Safety of Lycopene oleoresin from tomatoes

The EFSA Journal (2008) 675, 1-22

Safety of Lycopene oleoresin from tomatoes 1
Scientific Opinion of the Panel on Scientific Panel on Dietetic Products,
Nutrition and Allergies
(Question No EFSA-Q-2006-186)
Adopted on 24 April 2008

PANEL MEMBERS
Jean-Louis Bresson, Albert Flynn, Marina Heinonen, Karin Hulshof, Hannu Korhonen, Pagona
Lagiou, Martinus Løvik, Rosangela Marchelli, Ambroise Martin, Bevan Moseley, Andreu
Palou, Hildegard Przyrembel, Seppo Salminen, John Sean Strain, Stephan Strobel, Inge Tetens,
Henk van den Berg, Hendrik van Loveren, and Hans Verhagen.

SUMMARY
Following a request from European Commission, the Panel on Dietetic Products, Nutrition and
Allergies was asked to deliver a scientific opinion on the safety of ‘lycopene oleoresin from
tomatoes’ for use as a novel food ingredient.

The novel food ingredient consists of a lycopene-rich oleoresin obtained as an ethyl acetate
extract from the pulp of ripe tomatoes from a non-GM variety of tomatoes (Lycopersicon
lycopersicum L. Karst. ex Farw) naturally selected for their high lycopene content.

Lycopene extracted from tomatoes is authorised within the EU as food colouring agent
(E160d). The lycopene oleoresin from tomatoes to be used as a novel food ingredient contains
5-15 % lycopene and is prepared by a production process identical to that for the production of
the additive E160d, although E160d is prepared using an additional concentration step to obtain
an oleoresin that contains 60-70 % lycopene.

The identity and purity of the lycopene oleoresin from tomatoes is in compliance with the
purity criteria for colouring agent E160d (lycopene) as food additive.
The applicant proposes to use the lycopene oleoresin as a food ingredient. Products will be
formulated in such a way that they will provide about 2 mg lycopene per serving. Lycopene

1 For citation purposes: Scientific Opinion of the Panel on Panel on Dietetic Products, Nutrition and Allergies on a request
from the European Commission on the safety of ‘lycopene oleoresin from tomatoes’. The EFSA Journal (2008) 675, 1-22.
© European Food Safety Authority, 2008

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Safety of Lycopene Oleoresin from Tomatoes

oleoresin as a food supplement has been used in the UK since before 1997 to a significant degree
and thus does not need further approval.
The Panel based its intake estimates for lycopene on four sources including 1) normal dietary
intake of lycopene from food, 2) intake of lycopene from dietary supplements, 3) intake of
lycopene from proposed food products and 4) use of lycopene as a food colour.
An overview of average dietary intakes of lycopene from foods in different populations was
presented in previous EFSA evaluations. It was concluded that regular intakes of lycopene
from natural dietary sources in different populations are, according to dietary surveys,
estimated to be on average between 0.5 and 5 mg/day, with high exposures up to about 8
mg/day. High consumption of fruits and vegetables, especially tomato products, may result in
occasional intakes of 20 mg lycopene/day or more.
The applicant indicates that the amount of lycopene used in dietary supplements is 5 to 15
mg/day.

The applicant estimates that the use of lycopene at the proposed use levels in the six food
categories requested would lead to an overall additional lycopene intake of 12 mg per day.
In a recent opinion the Scientific Panel on Food Additives, Flavourings, Processing Aids and
Materials in Contact with Food noted that total daily exposure to lycopene as a food colour
could potentially range from 2 to 6 mg on average and up to 23 mg at the high level.
The AFC Panel did not exclude an occasionally combined high exposure from both natural
dietary sources and food colours of up to 43 mg of lycopene per day.
Overall, the NDA Panel concludes that intake of lycopene from the proposed levels of use as a
novel food ingredient would lead to intake levels that would substantially increase the overall
dietary intake of lycopene, and could lead to mean daily intakes from 14.5 - 23 mg/day and from
46 - 70 mg/day as high intakes. These values amount to 0.24 - 0.38 mg/kg bw/day and to 0.77 -
1.17 mg/kg bw/day for a 60 kg person and are for the high intake estimates substantially higher
than the acceptable daily intake (ADI) recently established by the AFC Panel as a group ADI of
0.5 mg/kg bw/day for lycopene from all sources (EFSA, 2008). This ADI is in line with the
ADI of 0 - 0.5 mg/kg bw/day established by JECFA (JECFA, 2006).
The Panel considers that lycopene oleoresin from tomatoes is as safe as lycopene from other
accepted sources.
The Panel concludes that for the average user consumption of lycopene from tomatoes oleoresin
and from all other sources will be below the ADI. However, considering the various sources of
lycopene (natural occurrence, fortified foods, supplements, food colour) some users of lycopene
products may exceed the ADI of 0.5 mg/kg bw/day.


Key words:
Lycopene oleoresin from tomatoes, novel food ingredient, E160d, Registry Number 502-65-8

The EFSA Journal (2008) 675, 2-22

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Safety of Lycopene Oleoresin from Tomatoes

TABLE OF CONTENTS
Panel Members............................................................................................................................................1
Summary .....................................................................................................................................................1
Table of Contents ........................................................................................................................................3
Background as provided by the commission...............................................................................................4
Terms of reference as provided by the commission....................................................................................6
Acknowledgements .....................................................................................................................................6
Assessment ..................................................................................................................................................7
I.
Specification of the novel food (NF)..............................................................................................7
II. Effect of the production process applied to the NF ........................................................................9
III.
History of the organism used as the source of the NF....................................................................9
IX.
Anticipated intake/extent of use of the NF.....................................................................................9
X.
Information from previous human exposure to the NF or its source ...........................................11
XI.
Nutritional information on the NF ...............................................................................................12
XII.
Microbiological information on the NF .......................................................................................14
XIII. Toxicological information on the NF...........................................................................................14
Discussion .................................................................................................................................................17
Conclusions and Recommendations..........................................................................................................18
Documentation provided to EFSA ............................................................................................................18
References .................................................................................................................................................18

The EFSA Journal (2008) 675, 3-22

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Safety of Lycopene Oleoresin from Tomatoes

BACKGROUND AS PROVIDED BY THE COMMISSION
On 7 September 2004, Barry Ottaway & Associates Ltd. submitted a request under Article 4 of
the Novel Food Regulation (EC) No 258/97 to the Competent Authorities of the United
Kingdom for placing on the market ‘lycopene oleoresin from tomatoes’ Lyc-O-Mato® as a
novel food ingredient.

On 30 June 2005, the Competent Authorities of the United Kingdom forwarded to the
Commission their initial assessment report, which had reached the conclusion that ‘lycopene
oleoresin from tomatoes’ for the proposed uses is acceptable, subject to adherence to the
proposed specification and the production parameters.

On 9 August 2005, the Commission forwarded the initial assessment report to the other
Member States. Several of these Member States submitted additional comments/objections. In
consequence, a Community Decision is now required under Article 7, paragraph 1 of
Regulation (EC) No 258/97.

The concerns of scientific nature raised by the Competent Authorities of Member States can be
summarised as follows:

• Details on the production process, the absence of harmful substances and the
specifications of the final product and the tomato source should be clarified.
• Lycopene is also authorised as a food additive. The proposed levels for use as a novel
food ingredient are higher and for more food categories than those for use as a food
colour. It should be guaranteed that the authorisation as a novel food ingredient would
not lead to its use as a colouring agent without permission.
• Toxicological data are insufficient to guarantee the safety of intake levels above the
normal dietary intake. Due to the large range of products for which use levels are
foreseen the exposure will increase. A complete risk assessment of the intake of
lycopene from all sources should be made.
• In a 13-week oral toxicity study in rats, there was a significant increase in lung weight
in females. Arguments to support that this effect is not of toxicological concern should
be provided and/or, given the absence of adequate mutagenicity data, a long-term study
should be required.
• There should be clarification regarding the nutritional benefit of adding lycopene to the
diet
• It would be important to present studies regarding certain factors that could potentially
affect the bio-availability of lycopene, as well as an evaluation of its interaction with
other carotenoids present in the diet, in order to assess the possible differences between
this product and foods naturally rich in lycopene.
• It is important to protect allergic individuals and prevent small children from over-
consumption of tomato lycopene.

In addressing Member States’ comments of a scientific nature, and considering the overall
safety of lycopene oleoresin from tomatoes the Panel has used information from the original
dossier provided by the applicant, the initial assessment carried out by the authorities of the

The EFSA Journal (2008) 675, 4-22

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Safety of Lycopene Oleoresin from Tomatoes

United Kingdom, the comments given by the Member States, the response from the applicant
to the issues raised by the Member States and the recent opinion of the Scientific Panel on
Food Additives, Flavourings, Processing Aids and Materials in Contact with Food on the use of
lycopene as a food additive (EFSA, 2008).

Existing authorisations and evaluations
Lycopene, extracted from tomatoes, is authorised as a food colouring agent within the EU (E160d)
(Directive 94/36/EC) and the US (CDR 21 73.295).
Lycopene was evaluated by the SCF in 1975 when it was unable to allocate an ADI but felt
able to accept the use of lycopene prepared from natural foods by physical processes, without
further investigations, as a colouring matter in food, provided that the amount consumed did
not differ significantly from the amount consumed through the relevant foodstuffs (SCF, 1975).
This Opinion was reiterated in 1989 (SCF, 1989). When JECFA evaluated lycopene from
natural sources in 1977 they postponed a decision because of lack of data (JECFA, 1978).
In 1999 the SCF evaluated synthetic lycopene, but the available data were not sufficient to be
accepted by the SCF. The SCF concluded (SCF, 1999): “The proposed specification ‘not less
than 96 %’ lycopene is not acceptable because highly concentrated lycopene is sensitive to
oxygen and light, forms degradation products with mutagenic activity, and is not identical with
the beadlet formulation that has been tested toxicologically,” and “The toxicological data
provided on the beadlet formulation are insufficient. Therefore the Committee is not able to
allocate an ADI and considers its use in food unacceptable at present.”
Synthetic lycopene is currently not approved for colouring matters within the EU. It is
considered generally recognised as safe (GRAS) for use as a food ingredient in the US (GRAS
notice No GRN 000119).

Recently the Panel evaluated the use of an ?-tocopherol-containing oil suspension of lycopene
obtained from Blakeslea trispora for use as a novel food (EFSA, 2005). It was concluded that an
?-tocopherol-containing oil suspension of lycopene obtained from B. trispora to an additional
intake of up to about 2 mg/day is not of concern from the safety point of view. It was also
concluded that this does not hold for the proposed levels of use of lycopene in foods that would
give rise to an additional intake of 20 mg per day.

In July 2006, the Joint FAO/WHO Expert Committee on Food Additives (JECFA, 2006)
derived an ADI of 0 - 0.5 mg/kg bw/day based on a 104-week study in rats using a safety factor
of 100 for synthetic lycopene. This ADI was made into a group ADI to include lycopene from
B. trispora. This level equates to 30 mg lycopene/day maximum for a 60 kg individual.

The Panel noted that the ADI set by JECFA (JECFA, 2006) does not include lycopene from
tomatoes. This was due to the fact that JECFA was not evaluating lycopene from tomatoes.

In a recent opinion the Scientific Panel on Food Additives, Flavourings, Processing Aids and
Materials in Contact with Food (EFSA, 2008) evaluated the new toxicological data on lycopene
and derived an ADI of 0.5 mg/kg bw/day. This ADI refers to lycopene from all sources.


The EFSA Journal (2008) 675, 5-22

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Safety of Lycopene Oleoresin from Tomatoes

TERMS OF REFERENCE AS PROVIDED BY THE COMMISSION
In accordance with Article 9 (1) (a) of Regulation (EC) No 178/2002, the European Food
Safety Authority is asked to carry out the additional assessment for ‘lycopene oleoresin from
tomatoes’ (Lyc-O-Mato®) in the context of Regulation (EC) No 258/97.

In particular, EFSA is asked to consider the elements of a scientific nature in the
comments/objections raised by the other Member States.

Furthermore, EFSA is asked to consider whether there are reasons to believe that ‘lycopene
oleoresin from tomatoes’ as specified by the applicant is less safe than lycopene stabilised by
other means.

ACKNOWLEDGEMENTS
The European Food Safety Authority wishes to thank the members of the Working Group for
the preparation of this opinion: Jean-Louis Bresson, Karl-Heinz Engel, Marina Heinonen,
Pagona Lagiou, Bevan Moseley, Andreu Palou, Annette Pöting, Seppo Salminen, Hendrik
Van Loveren, Hans Verhagen; and ad hoc expert Ivonne Rietjens.

The EFSA Journal (2008) 675, 6-22

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Safety of Lycopene Oleoresin from Tomatoes

ASSESSMENT
In accordance with the Commission Recommendation 97/618/EC, the ingredient concerned by
the application belongs to Class 2.1. “Complex NF from non-GM sources: the source of the NF
has a history of food use in the Community”. For this reason the opinion will be an assessment
of the safety data provided by the applicant to comply with the information required for novel
foods of Class 2.1, i.e. information requirements I, II, III, IX, X, XI, XII and XIII as detailed in
the following text. It does not include an assessment of the possible nutritional benefits of
lycopene oleoresin.

I. Specification of the novel food (NF)
The novel food ingredient (NI) (Lyc-O-Mato®) consists of a lycopene-rich oleoresin obtained
as an ethyl acetate extract from the pulp of ripe tomatoes from a non-GM hybrid variety
(Lycopersicon lycopersicum L. Karst. ex Farw) that has been selected for its naturally high
lycopene content.

Lycopene is a carotenoid with the formula C40H56. It has a molecular weight of 536.85 and the
CAS Registry Number 502-65-8. Its structural formula is:
H C
3
CH
CH
H C
CH
3
3
3
3
H C
CH
3
CH
CH
3
3
3
CH3
Fig 1. Structural Formula

Lycopene occurs in food predominantly in an all-trans form (Cronin, 2000; Boileau et al.,
2002). Regular tomatoes and tomato products mainly contain the all-E (trans-)isomers of
lycopene (between 35-96 % of total lycopene content), but also some Z (cis-)isomers, mainly
as 5Z, 9Z, 13Z and 15Z in percentages varying between 1-22 % (Schierle et al., 1997).

Carotenoids from tomato or tomato extract can be analysed using HPLC (Ishida et al., 2001).

Based on the analytical information provided by the applicant, the novel food ingredient
consists of 5-15 % lycopene together with a number of other constituents that occur naturally in
tomato, including fatty acids and acylglycerols (69-74 %), unsaponifiable matter (14-19 %),
water soluble matter (2.7-4.7 %), water (0.48-0.86 %), phosphorous compounds (0.35-0.52 %),
phospholipids (8.9-14 %), nitrogen (0.16-0.31 %) and sulphated ash (upon drying)(0.7-0.8 %).

Lycopene in the proposed novel food ingredient consists of 90-95 % (all-trans)-lycopene.
Different cis isomers are also likely to be present at small quantities. Other carotenoids and
related substances including ?-carotene, phytofluene, phytoene and tocopherols are, according
to the applicant, present in small amounts (0.1-2.5 %).

The applicant notes that the percentage of lycopene in the oleoresin may vary between 5 and
15 % because the composition of the tomatoes is subject to natural fluctuations. The analysis of
25 commercial batches produced between 1995 and 2003 revealed a range of 5.8 to 15.6 %.


The EFSA Journal (2008) 675, 7-22

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Safety of Lycopene Oleoresin from Tomatoes

Specifications for the lycopene oleoresin from tomatoes have been provided by the applicant
and are presented in Table 1. The applicant indicates that the levels of solvent residues,
pesticides, microbiological contamination and heavy metals are continuously monitored.

Table 1. Proposed Specifications of lycopene oleoresin from tomatoes as provided by the
applicant
Analysis Method
Specification
Observation against
Physical State
Red to dark brown viscous liquid
standard
Clarity LAB/123/01
Clear
solution
Lycopene identity
LAB/109/01a
HPLC retention time
Total lycopeneb
LAB/109/01a
5.0 to 15.0 %
% trans-Lycopene LAB/109/01a
90 to 95 %
Total carotenoidsc
LAB/102/02a
6.5 to 16.5 %
Phytoene: 0.5 to 0.75 %
Other carotenoids
LAB/118/01a
Phytofluene: 0.4 to 0.65 %
ß-carotene: 0.2 to 0.35 %
Total tocopherols
LAB/118/01
1.5 to 3.0 %
Unsaponifiable
Study 98/021a
13 to 20 %
matter
Total fatty acidsd
Study 98/021
60 to 75 %
Phytosterols
Study 98/021
0.5 to 2.5 %
90 % < 5?
Lycopene crystal particle size Microscopic
99 % < 10?
Water
Karl Fisher
0.5 % max
Sulphated ash
AOAC 34.104
0.5 to 1.5 %
Residual solvent (ethyl
LAB/114/01a
50 mg/kg max
acetate, ethanol)
Pesticides DFG-S19e

Below 3 ppm
Heavy metals
I.C.P.e
Pb < 2 mg/kg
Cd, Mo, Ni, Hg all <1 mg/kg
Arsenic I.C.P.e
As < 2 mg/kg
Total viable count
Moulds
Yeasts
< 1000/g
Escherichia coli:
< 100/g
Salmonella sp
< 100/g
Microbiology
USP 24 NF 19/<61>
Staphylococcus aureus Not detected in 10g
Pseudomonas
Not detected in 20g
aeruginosa
Not detected in 10g
Clostridium
Not detected in 10g
perfringens

aLycoRed Method, SOP available on request
bCombined cis- and trans-lycopenes
cCalculated as lycopene
dMyristic acid (14:0); palmitic acid (16:0); stearic acid (18:0); oleic acid (18:1); linoleic acid (18:2); linolenic acid
(18:3); arachidic acid (20:0); behenic acid (22:0); mono-, di- and tri-glycerides; free fatty acids
eAnalysis method SOP available on request


The EFSA Journal (2008) 675, 8-22

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Safety of Lycopene Oleoresin from Tomatoes

The applicant demonstrated the stability of the lycopene oleoresin from tomatoes under storage
at 4oC and room temperature for up to 37 months. Stability was assessed by spectrophotometry
and HPLC.

For the production of lycopene as a colour additive, the lycopene oleoresin from tomatoes
which is the subject of the present opinion is further processed by removal of part of the tomato
lipids to form the concentrated product, of which the final colour additive formulations are
prepared.
Attempts carried out including the Bradford assay for protein detection did not reveal residual
protein. The detection limit of the analysis for residual proteins was estimated to be about 1 ?g/g.
The identity and purity of lycopene oleoresin from tomatoes is in compliance with the purity
criteria for colouring agent E160d (lycopene) a food additive (EC, 1995).
II. Effect of the production process applied to the NF
The manufacturing process has been described by the applicant and is carried out in two steps:
the tomato pulp production and the extraction of the lycopene from the tomato pulp.

The production process for lycopene oleoresin from tomatoes is identical to that for the
production of the additive E160d, which has been examined and approved by the SCF (SCF,
1975, 1978), although E160d is prepared using an additional concentration step to obtain an
oleoresin that contains 60-70 % lycopene.

Lycopene oleoresin from tomatoes is produced by crushing lycopene-rich tomatoes to pulp. The
applicant states that the production process introduces no exogenous substances to the tomato pulp,
protects the tomato phytonutrients from oxidation and assures that the extraction is conducted on
unchanged and non-deteriorated raw material. The tomato pulp is then separated from the serum
and extracted with ethyl acetate. The extract is separated from the tomato pulp, and lycopene
oleoresin is obtained after solvent removal by evaporation under vacuum at 40-60o C.
Analytical methods to quantify the components of lycopene oleoresin and possible impurities
(organic volatiles, microbes, acid-soluble metals) were provided by the applicant.
Due to the chemical nature of lycopene (i.e. long chain with conjugated carbon-carbon double
bonds) it is susceptible to chemical changes such as isomerisation and degradation when exposed
to light, heat and oxygen (Lee and Chen, 2002). The applicant indicates that the production
process is designed such that it protects lycopene from oxidation.
The lycopene oleoresin obtained is packed under nitrogen and stored at 4 ºC.

III. History of the organism used as the source of the NF
High lycopene non-GMO tomatoes are used to produce the novel food. The applicant indicates
that conventional breeding methods utilising the natural gene pool of the genus Lycopersicon
have been applied in order to create a tomato plant with a high content of lycopene. This
particular variety is not consumed directly but is used in the production of tomato products.

The EFSA Journal (2008) 675, 9-22

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Safety of Lycopene Oleoresin from Tomatoes

IX. Anticipated intake/extent of use of the NF
The Panel notes that intake of lycopene may result from four sources including 1) normal
dietary intake from food, 2) intake from dietary supplements, 3) intake from proposed use in
food products and 4) use as a food colour.

Intake of lycopene from normal dietary sources
The applicant indicated that intakes of lycopene from natural sources may amount to a maximum
lycopene intake in the Netherlands (for the age group 55-69 years) from a normal diet by heavy
users of tomato products of 26.1 and 18.6 mg/day for men and women respectively,
corresponding to 0.44 mg/kg bw/day (assuming 60 kg bw for men) and 0.37 mg/kg bw/day
(assuming 50 kg bw for women).
An overview of average dietary exposure to lycopene from foods in different populations was
presented in previous EFSA evaluations (EFSA, 2005a and b; EFSA, 2008). It was concluded
that regular exposure to lycopene from natural dietary sources in different populations are,
according to dietary surveys, estimated to be on average between 0.5 and 5 mg/day, with high
exposures up to about 8 mg/day. High consumption of fruits and vegetables, especially tomato
products, may result in occasional exposure to 20 mg lycopene/day or more.

Intake of lycopene from proposed supplement use
The amount of lycopene used in dietary supplements is 5 to 15 mg/day, equivalent to 83-250 mg
lycopene oleoresin containing 6 % lycopene and smaller amounts when oleoresin with higher
lycopene content is used.

Intake of lycopene from use in food products at the proposed uses and use levels.
The applicant indicates that in case of addition to food, products will be formulated in such a way
that they will provide ca. 2 mg lycopene per serving. Table 2 presents an overview of the intended
use levels in the different food categories as provided by the applicant.

Table 2. Summary of recommended food uses and use levels of lycopene oleoresin in
different food categories proposed by the applicant.
Food Product
Added lycopene
Added lycopene
Food category
Portion Size (g)
(examples)
(mg/kg)
Per Serving (mg)
Yoghurt 16
125
2.0
Dairy products
Ice cream
25
80
2.0
Fruit and vegetable
Non-alcoholic
based beverages (not
8 250 2.0
flavoured drinks
as colorant)
Breakfast cereals
67
30
2.0
Cereal and cereal
products
Cereal bar
50
40
2.0
Soups (other than
Canned products
10 200 2.0
tomato)

The EFSA Journal (2008) 675, 10-22

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