cows	31	200	48	20.8
Sheep/goats	20	100	24	20.8
Horses	20	300	72	20.8
Rabbits	50	100	24	20.8
Salmonids	18	300	72	20.8
Dogs	17	300	72	20.8
Cats	20	130	31.2	20.8
Ornamental fish	5	100	24	20.8

TABLE 6. Compositional data, intake values (calculated for chickens for fattening at 20.8 mg/kg complete feed), reference points, margin of exposure (MOE) for the individual components of rosemary oil classified according to assessment groups, and combined margin of exposure (MOET) for each assessment group.

Essential oil composition			Exposure		Hazard characterisation		Risk characterisation
Assessment group	FLAVIS‐No	Highest concentration in the oil	Highest concentration in feed	Daily intake 1	Cramer class 2	NOAEL 3	MOE 4	MOET 5
Constituent	–	%	mg/kg	mg/kg bw/day	–	mg/kg bw/day	–	–
CG 5
Oct‐1‐en‐3‐ol	02.023	0.21	0.044	0.0039	(I)	6.7	1709
Isopulegol	02.067	0.06	0.012	0.0010	(I)	38	36,340
MOET CG 5								1632
CG 6
α‐Terpineol	02.014	2.70	0.562	0.0504	(I)	125 6	2479
4‐Terpinenol	02.072	1.22	0.254	0.0228	(I)	125 6	5474
(E)‐3,7‐Dimethylocta‐1,5,7‐trien‐3‐ol	–	0.54	0.113	0.0101	(I)	44	4356
trans‐Sabinene hydrate	–	0.24	0.050	0.0045	(I)	222	49,745
γ‐Terpineol	–	0.23	0.047	0.0042	(I)	125 6	29,752
2‐(4‐Methylphenyl)propan‐2‐ol	–	0.05	0.011	0.0010	I	3	3031
MOET CG 6								688
CG 7
Myrtenol	02.091	0.09	0.018	0.0016	I	3	1868
2‐(2,2,3‐Trimethylcyclopent‐3‐enyl)ethan‐1‐ol	02.114	0.05	0.011	0.0010	I	3	3150
MOET CG 7								1173
CG 8
Verbenone	07.196	2.10	0.437	0.0392	(II)	60	1530
Pinocamphone	–	0.45	0.094	0.0084	(II)	60	7109
Pinocarvone	–	0.14	0.028	0.0025	(II)	60	23,802
Fenchyl alcohol	02.038	0.08	0.017	0.0016	I	3	1913
MOET CG 8								736
CG 9
Ethyl brassylate	09.533	0.02	0.005	0.0004	I	3	7303
CG 10
4‐Hydroxy‐4‐methylpentan‐2‐one	13.095	0.03	0.007	0.0006	I	3	4725
CG 16
1,8‐Cineole	03.001	62	12.896	1.1577	(II)	100	86
CG 30
Methyl 3‐oxo‐2‐pent‐2‐enyl‐1‐cyclopentylacetate	09.521	0.02	0.004	0.0004	II	0.15	2437
CG 31, II (Acyclic alkanes)
Myrcene	01.008	1.37	0.639	0.0573	(I)	44	768
cis‐β‐Ocimene	01.064	0.06	0.018	0.0016	(I)	44	27,400
trans‐β‐Ocimene	–	0.05	0.018	0.0016	(I)	44	27,085
trans‐β‐Farnesene	–	0.06	0.017	0.0015	(I)	44	29,455
Group MOE CG 31, II							709
CG 31, III (Cyclohexene hydrocarbons)
d‐Limonene	01.046	4.13	0.860	0.0772	(I)	250	3239
γ‐Terpinene	01.020	1.49	0.310	0.0278	(I)	250	8998
α‐Terpinene	01.019	0.80	0.166	0.0149	(I)	30 7	2011
Terpinolene	01.005	0.73	0.152	0.0136	(I)	250	18,315
α‐Phellandrene	01.006	0.52	0.108	0.0097	(I)	30 7	3090
Pseudolimonene	–	0.11	0.023	0.0021	I	3	1461
m‐Camphorene	–	0.04	0.008	0.0007	I	3	4017
MOET CG 31,III								449
CG 31, IVe (Benzene hydrocarbons, alkyl)
p‐Cymene	01.002	2.95	0.614	0.0551	(I)	154	2796
Abietatriene	–	0.04	0.008	0.0007	I	3	4017
β‐Calacorene	–	0.04	0.008	0.0007	I	3	4017
α‐Calacorene	–	0.01	0.002	0.0001	I	3	20,083
MOET CG 31, IVe								1104
CG 31, V (Bi‐, tricyclic, non‐aromatic hydrocarbons)
α‐Pinene	01.004	26.00	5.408	0.4855	(I)	222	457
Camphene	01.009	15.00	3.120	0.2801	(I)	222	793
β‐Pinene	01.003	12.00	2.496	0.2241	(I)	222	991
β‐Caryophyllene	01.007	5.61	1.166	0.1047	(I)	222	2121
α‐Fenchene	–	1.68	0.349	0.0313	(I)	222	7089
α‐Thujene	–	0.74	0.154	0.0138	(I)	222	16,066
δ‐3‐Carene	01.029	0.66	0.137	0.0123	(I)	222	18,014
Longifolene	01.047	0.48	0.100	0.0090	(I)	222	24,769
(+)‐δ‐Cadinene	01.021	0.40	0.084	0.0075	(I)	222	29,501
α‐Copaene	–	0.36	0.076	0.0068	(I)	222	32,752
Sabinene	01.059	0.27	0.055	0.0050	(I)	222	44,695
Tricyclene	01.060	0.26	0.055	0.0049	(I)	222	45,034
γ‐Muurolene	–	0.25	0.051	0.0046	(I)	222	48,134
2,4‐Thujadiene	–	0.10	0.022	0.0019	I	3	1545
Isocadinene	–	0.03	0.006	0.0005	I	3	5738
Cadina‐1,4‐diene	–	0.03	0.006	0.0005	I	3	5950
MOET CG 31, V								158
CG 31, VI (Macrocyclic non‐aromatic hydrocarbons)
3,7,10‐Humulatriene	01.043	1.15	0.239	0.0215	(I)	111 8	5169
Germacra‐1(10),4(14),5‐triene	01.042	0.05	0.010	0.0009	I	3	3213
MOET CG 31, VI								1982
CG 32
β‐Caryophyllene epoxide	16.043	0.30	0.062	0.0056	(III)	109	19,458

TABLE 7. Margin of exposure (MOE) for 1,8‐cineole (CG 16) calculated for the different target animal categories at the safe concentration of 20.8 mg rosemary oil/kg complete feed (calculated considering the content of camphor in rosemary oil) and maximum safe use level in complete feed.

Animal category	Daily feed intake (g DM/kg bw)	Lowest MOE CG 16	Maximum safe use level (mg/kg complete feed) 1
Chickens for fattening	79	86	18
Laying hens	53	128	20.8
Turkeys for fattening	59	115	20.8
Pigs for fattening	44	154	20.8
Piglets	37	184	20.8
Sows lactating	30	226	20.8
Veal calves (milk replacer)	19	358	20.8
Cattle for fattening	20	340	20.8
Dairy cows	31	219	20.8
Sheep/goats	20	340	20.8
Horses	20	340	20.8
Rabbits	50	136	20.8
Salmonids	18	377	20.8
Dogs	17	400	20.8
Cats 2	20	340	14.1
Ornamental fish	5	1359	20.8

TABLE 8. Individual intake of methyleugenol and estragole, their combined intake and group margin of exposure (MOE) calculated for the target animal categories at the reduced safe concentrations reported in Table 7.

Target species	Daily feed intake	Use level in feed 1	Methyleugenol intake 2	Estragole intake 2	Combined intake 2	MOE 3 , 4
	g DM/kg bw	mg/kg	μg/kg bw per day
Long‐living and reproductive animals
Laying hens	53	20.8	0.489	0.338	0.827	26,851
Piglets	44	20.8	0.406	0.281	0.686	32,343
Sows lactating	30	20.8	0.278	0.193	0.471	47,166
Dairy cows	31	20.8	0.284	0.196	0.480	46,250
Sheep/goats	20	20.8	0.184	0.128	0.312	71,154
Horses	20	20.8	0.184	0.128	0.312	71,154
Rabbits	50	20.8	0.461	0.319	0.780	28,462
Dogs	17	20.8	0.154	0.106	0.260	85,385
Cats	20	14.1	0.125	0.087	0.212	104,965
Ornamental fish	5	20.8	0.041	0.029	0.070	316,239
Target species for fattening
Chickens for fattening	79	18	0.630	0.436	1.067	9376
Turkeys for fattening	59	20.8	0.541	0.374	0.915	10,927
Pigs for fattening	37	20.8	0.338	0.234	0.572	17,483
Veal calves (milk replacer)	19	20.8	0.162	0.112	0.275	36,422
Cattle for fattening	20	20.8	0.184	0.128	0.312	32,051
Sheep/goats	20	20.8	0.184	0.128	0.312	32,051
Horses for fattening	20	20.8	0.184	0.128	0.312	32,051
Rabbits	50	20.8	0.461	0.319	0.780	12,821
Salmonids	18	20.8	0.161	0.112	0.273	36,630

TABLE 9. Concentrations of rosemary oil in complete feed (mg/kg) considered very unlikely to induce adverse effects in long‐living and reproductive animals and of no concern for target species for fattening.

Animal categories	Concentration (mg/kg complete feed) 1
Turkeys for fattening	20.8
Chickens for fattening and minor poultry for fattening	18.0
Laying hens and other laying/reproductive birds including animals reared for laying/reproduction and ornamental birds	20.8
Piglets and piglets of minor porcine species including animals reared for reproduction	20.8
Pigs for fattening and other porcine species for meat production	20.8
Sows and other porcine species for reproduction	20.8
Veal calves (milk replacer)	20.8
Sheep/goats	20.8
Cattle for fattening, other ruminants for fattening and camelids at the same physiological stage	20.8
Dairy cows and other ruminants and camelids for milk production or reproduction including animals reared for milk production/reproduction	20.8
Horses and other equids	20.8
Rabbits and other leporids	20.8
Salmonids and minor fin fish	20.8
Dogs	20.8
Cats	14.1
Ornamental fish	20.8
Other species	14.1

Full text

INTRODUCTION

1

Background and Terms of Reference

1.1

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. 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 Feed Flavourings Authorisation Consortium European Economic Interest Grouping (FFAC EEIG)2 for authorisation/re‐evaluation of 41 additives (king of bitter extract, thyme leaved gratiola tincture, devil's claw extract, devil's claw tincture, lavender oil, lavender tincture, spike lavender oil, melissa oil, balm leaves extract, mentha arvensis/corn mint oil, pennyroyal oil, spearmint oil, peppermint oil, peppermint tincture, basil oil, basil tincture, olive extract, marjoram oil, oregano oil, oregano tincture, patchouli oil, rosemary oil, rosemary oleoresin, rosemary extract, rosemary tincture, Spanish sage oil, sage oil, sage tincture, clary sage oil, savoury summer oil, savoury summer tincture, Pau darco tincture, thymus origanum oil, thyme oil, thyme oleoresin, thyme extract, thyme tincture, lilac chastetree extract, lilac chastetree tincture, Spanish marjoram oil and wild thyme tincture) belonging to botanically defined group (BDG) 01 – Lamiales, when used as a feed additive for all animal species (category: sensory additives; functional group: flavouring compounds). During the assessment, the applicant withdrew the applications for 12 additives.3 These additives were deleted from the register of feed additives.4 In addition, during the course of the assessment, the application was split and the present opinion covers only one out of the remaining 29 additives under application: rosemary oil for use in all animal species.

The remaining 28 additives belonging to the botanically defined group (BDG) 01 – Lamiales, under application are assessed in separate opinions.

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 1 June 2011.

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 feed additive consisting of rosemary oil when used under the proposed conditions of use (see Section 3.3.3).

Additional information

1.2

Rosemary oil from Rosmarinus officinalis L. is currently authorised as a feed additive according to the entry in the European Union Register of Feed Additives pursuant to Regulation (EC) No 1831/2003 (2b natural products – botanically defined). It has not been assessed as a feed additive in the EU. It should be noted that the designation Rosmarinus officinalis L. is no longer accepted by the botanical community and the species has been moved to the genus Salvia with the species name Salvia rosmarinus Spenn. However, Rosmarinus officinalis L. is used in the opinion when citing literature (e.g. PhEur, EMA monographs) and the EFSA Compendium on botanicals.

DATA AND METHODOLOGIES

2

Data

2.1

The present assessment is based on data submitted by the applicant in the form of a technical dossier5 in support of the authorisation request for the use of rosemary oil from S. rosmarinus as feed additives. The dossier was received on 26 June 2025 and the general information and supporting documentation are available at https://open.efsa.europa.eu/questions/EFSA‐Q‐2025‐00402.6

The FEEDAP Panel 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 already been evaluated by the FEEDAP Panel as chemically defined flavourings (CDGs). The applicant submitted a written agreement to reuse the data submitted for the assessment of chemically defined flavourings (dossiers, publications and unpublished reports) for the risk assessment of preparations belonging to BDG 01, including the current ones under assessment.7

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 additives. During the assessment, upon request of EFSA, the EURL issued two partial reports.8 The additive under assessment is included in the second partial report. In particular, for the characterisation of rosemary oil, the EURL recommended methods based on a method based on gas chromatography with flame ionisation detection (GC‐FID) for the quantification of the phytochemical markers 1,8‐cineole and camphor in rosemary oil.9

Methodologies

2.2

The approach followed by the FEEDAP Panel to assess the safety and the efficacy of rosemary oil from S. rosmarinus 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 Scientific Committee, 2009); Compendium of botanicals;11 Guidance on the identity, characterisation and conditions of use of feed additives (EFSA FEEDAP Panel, 2017a); Guidance on the safety of feed additives for the target species (EFSA FEEDAP Panel, 2017b); Guidance on the assessment of the safety of feed additives for the consumer (EFSA FEEDAP Panel, 2017c); Guidance on the assessment of the safety of feed additives for the environment (EFSA FEEDAP Panel, 2019); Guidance on the assessment of the safety of feed additives for the users (EFSA FEEDAP Panel, 2023a); Guidance on the assessment of the efficacy of feed additives (EFSA FEEDAP Panel, 2024); Guidance document on harmonised methodologies for human health, animal health and ecological risk assessment of combined exposure to multiple chemicals (EFSA Scientific Committee, 2019a); Statement on the genotoxicity assessment of chemical mixtures (EFSA Scientific Committee, 2019b); Guidance on the use of the Threshold of Toxicological Concern approach in food safety assessment (EFSA Scientific Committee, 2019c); General approach to assess the safety for the target species of botanical preparations which contain compounds that are genotoxic and/or carcinogenic (EFSA FEEDAP Panel, 2021a).12

ASSESSMENT

3

The additive under assessment, rosemary oil, is an essential oil obtained from the fresh or dried flowering aerial parts of Salvia rosmarinus Spenn. and is intended for use as a sensory additive (functional group: flavouring compounds) in feed and water for drinking for all animal species.

Origin and extraction

3.1

Salvia rosmarinus Spenn. (homotypic syn. Rosmarinus officinalis L.), commonly referred to as rosemary, is an evergreen shrub belonging to the Lamiaceae family. It is native to the Mediterranean region and introduced into more northern parts of Europe and to southern parts of North America because of its ornamental value and the use of its leaves as a culinary herb. Rosemary may be found in many forms either growing with an upright habit (1–2 m in height) or as a trailing or creeping plant. It is characterised by its needle‐like leaves, green on the upper and white on the lower surface. Flowers in season are randomly distributed along the stem and may be white, purple or blue in colour.

Following an extensive phylogenetic study of the genus Salvia and related genera (Drew et al., 2017), the taxonomic standing of the genus Rosmarinus has been revised and the species described for this genus moved to the genus Salvia. As a consequence, Rosmarinus is no longer considered an accepted genus although the synonym for S. rosmarinus (Rosmarinus officinalis) may still be found associated with descriptions involving the rosemary plant and its extracts.

The additive is extracted from the fresh or dried flowering aerial parts of S. rosmarinus by steam distillation. The volatile constituents are condensed and then separated from the aqueous phase by decantation.

Uses other than feed flavourings

3.2

While there is no specific EU authorisation for any preparation described as coming from S. rosmarinus when used to provide flavour in food, according to Regulation (EC) No 1334/2008,13 flavouring preparations produced from food 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.’

‘Rosemary leaf (Rosmarini folium)’ and ‘Rosemary oil (Rosmarini aetheroleum)’ from R. officinalis L. are described in monographs of the European Pharmacopoeia 11.0 (PhEur, 2022a, 2022b) and of the European Medicines Agency (EMA, 2024a, 2024b, 2024c) for medicinal uses.

Characterisation

3.3

Characterisation of rosemary oil

3.3.1

The essential oil is obtained from the flowering aerial parts of Salvia rosmarinus Spenn. sourced from Spain, Tunisia, Morocco and India. It is a clear, pale‐yellow liquid with a strong woody and herbaceous note. Rosemary oil is identified with the single Chemical Abstracts Service (CAS) number 8000‐25‐7, the European Community (EC) number 283‐291‐9,14 the Flavor Extract Manufacturers Association (FEMA) number 2992 and the Council of Europe (CoE) number 406. In eight batches of the additive, the refractive index (20°C) fell within the range of 1.466–1.469. In six batches, the density (20°C) ranged between 901 and 911 kg/m^3^, and the optical rotation (20°C) between −0.88° and −0.25° in five batches.15

The product specifications used by the applicant are based on the standard developed by the International Organisation for Standardization (ISO) 1342:2012 (‘Essential oil of rosemary [Rosmarinus officinalis L.]’) and the European Pharmacopoeia (Eur. Ph.) 01/2008:1846 (‘Rosemary oil, (Rosmarini aetheroleum) [Rosmarinus officinalis L.]’)16 adapted to reflect the concentrations of selected volatile components. Five components contribute to the specifications as shown in Table 1, with 1,8‐cineole and camphor selected as the phytochemical markers. Analysis of 10 batches of the additive showed compliance with the specifications when analysed by GC‐FID and expressed as a percentage of gas chromatographic peak area (% GC area).17

The applicant provided a full analysis of 13 batches using gas chromatography–mass spectrometry (GC–MS).18 In total, up to 87 peaks were detected and identified in the chromatograms, accounting for on average 99.1% (94.8%–100%) of the GC area. The five specified compounds accounted for 76.5% on average (67.8%–90.0%) of the GC area when measured by GC–MS (Table 1). Besides the five compounds indicated in the product specifications, 36 compounds were detected at individual levels of > 0.1% and are listed in Table 2. These 41 compounds together account on average for 98.4% (range 94.8%–99.8%) of the GC area. The remaining 46 compounds (ranging between 0.1% and 0.008%) and accounting for on average 0.65% of the GC area are listed in the footnote.19 Based on the available data, rosemary oil is considered a fully characterised mixture, as defined by the EFSA Scientific Committee (2019a).

An analysis of rosemary oil under assessment detected low concentrations of methyleugenol (0.010%–0.039%) in six batches and estragole (0.020%–0.027%) in three batches.

The EFSA Compendium of botanicals20 reports as substances of potential concern for human and animal health the occurrence of 1,8‐cineole (22.01%–52.2%), camphor (7.57%–10.08%), l‐piperitone (6.68%), myrcene (1.63%–2.61%) and verbenone (23.55%) in the essential oil from the aerial parts of R. officinalis.

The applicant carried out an extensive literature search on the chemical composition of R. officinalis and its botanical preparations and the possible presence of substances of concern.21 Several cumulative databases (including LIVIVO, NCBI, OVID and ToxInfo) were searched. The keywords used covered different aspects of safety, and the inclusion and exclusion criteria were provided by the applicant. The literature search (no time limits specified) retrieved 67 references investigating the composition of preparations from R. officinalis. The results of the literature search confirmed the information in the EFSA Compendium. In addition, a few publications reported the presence of 0.9%–2.0% methyleugenol (Hazarika et al., 2020; Song et al., 2023) and 0.13% estragole (Dolghi et al., 2022; Moussii et al., 2020) in essential oils from R. officinalis. No other substances of concern were identified in the literature provided by the applicant.

Impurities

3.3.1.1

The applicant referred to the ‘periodic testing’ of some representative flavourings premixtures for mercury, cadmium, lead, arsenic, fluoride, dioxins and polychlorinated biphenyls (PCBs), organo‐chlorine pesticides, organo‐phosphorous pesticides, aflatoxins (B1, B2, G1, G2) and ochratoxin A. However, no data were provided on the presence of these impurities.

Shelf‐life

3.3.2

The applicant states that the typical shelf‐life of rosemary oil is at least 12 months when stored in tightly closed containers under standard conditions (in a cool, dry place protected from light).22 However, no data supporting this statement were provided.

Conditions of use

3.3.3

Rosemary oil is intended to be added to feed and water for drinking for all animal species without a withdrawal period. The maximum proposed use levels in complete feed are listed in Table 3. No use level has been proposed for the use of the additive in water for drinking.

Safety

3.4

No studies to support the safety for target animals, consumers and users were performed with the additive under assessment.

The applicant provided a literature search on the genotoxicity of preparations obtained from the leaves of S. rosmarinus. An in vivo study performed in rodents administered by gavage with 300, 1000, 2000 mg/kg body weight (bw) of rosemary oil (composition not given) showed genotoxic effects in bone marrow erythrocytes (Maistro et al., 2010). However, since no information was reported on the composition of the test item, the FEEDAP Panel did not consider this in vivo study relevant for the assessment.

The publication by Maistro et al. (2010) is also referenced in the EFSA Compendium on botanicals, together with another study (Contini et al., 2020) reporting positive results in an in vitro micronucleus assay. Since the positive response was associated with a high level of cytotoxicity, the FEEDAP Panel considers these effects with caution, in line with OECD TG 487 recommendations. The genotoxicity of rosemary oil was reviewed in the FEMA GRAS assessment of natural flavour (Eisenbrand et al., 2021) and in the Cosmetic Ingredient Review Expert Panel (Panel) assessment (Fiume et al., 2018). The FEEDAP Panel notes that negative results of genotoxicity testing reported in both assessments for rosemary oil were based on unpublished reports, which were not made available by the applicant. Therefore, none of the data could be considered as relevant for the assessment of the genotoxicity of rosemary oil.

In line with the recommendation of the EFSA Scientific Committee for the genotoxicity assessment of fully defined chemical mixtures (EFSA Scientific Committee, 2019b), the assessment of genotoxicity is based on the identified components.

Forty‐nine of the individual components of the essential oil, accounting on average for 82.3% of the GC area, have already been assessed as chemically defined flavourings for use in feed and food by the FEEDAP Panel, the EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids (CEF) and/or by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). The flavouring compounds together with the EU Flavour Information System (FLAVIS) number, the chemical group as defined in Commission Regulation (EC) No 1565/2000,23 and the corresponding EFSA opinion are listed in Table 4. Camphor (as a mixture of isomers) has not been evaluated for use as a flavouring but is closely related to the flavouring compound *d‐*camphor [07.215] already assessed by EFSA for use in food and feed. d‐Camphor is therefore included in Table 4.

Camphor is present in the additive as a mixture of enantiomers (d,l‐camphor, the ratio between d‐ and *l‐*stereoisomers is not given) and accounts on average for 15.3% of the GC area.

Five compounds listed in Table 4, δ‐germacrene [01.042], 3,7,10‐humulatriene [01.043], longifolene [01.047], α‐muurolene [01.052] and tricyclene [01.060], have been evaluated in Flavouring Group Evaluations 25 Revision 2 by applying the procedure described in the Guidance on the data required for the risk assessment of flavourings to be used in or on foods (EFSA CEF Panel, 2010). For these compounds, for which there is no concern for genotoxicity, EFSA requested additional subchronic toxicity data (EFSA CEF Panel, 2011b). In the absence of these data, the CEF Panel was unable to complete its assessment (EFSA CEF Panel, 2015a). As a result, these compounds are no longer authorised for use as flavours in food. For these compounds, in the absence of toxicity data, the FEEDAP Panel applies the threshold of toxicological concern (TTC) approach or read‐across from structurally related substances, as recommended in the Guidance document on harmonised methodologies for human health, animal health and ecological risk assessment of combined exposure to multiple chemicals (EFSA Scientific Committee, 2019a).

The 44 remaining components of rosemary oil listed in Table 4, accounting on average for about 81.6% of the GC area, are considered safe for use as flavourings. They are currently authorised for use in food24 without limitations and for use in feed25 at individual use levels higher than those resulting from the intended use in feed of the essential oil under assessment.

*d‐*Camphor [07.147], *d,l‐*borneol [02.016] and d,l‐isobornyl acetate [09.218] were included in tolerance studies made with the mixture of flavourings referred to as ‘Herbal’ (EFSA FEEDAP Panel, 2023c). Linalool [02.013] and octan‐3‐one [07.062] were similarly tested in mixtures of flavourings named ‘TuttiFrutti’ and ‘MilkyVanilla’, respectively (EFSA FEEDAP Panel, 2020b, 2023b). Based on these studies, the FEEDAP Panel concluded that d,l‐borneol [02.016] is safe at 15 mg/kg complete feed for all animal species, *d,l‐*isobornyl acetate [09.218] and d‐camphor [07.147] at 5 mg/kg, linalool [02.013] at 30 mg/kg, and octan‐3‐one [07.062] at 10 mg/kg. The FEEDAP Panel considered that the conclusions reached for d‐camphor [07.147] can be extrapolated to the mixture of isomers of camphor. By applying read‐across, the FEEDAP Panel also concluded that *d,l‐*bornyl acetate [09.017] and *d,l‐*isoborneol [02.059] are safe for the target species at 5 mg/kg complete feed (EFSA FEEDAP Panel, 2023c).

Thirty‐five volatile compounds have not been previously assessed for use as flavourings. The FEEDAP Panel notes that 34 of them,26 accounting on average for 1.4% of the GC area, are aliphatic monoterpenes or sesquiterpenes structurally related to flavourings already assessed in CG 6, CG 8 and CG 31, and a similar metabolic and toxicological profile is expected. Because of their lipophilic nature, they are expected to be rapidly absorbed from the gastro‐intestinal tract, oxidised to polar oxygenated metabolites, conjugated and excreted, and not to accumulate in animal tissues and products (EFSA FEEDAP Panel, 2012a, 2015b, 2016b, 2016c). Humulene oxide II is structurally related to β‐caryophyllene oxide in CG 32, and read‐across is applied (EFSA CEF Panel, 2014b).

Six batches of the additive contained low concentrations of methyleugenol (0.010%–0.039%) and three batches contained estragole (0.020%–0.027%).

For the absorption, distribution, metabolism and excretion (ADME) and the toxicology of methyleugenol and estragole, reference is made to the safety evaluation made by the FEEDAP Panel in the opinion on laurel leaf oil (EFSA FEEDAP Panel, 2023).

Safety for the target species

3.4.1

Tolerance studies in the target species and/or toxicological studies in laboratory animals with the essential oil under assessment were not submitted. In the absence of these data, the approach to the safety assessment of a mixture whose individual components are known is based on the safety assessment of each individual component (component‐based approach). This approach requires that the mixture is sufficiently characterised and that the individual components can be grouped into assessment groups, based on structural and metabolic similarity. The combined toxicity can be predicted using the dose addition assumption within an assessment group, taking into account the relative toxic potency of each component (EFSA Scientific Committee, 2019a).

As the additive under assessment is a fully defined mixture (the identified components represent on average 99.1% of the GC area, see Section 3.3.1), the FEEDAP Panel applied a component‐based approach to assess the safety for target species of the essential oil.

The oil under assessment contains by specification up to 24% of camphor, which is assessed separately from the other components of the oil, based on the results of tolerance trials with the mixture of flavourings ‘Herbal’ (EFSA FEEDAP Panel, 2023c). Similarly, linalool [02.013], octan‐3‐one [07.062], d,l‐borneol [02.016] and d,l‐isobornyl acetate [09.218], which were included in tolerance studies with the mixtures of flavourings ‘TuttiFrutti’, ‘MilkyVanilla’ and ‘Herbal’ (EFSA FEEDAP Panel, 2020b, 2023b, 2023c), are assessed separately. The structurally related compounds d,l‐isoborneol [02.059] and d,l‐bornyl acetate [09.017] are assessed together with d,l‐borneol [02.016] and d,l‐isobornyl acetate [09.218]. The remaining identified components are assessed using the component‐based approach.

Methyleugenol and estragole, substances for which a concern for genotoxicity has been identified, are assessed separately. For these compounds, belonging to the group of p‐allylalkoxybenzenes, rodent carcinogenicity studies with methyleugenol are available from which a benchmark dose (BMD) lower confidence limit for a benchmark response of 10% (BMDL_10_) can be derived (EFSA FEEDAP Panel, 2023).

Components tested in tolerance trials with mixtures of flavourings Camphor

3.4.1.1

At the proposed use levels for rosemary oil, the concentration of camphor would range from 24 to 120 mg/kg complete feed, considering that camphor is present in the essential oil under assessment at the highest specification of 24% (see Table 5).

The levels to which animals would be exposed using the additive at the highest proposed use levels would exceed the level of 5 mg camphor/kg complete feed considered safe in a previous opinion (EFSA FEEDAP Panel, 2023c). The Panel recalculated the safe level of the additive as 20.8 mg rosemary oil/kg complete feed considering the presence of camphor in rosemary oil.

Other components were tested in tolerance trials with mixtures of flavourings.

The highest concentrations in feed of octan‐3‐one [07.062] resulting from the use of the additive at the maximum proposed use levels would be 0.56–2.8 mg/kg complete feed.27 These concentrations are below the safe level of 10 mg/kg complete feed established for all animal species based on the results of the tolerance trials with the mixture of flavourings ‘MilkyVanilla’ (EFSA FEEDAP Panel, 2023b).

The highest concentrations in feed of linalool [02.013] resulting from the use of the additive at the maximum proposed use levels would be 1.51–7.55 mg/kg complete feed.28 These concentrations are below the safe level of 30 mg/kg complete feed established by the tolerance trials with the mixture of flavourings ‘TuttiFrutti’ (EFSA FEEDAP Panel, 2020b).

When considering the highest analysed concentrations in rosemary oil, the four (iso)bornyl derivatives (d,l‐borneol [02.016], d,l‐isoborneol [02.059], d,l‐bornyl acetate [09.017], d,l‐isobornyl acetate [09.218]) would account together for up to 10.3% of the GC area. The highest feed concentration for the sum of these derivatives resulting from the use of the rosemary oil at the proposed use levels (100–500 mg/kg complete feed) would range between 10.3 and 51.5 mg/kg complete feed,29 which is above the safe level of 15 mg/kg complete feed experimentally established for *d,l‐*borneol in tolerance trials with the mixture of flavourings ‘Herbal’ and of 5 mg/kg for the other (iso)bornyl derivatives established in tolerance trials or by applying read‐across from d,l‐isobornyl acetate. However, at the concentration of 20.8 mg rosemary oil/kg complete feed calculated considering the content of camphor (see Table 4), the resulting feed concentration of (iso)bornyl derivatives would be 2.14 mg/kg complete feed, which is considered safe for all animal species.

Components other than those tested in tolerance trials

3.4.1.2

Based on considerations related to structural and metabolic similarities, the remaining components were allocated to 12 assessment groups, corresponding to the chemical groups (CGs) 1, 4, 5, 6, 8, 16, 18, 23, 25, 30, 31 and 32, as defined in Annex I of Regulation (EC) No 1565/2000. For CG 31 (aliphatic and aromatic hydrocarbons), sub‐assessment groups as defined in Flavouring Group Evaluation 25 (FGE.25) and FGE.78 were established (EFSA CEF Panel, 2015a, 2015b). The allocation of the components to the (sub‐)assessment groups is shown in Table 6 and in the corresponding footnote.

For hazard characterisation, each component of an assessment group was first assigned to the structural class according to Cramer classification using Toxtree (version 3.1.0, May 201830). For some components in the assessment group, toxicological data were available to identify no observed adverse effect levels (NOAELs). Structural and metabolic similarity among the components in the assessment groups was assessed to explore the application of read‐across, allowing extrapolation from a known NOAEL of a component of an assessment group to the other components of the group with no available NOAEL or, if sufficient evidence were available for members of a (sub‐)assessment group, to derive a (sub‐)assessment group NOAEL.

Subchronic studies from which NOAEL values could be identified were available for the following components: 127 mg/kg bw per day for hex‐3*(cis)‐*en‐1‐ol [02.056] in CG 4 (EFSA FEEDAP Panel, 2016a), 38 mg/kg bw per day for isopulegol [02.067] and 6.7 mg/kg bw per day for oct‐1‐en‐3‐one [07.081] in CG 5 (EFSA FEEDAP Panel, 2020a), 250 mg/kg bw per day for terpineol [02.230]31 in CG 6 (EFSA FEEDAP Panel, 2012a), 100 mg/kg bw per day for 1,8‐cineole [03.001] in CG 16 (EFSA FEEDAP Panel, 2021b), 300 mg/kg bw per day for eugenol [04.004] in CG 18 (EFSA FEEDAP Panel, 2011), 44 mg/kg bw per day for myrcene [01.008], 154 mg/kg bw per day for p‐cymene [01.002], 250 mg/kg bw per day for d‐limonene [01.046] and 222 mg/kg bw per day for β‐caryophyllene [01.007] in CG 31 (EFSA FEEDAP Panel, 2015b, 2016c) and 109 mg/kg bw per day for β‐caryophyllene epoxide [16.043] in CG 32 (EFSA CEF Panel, 2014b). For α‐terpinene [01.019], the FEEDAP Panel identified a NOAEL of 60 mg/kg bw per day based on maternal toxicity (reduced body weight gain) in a teratogenicity study in rats (Araujo et al., 1996; also reported in ECHA, 2018). The NOAEL of 60 mg/kg bw per day was divided by a factor of 2 to take account of the nature of the study.

For CG 1, a group NOAEL of 120 mg/kg per day was identified from the toxicological data available for several compounds (EFSA FEEDAP Panel, 2013), which is applied to nonanal [05.025] in the current assessment.

In CG 5, the NOAEL of 6.7 mg/kg bw per day for oct‐1‐en‐3‐one [07.081] was applied to oct‐1‐en‐3‐ol [02.023].

In CG 6, a NOAEL of 250 mg/kg bw per day was identified for terpineol [02.230] and divided by a factor of 2 to take account of the nature of the study (EFSA FEEDAP Panel, 2012a). The resulting value of 125 mg/kg bw per day was used as the reference point for terpinyl derivatives, α‐terpineol [02.014], 4‐terpinenol [02.072], γ‐terpineol and trans‐p‐2‐menthen‐1‐ol.

In CG 8, the BMDL_10_ of 60 mg/kg bw per day for d‐carvone (EFSA Scientific Committee, 2014) was applied to the structurally related ketones pinocarvone, verbenone [07.196] and pinocamphone.32

Since a compound‐specific NOAEL has been identified for α‐terpinene [01.019], which is lower than that of d‐limonene [01.045], the representative compound in CG 31, III, the FEEDAP Panel considered the need to review the read‐across applied within this group. The assessment group ‘cyclohexene derivatives’ includes compounds characterised by the presence of at least two double bonds, which can be either isolated (as in d‐limonene) or conjugated (as in α‐terpinene). For the two subgroups of compounds, a refinement in read‐across is applied as follows: the NOAEL of 250 mg/kg bw per day for d‐limonene is applied to the compounds with isolated double bonds and the NOAEL of 60 mg/kg bw per day for α‐terpinene to the compounds with conjugated double bonds.

Considering the structural and metabolic similarities, the NOAELs for the representative compounds in CG 31, myrcene [01.008], d‐limonene [01.045] and β‐caryophyllene [01.007] were applied, respectively, using read‐across to the compounds within sub‐assessment groups II (cis‐β‐ocimene [01.064], trans‐β‐ocimene, trans‐β‐farnesene, α‐farnesene), III (γ‐terpinene [01.020], terpinolene [01.005] and β‐bisabolene [01.028]), and V (α‐pinene [01.004], camphene [01.009], β‐pinene [01.003], α‐fenchene, α‐thujene, δ‐3‐carene [01.029], longifolene [01.047], (+)‐δ‐cadinene, α‐copaene, sabinene [01.059], tricyclene [01.060], γ‐muurolene, γ‐cadinene, γ‐amorphene, α‐ylanglene, β‐copaene, α‐cubebene, α‐muurolene [01.052], aromadendrene, α‐amorphene, α‐longipinene, β‐selinene, isocaryophyllene, α‐cadinene, δ‐selinene),33 respectively (EFSA CEF Panel, 2015a, 2015b). In the current assessment, the NOAEL of 60 mg/kg bw per day identified for α‐terpinene [01.019] and divided by a factor of 2 to take account of the nature of the study is applied to α‐phellandrene [01.006].

The NOAEL of 44 mg/kg bw per day for myrcene [01.008] was also applied to *(E)‐*3,7‐dimethylocta‐1,5,7‐trien‐3‐ol in CG 6.34 Similarly, the NOAEL of 222 mg/kg bw per day for β‐caryophyllene [01.007] was also applied to trans‐sabinene hydrate and camphene hydrate in CG 6, pinocarveol [02.100] and verbenol [02.101] in CG 8, and 3,7,10‐humulatriene [01.043] in CG 31, VI.35 For 3,7,10‐humulatriene, the NOAEL of β‐caryophyllene was divided by a factor of 2 to take into account of the uncertainty in read‐across due to differences in the structure (extrapolation from a tricyclic to a macrocyclic non‐aromatic compound) (EFSA FEEDAP Panel, 2023).

For the remaining compounds,36 toxicity studies performed with the compounds under assessment were not available and read‐across was not possible. Therefore, the threshold of toxicological concern (TTC) approach was applied (EFSA FEEDAP Panel, 2017b; EFSA Scientific Committee, 2019c).

As the result of the hazard characterisation, a reference point was identified for each component in the assessment group based on the toxicity data available (NOAEL from in vivo toxicity study or read‐across) or from the 5th percentile of the distribution of NOAELs of the corresponding Cramer Class (i.e. 3 mg/kg bw per day for Cramer Class I compounds, Munro et al., 1996). Reference points selected for each compound are shown in Table 6.

For each component in the assessment group, exposure in target animals (expressed as mg/kg bw per day) was estimated considering the maximum proposed use levels of the additive in feed, the percentage of the component in the oil and the default values for body weight and feed intake according to the guidance on the safety of feed additives for target species (EFSA FEEDAP Panel, 2017b). For the compounds covered by specification (1,8‐cineole, α‐pinene, camphene and β‐pinene, see Table 1), the indicated maximum specified concentration is used for the calculation of exposure. For the other components, the highest analysed concentration is used.

For risk characterisation, the margin of exposure (MOE) was calculated for each component as the ratio between the reference point and the exposure. For an assessment group, (i) when a group reference point is available, a group MOE was calculated for the combined intake; (ii) when different reference points are available for the components, the combined (total) margin of exposure (MOET) was calculated as the reciprocal of the sum of the reciprocals of the MOE for the individual substances (EFSA Scientific Committee, 2019a). An MOE(T) > 100 allowed for interspecies differences and intra‐individual variability. The volatile compounds resulting individually in an MOE > 50,000 were not further considered in the assessment group as their contribution to the MOE(T) is negligible. They are listed in the footnote.37

The approach to the safety assessment of rosemary oil is shown in Table 6 for chickens for fattening, which is the species with the highest ratio of feed intake/body weight and represents the worst‐case scenario among the target species. The calculations were done at the maximum use level of 20.8 mg/kg complete feed, considered safe based on the presence of camphor.

As shown in Table 6, for all assessment groups, except CG 16, the MOE(T) calculated for chickens for fattening at the safe concentration calculated considering the content of camphor (20.8 mg/kg complete feed) was > 100. The lowest MOE of 86 was calculated for 1,8‐cineole (CG 16). The MOEs for CG 16 compounds calculated for the other target species at 20.8 mg/kg complete feed are summarised in Table 7.

At the use level of 20.8 mg/kg complete feed, the MOE exceeds the value of 100 for all animal species except for chickens for fattening. For chickens for fattening, the maximum safe use level in feed was calculated to ensure an MOE ≥ 100. Because glucuronidation is an important metabolic pathway to facilitate the excretion of the components of the essential oil and considering that cats have an unusually low capacity for glucuronidation, particularly for aromatic compounds (Court & Greenblatt, 1997; Lautz et al., 2021), the use of rosemary oil as an additive in cat feed needs a wider margin of exposure. An MOE of 500 is considered adequate.

For all the animal species and categories listed in Table 7 except chickens for fattening and cats, rosemary oil is considered safe (without considering the presence of methyleugenol and estragole) when used as a feed additive at 20.8 mg/kg complete feed. For chickens for fattening and cats, the resulting maximum safe levels in feed (without considering the presence of methyleugenol and estragole) are shown in Table 7.

Methyleugenol and estragole

3.4.1.3

Methyleugenol (0.010%–0.039%) was detected in six batches and estragole (0.020%–0.027%) was detected in three batches.

Methyleugenol and estragole belong to the group of p‐allylalkoxybenzenes and are genotoxic and carcinogenic. According to the General approach to assess the safety for the target species of botanical preparations which contain compounds that are genotoxic and/or carcinogenic (EFSA FEEDAP Panel, 2021a), different reference points and a different magnitude of the margin of exposure (MOE) are applied for long‐living and reproductive animals (including those animals reared for laying/reproduction) and for short‐living animals. Short‐living animals are defined as those animals raised for fattening whose lifespan under farming conditions makes it very unlikely that they develop cancer as a result of the exposure to genotoxic and/or carcinogenic substances in the diet.

For long‐living and reproductive animals, an MOE with a magnitude > 10,000 when comparing estimated exposure to genotoxic and/or carcinogenic substances with a BMDL_10_ from a rodent carcinogenicity study is considered indicative of low concern. The FEEDAP Panel identified the BMDL_10_ of 22.2 mg/kg bw per day derived from carcinogenicity studies in rodents (rats) with methyleugenol (NTP, 2000; Suparmi et al., 2019), as the reference point for the entire group of *p‐*allylalkoxybenzenes (EFSA FEEDAP Panel, 2022).

For short‐living animals, genotoxicity and carcinogenicity endpoints are not considered biologically relevant; therefore, a lower magnitude of the MOE (> 100) when comparing estimated exposure with a reference point based on non‐neoplastic endpoints is considered adequate (EFSA FEEDAP Panel, 2021b). The FEEDAP Panel identified a NOAEL of 10 mg/kg bw per day for non‐neoplastic lesions (changes in organ weight38 and function, including effects on the liver39 and the glandular stomach40) from a 90‐day study in mice with methyleugenol (EFSA FEEDAP Panel, 2023; NTP, 2000).

The highest daily combined intake of methyleugenol and estragole was calculated considering the maximum safe use levels of the additive in feed for the different animal categories (Table 7) and the maximum analysed concentrations of methyleugenol and estragole in the additive. The intake values are reported in Table 8, together with the corresponding group MOE for the combined intake. This is calculated considering the relevant reference point for long‐living and reproductive animals and for short‐living animals (species for fattening).

When the estimated exposures of long‐living animals are compared to the BMDL_10_ of 22.2 mg/kg bw per day derived for methyleugenol by Suparmi et al. (2019) from a rodent carcinogenicity study (NTP, 2000), an MOE > 10,000 is obtained for long‐living and reproductive animals (Table 8). In accordance with the EFSA Statement on the margin of exposure (EFSA Scientific Committee, 2012), the use of an additive in feed is considered of low concern for long‐living and reproductive animals if the MOE is > 10,000. Consequently, the FEEDAP Panel considers it very unlikely that the use of the feed additive will induce adverse effects during their lifetime.

For short‐living animals (species for fattening), the magnitude of the MOE is > 100 (Table 8) and is of no safety concern when comparing the exposure to the reference point of 10 mg methyleugenol/kg bw per day for non‐neoplastic endpoints.

Overall evaluation

3.4.1.4

When considering the assessment of the individual components or groups of components of rosemary oil, the maximum concentrations of the additive in feed were calculated based on the presence of camphor and, for chickens for fattening and cats, also considering the presence of 1,8‐cineole (see Table 7). These concentrations are considered of no safety concern for short‐living animals (species for fattening) and, with regard to the presence of methyleugenol and estragole, are considered to be very unlikely to induce adverse effects in long‐living and reproductive animals. These levels are extrapolated to physiologically related species. For the other species not considered, the calculated maximum level of 14.1 mg additive/kg complete feed is applied.

No specific use levels have been proposed by the applicant for the use level of rosemary oil in water for drinking. The FEEDAP Panel considers that the use of the additive in water for drinking alone or in combination with use in feed should not exceed the daily amount that is considered very unlikely to induce adverse effects in long‐living and reproductive animals and of no safety concern for short‐living animals (species for fattening) when consumed via feed.

Conclusions on the safety for the target species

3.4.1.5

The FEEDAP Panel considers that the use of rosemary oil is very unlikely to induce adverse effects in long‐living and reproductive animals and is of no safety concern for short‐living animals (species for fattening) up to the maximum feed concentrations summarised in Table 9.

The FEEDAP Panel considers that the use of the additive in water for drinking alone or in combination with use in feed should not exceed the daily amount that is considered very unlikely to induce adverse effects in long‐living and reproductive animals and of no safety concern for short‐living animals (species for fattening) when consumed via feed.

Safety for the consumer

3.4.2

Rosemary and its preparations, including rosemary oil, are added to a wide range of food categories as spice or for flavouring purposes. Although individual consumption figures for the EU are not available, Fenaroli's handbook of flavour ingredients (Burdock, 2009) cites values of 0.44 mg/kg bw per day for rosemary and its extracts and 0.003 mg/kg bw per day for rosemary oil obtained from the fresh flowering tops.

Most of the individual constituents of the essential oil under assessment are currently authorised as food flavourings without limitations and have already been assessed for consumer safety when used as feed additives in animal production (see Table 4, section 3.4).

No data on residues in products of animal origin were made available for any of the constituents of the essential oil. However, the Panel recognises that the constituents of rosemary oil are expected to be extensively metabolised and excreted in the target species. For the major components, 1,8‐cineole, camphor, α‐pinene, camphene and β‐pinene, the available data in laboratory animals and humans indicate that they are absorbed, metabolised by oxidation and conjugation, and excreted; they are not expected to accumulate in animal tissues and products (EFSA FEEDAP Panel, 2012b, 2016b, 2016c). Consequently, relevant residues in food products are unlikely.

The physiologically based kinetic (PBK) model simulations in target species indicated that the carry‐over of methyleugenol and estragole and their corresponding 1'‐hydroxy metabolites to tissues and products (e.g. eggs, milk) is limited due to rapid elimination and a decline in concentrations in edible tissues (Noorlander et al., 2024). Since methyleugenol and estragole occur in the additive at trace concentrations, relevant residues in food products of animal origin are unlikely.

Considering the above and the reported human exposure due to the direct use of rosemary and its preparations in food (Burdock, 2009), the FEEDAP Panel considers that consumption of products from animals given rosemary oil at the maximum use level proposed by the applicant would be unlikely to significantly increase human background exposure.

No safety concern would be expected for consumers of animal products from the use of rosemary oil under the proposed conditions of use.

Safety for the user

3.4.3

No specific data were provided by the applicant regarding the safety of the additive for users.

The applicant made a literature search aimed at retrieving studies related to the safety of preparations obtained from S. rosmarinus for users.41 None of the references retrieved were considered relevant to the safety assessment.

The applicant provided a safety data sheet42 for rosemary oil, which identified concerns for dermal and eye, and dermal and respiratory sensitisation. The FEEDAP Panel concludes that rosemary oil should be considered as irritant to skin and eyes, and as a dermal and respiratory sensitiser. When handling rosemary oil, exposure of unprotected users to methyleugenol and estragole may occur. Therefore, to reduce the risk, the exposure of the users should be minimised.

Safety for the environment

3.4.4

S. rosmarinus is a species native to Europe where it is widely grown both for commercial and decorative purposes. Therefore, the use of the rosemary oil under the proposed conditions of use in animal feed is not expected to pose a risk to the environment.

Efficacy

3.5

Rosemary (described under the synonym R. officinalis) and its oil are listed in Fenaroli's Handbook of Flavour Ingredients (Burdock, 2009) and by the Flavour and Extract Manufacturers Association (FEMA) with the reference numbers 2991 and 2992, respectively.

Since rosemary 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 for rosemary oil.

CONCLUSIONS

4

Rosemary oil from Salvia rosmarinus Spenn (homotypic syn. Rosmarinus officinalis L.) may be produced from plants of different origins and by various processes resulting in preparations with different composition and toxicological profiles. Thus, the following conclusions apply only to rosemary oil which contains ≤ 0.039% methyleugenol and ≤ 0.027% estragole and is produced from the flowering aerial parts of S. rosmarinus.

The concentrations in complete feed of rosemary oil, which are considered very unlikely to induce adverse effects in long‐living and reproductive animals and of no safety concern for short‐living animals (species for fattening) are summarised as following:Animal categoriesConcentration (mg/kg complete feed)1 Turkeys for fattening20.8Chickens for fattening and minor poultry for fattening18.0Laying hens and other laying/reproductive birds including animals reared for laying/reproduction and ornamental birds20.8Piglets and piglets of minor porcine species including animals reared for reproduction20.8Pigs for fattening and other porcine species for meat production20.8Sows and other porcine species for reproduction20.8Veal calves (milk replacer)20.8Sheep/goats20.8Cattle for fattening, other ruminants for fattening and camelids at the same physiological stage20.8Dairy cows and other ruminants and camelids for milk production or reproduction including animals reared for milk production/reproduction20.8Horses and other equids20.8Rabbits20.8Salmonids and minor fin fish20.8Dogs20.8Cats14.1Ornamental fish20.8Other species14.1 ^1^ Complete feed containing 88% dry matter, milk replacer 94.5% dry matter.

The FEEDAP Panel considers that the use in water for drinking alone or in combination with use in feed should not exceed the daily amount that is considered very unlikely to induce adverse effects in long‐living and reproductive animals and of no safety concern for short‐living animals (species for fattening) when consumed via feed.

No safety concerns were identified for the consumer and the environment from the use of the additive in animal feed.

The additive under assessment should be considered as irritant to skin and eyes, and as dermal and respiratory sensitiser. When handling rosemary oil, exposure of unprotected users to methyleugenol and estragole may occur. Therefore, to reduce the risk, the exposure of the users should be minimised.

Since S. rosmarinus 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.

RECOMMENDATION

5

The specifications should ensure that rosemary oil contains ≤ 0.039% methyleugenol and ≤ 0.027% estragole.

DOCUMENTATION PROVIDED TO EFSA/CHRONOLOGY

6

DateEvent 23/11/2010 Dossier received by EFSA. Botanically defined flavourings from Botanical Group 01 – Lamiales for all animal species and categories. Submitted by Feed Flavourings Authorisation Consortium European Economic Interest Grouping (FFAC EEIG) 03/01/2011 Reception mandate from the European Commission 06/01/2011 Application validated by EFSA – Start of the scientific assessment 01/04/2011 Request of supplementary information to the applicant in line with Article 8(1)(2) of Regulation (EC) No 1831/2003 – Scientific assessment suspended. Issues: analytical methods

08/01/2013 Reception of supplementary information from the applicant ‐ Scientific assessment remains suspended 26/02/2013 EFSA informed the applicant (EFSA ref. 7150727) that, in view of the workload, the evaluation of applications on feed flavourings would be re‐organised by giving priority to the assessment of the chemically defined feed flavourings, as agreed with the European Commission 24/06/2015 Technical hearing during risk assessment with the applicant according to the “EFSA's Catalogue of support initiatives during the life‐cycle of applications for regulated products”: data requirement for the risk assessment of botanicals 27/02/2019 Partial withdrawal by applicant (EC was informed) for the following additives: Thyme leaves gratiola tincture, spike lavender oil, melissa oil, pennyroyal oil, basil oil and savory summer oil 30/06/2021 EFSA informed the applicant that the evaluation process restarted 08/07/2021 Request of supplementary information to the applicant in line with Article 8(1)(2) of Regulation (EC) No 1831/2003 – Scientific assessment suspended. Issues: characterisation, safety for target species, safety for the consumer, safety for the user and environment

28/09/2023 Partial withdrawal of the application for the following additive: Spanish majoram oil 08/07/2024 Partial withdrawal of the application for the following additives: lilac chastetree extract and savory summer tincture 26/08/2024 Reception of the Evaluation report of the European Union Reference Laboratory for Feed Additives. Partial report related to seven additives: Spanish sage oil, peppermint oil, thymus origanum oil, patchouli oil, clary sage oil, lavender oil and sage oil 27/08/2024 Reception of supplementary information from the applicant (letter of agreement) 16/12/2024 Partial withdrawal of the application for the following additives: devils claw extract (wb), balm leaves extract (sb), olive extract (sb) 08/01/2025 Reception of supplementary information from the applicant (partial dataset: rosemary oil) ‐ Scientific assessment remains suspended 18/06/2025 Reception of the Evaluation report of the European Union Reference Laboratory for Feed Additives. Partial report related to 11 additives: cornmint oil, spearmint oil, thyme oil, rosemary oil, marjoram oil, rosemary tincture, basil tincture, lavender tincture, peppermint tincture, sage tincture and wild thyme tincture 26/06/2025 The application was split and a new EFSA‐Q‐2025‐00402 was assigned to the additive included in the present assessment. Scientific assessment re‐started for the additive included in the present assessment 16/09/2025 Opinion adopted by the FEEDAP Panel on rosemary oil (EFSA‐Q‐2025‐00402). End of the Scientific assessment for the additive included in the present assessment. The assessment of other additives in BGD 01 is still ongoing

ABBREVIATIONSADMEAbsorption, distribution, metabolism and excretionAFCEFSA Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with FoodBDGbotanically defined groupBMDBenchmark doseBMDL_10_ benchmark dose (BMD) lower confidence limit for a benchmark response of 10%BWbody weightCASChemical Abstracts ServiceCEFEFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing AidsCDGchemically defined groupCGchemical groupCLPClassification, Labelling and PackagingCoECouncil of EuropeDMdry matterECHAEuropean Chemicals AgencyEEIGEuropean economic interest groupingEINECSEuropean Inventory of Existing Chemical SubstancesEMAEuropean Medicines AgencyEURLEuropean Union Reference LaboratoryFEMAFlavour and Extract Manufactures AssociationFEEDAPEFSA Scientific Panel on Additives and Products or Substances used in Animal FeedFFACFeed Flavourings authorisation Consortium of FEFANA (EU Association of Specialty Feed Ingredients and their Mixtures)FGEflavouring group evaluationFLAVISThe EU Flavour Information SystemFL‐noFLAVIS numberJECFAJoint FAO/WHO Expert Committee of Food AdditivesGC‐FIDgas chromatography‐flame ionisation detectionGC–MSgas chromatography–mass spectrometryISOInternational Organisation for StandardisationLODlimit of detectionMOEmargin of exposureMOETcombined margin of exposure (total)NOAELno observed adverse effect levelNTPNational Toxicology ProgramOECDOrganisation for Economic Co‐operation and DevelopmentQSARQuantitative Structure Activity RelationshipPhEurEuropean PharmacopoeiaSCEFSA Scientific CommitteeTTCthreshold of toxicological concernUFuncertainty factorWHOWorld Health Organization

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2010‐01037 (New EFSA‐Q‐2025‐00402)

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PANEL MEMBERS

Roberto Edoardo Villa, Giovanna Azimonti, Eleftherios Bonos, Henrik Christensen, Mojca Durjava, Birgit Dusemund, Ronette Gehring, Boet Glandorf, Maryline Kouba, Marta López‐Alonso, Francesca Marcon, Carlo Nebbia, Alena Pechová, Miguel Prieto‐Maradona, Ilen Röhe, and Katerina Theodoridou.