EFSA Journal 2011; 9(5):2144
SCIENTIFIC OPINION
Scientific Opinion on the re-evaluation of lutein preparations other than
lutein with high concentrations of total saponified carotenoids at levels of at
least 80%1
EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS)2, 3
European Food Safety Authority (EFSA), Parma, Italy
ABSTRACT
Lutein is a carotenoid colour authorised as a food additive in the EU (E 161b) and reevaluated by the
Panel on Food Additives and Nutrient Sources added to Food (ANS) in 2010. The ANS Panel
established an ADI of 1 mg/kg bw/day and noted that this ADI refers to lutein derived from Tagetes
erecta containing at least 80% carotenoids. In the present opinion the Panel considered whether
additional studies made available address the gaps identified by the Panel in the toxicological database
for lutein preparations other than lutein with high concentrations of total saponified carotenoids at
levels of at least 80%. The Panel noted that in all additional studies made available, a specific lutein
ester preparation extracted from Tagetes erecta was tested, containing > 60% carotenoid esters (>93%
lutein esters, remainder zeaxanthin esters). No additional data were provided on lutein with levels of
~5-12% total carotenoids. The Panel concluded that lutein esters are not of concern with respect to
genotoxicity. The additional data also included a 90-day toxicity study and a reproductive and
developmental toxicity study. The Panel established for both studies a NOAEL of 1000 mg/kg bw/day,
the highest dose level tested (equivalent to 538 mg lutein equivalents/kg bw/day). The Panel noted that
this NOAEL of 538 mg lutein equivalents/kg bw/day is higher than the NOAEL of 200 mg/kg bw/day
(the highest dose level tested) in the 90-day rat study with lutein from which the ADI has been
derived. Based on these results, the Panel concluded that the additional database supports the
conclusion that the ADI of 1 mg/kg bw/day also refers to lutein with high concentrations of total
carotenoids extracted from Tagetes erecta and present as esters at levels of ≥ 60%. The Panel
concluded that the toxicological data-base available is too limited to conclude that the ADI also
applies to lutein preparations of lower purity or from other sources.
© European Food Safety Authority, 2011
1
2
On request of EFSA, Question No EFSA-Q-2010-01491, adopted on 13 April 2011.
Panel members: F. Aguilar, B. Dusemund, P. Galtier, J. Gilbert, D.M. Gott, S. Grilli, R. Gürtler, J. Koenig, C. Lambré, J-C.
Larsen, J-C. Leblanc, A. Mortensen, D. Parent-Massin, I. Pratt, I.M.C.M. Rietjens, I. Stankovic, P. Tobback, T. Verguieva,
R.A. Woutersen. Correspondence: ans@efsa.europa.eu
3
Acknowledgement: The Panel wishes to thank the members of the ANS Working Group B on Food Additives and Nutrient
Sources for the preparation of this opinion: D. Boskou, B. Dusemund, D. Gott, T. Hallas-Møller, A. Hearty, J. König, D.
Parent-Massin, I.M.C.M. Rietjens, G.J.A. Speijers, P. Tobback, T. Verguieva, R.A. Woutersen.
Suggested citation: EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS); Scientific Opinion on the reevaluation of lutein (E 161b) as a food additive. EFSA Journal 2011;9(5):2144. [25 pp.]. doi:10.2903/j.efsa.2011.2144.
Available online: www.efsa.europa.eu
© European Food Safety Authority, 2011
Re-evaluation of lutein (E 161b) as a food additive
KEY WORDS
Lutein, Lutein esters E 161b, CAS Registry Number 127-40-2, food colour, EINECS number 204-840-0.
SUMMARY
Lutein is a natural carotenoid colour authorised as a food additive in the EU (E 161b) and was
previously evaluated by the SCF in 1975 and JECFA in 2006 and re-evaluated by the Panel on Food
Additives and Nutrient Sources added to Food (ANS) in 2010.
JECFA established a group ADI of 0-2 mg/kg bw/day for lutein from Tagetes erecta and for
zeaxanthin. The SCF could not establish an ADI but concluded that xanthophylls prepared from
natural foods by physical processes are acceptable for use in food.
The ANS Panel established an ADI of 1 mg/kg bw/day, based on the NOAEL of 200 mg/kg bw/day
(the highest dose level tested) from a 90-day rat study, and an uncertainty factor of 200 due to the
absence of a multigeneration reproductive toxicity study and of chronic toxicity/carcinogenicity
studies.
The ANS Panel previously noted that this ADI refers to lutein derived from Tagetes erecta containing
at least 80% carotenoids consisting of lutein and zeaxanthin (79 and 5% respectively). In the previous
opinion re-evaluating the safety of lutein the ANS Panel concluded that the ADI of 1 mg/kg bw/day
does not refer to lutein preparations of lower purity or from other sources (EFSA, 2010).
The Panel previously also noted that other preparations of lutein are also on the market, (i.e. lutein
with ~5-12% total carotenoids, and lutein with ≥ 60 % total carotenoids present as esters). In the
previous evaluation the ANS Panel concluded that the toxicological data base available on these
preparations was too limited to conclude that the ADI also applies to these preparations.
In the present opinion the Panel considered whether the additional studies that were made available by
NATCOL might partially address the gaps identified by the Panel in the toxicological database for
lutein preparations other than lutein with high concentrations of total saponified carotenoids at levels
of at least 80%.
The Panel noted that a specific lutein ester preparation was tested in all additional studies made
available. This preparation was extracted from Tagetes erecta and contained > 60% carotenoid esters
(>93% lutein esters, remainder zeaxanthin esters). The remaining constituents were long-chain alkanes
and fatty acids with small amounts of terpenoids and triglycerides.
The Panel has been informed by NATCOL about the nature and levels of the terpenoids that may be
present. An essential oil extracted from Tagetes erecta flowers can contain a variety of terpenoids.
About 38% of the Tagetes erecta flower terpenes is D-limonene and 29.8% are cis- and trans-ocimene,
minor terpenes are linalool (5.9%), alpha-D-phellandrene (5.2%) and others. These figures are
confirmed by other researchers, who also identified piperitone, a constituent of mint leaves, and
caryophyllene, a constituent of cloves and basil, as important constituents. Minor amounts of these
terpenes may be present in the > 60% carotenoid esters extract. Being highly volatile substances they
are most likely removed during the manufacturing process together with the extraction solvents
hexane and iso-propanol. Since the limit value for extraction solvents in E161b is 50 mg/kg NATCOL
assumed that residual terpenes in the carotenoid esters extract do not exceed a low mg/kg level.
The Panel noted that terpenoids may be of toxicological relevance and that therefore specifications
may need to be updated to indicate the type and level of terpenoids present in the preparation.
The Panel has been informed by NATCOL that only hexane and iso-propanol (propan-2-ol) are used
as extraction solvents. The Panel noted that these solvents are already included in the specifications.
No additional data were provided on lutein with low concentrations of total carotenoids at levels of
~5-12%.
The additional studies provided included studies on bioavailability, demonstrating that lutein from
lutein esters is bioavailable, and that esterification does not impair lutein bioavailability in humans
EFSA Journal 2011;9(5):2144
2
Re-evaluation of lutein (E 161b) as a food additive
(Bowen et al., 2002). The Panel concluded that lutein from lutein esters is bioavailable to an extent
that is similar to the bioavailability of lutein itself.
The Panel noted that following passage through the gastrointestinal tract and/or uptake lutein esters are
hydrolysed to form free lutein again.
In the previous opinion on lutein the ANS Panel concluded that based on the studies available there
was no concern with respect to genotoxicity for lutein. Results from three genotoxicity studies using a
non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%) were made available by industry,
including a Salmonella typhimurium reverse mutation assay, a gene mutation assay in cultured
mammalian cells (L5178Y TK+/- mouse lymphoma cells) and an in vivo bone marrow micronucleus
test in the rat. No mutagenic activity was seen in any of the tests and the Panel concluded that lutein
esters are not of concern with respect to genotoxicity.
The additional data on the non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%) also
included a 90-day toxicity study performed according to OECD guideline 408 and under GLP, and a
reproductive and developmental toxicity study performed according to OECD 414 and under GLP. No
adverse effects were seen in the studies at any dose. The Panel established for both studies a NOAEL
of 1000 mg/kg bw/day, the highest dose level tested. Given that 10.23 mg lutein esters (nonhydrolysed lutein ester preparation from Tagetes erecta (> 60%)) equals 5.5 mg lutein equivalents, this
NOAEL would amount to 538 mg lutein equivalents/kg bw/day.
The Panel noted that this NOAEL of 538 mg lutein equivalents/kg bw/day is higher than the NOAEL
from which the ADI of 1 mg/kg bw/day has been derived. The absence of developmental toxicity at
dose levels up to 1000 mg/kg bw/day (the highest dose level tested) has also been reported for lutein
(from marigold extract; 79% lutein, 5% zeaxanthin).
Based on these results the Panel concluded that the additional database supports the extension of the
ADI of 1 mg/kg bw/day to lutein extracted from Tagetes erecta with ≥ 60 % total carotenoids present
as esters.
The Panel concluded that the ADI of 1 mg/kg bw/day refers to lutein derived from Tagetes erecta
containing ≥80% carotenoids consisting of lutein and zeaxanthin (79 and 5% respectively) and to
lutein with high concentrations of total carotenoids extracted from Tagetes erecta and present as esters
at levels of at least 60%.
The Panel concluded that the toxicological data-base available is too limited to conclude that the ADI
also applies to lutein preparations of lower purity or from other sources.
EFSA Journal 2011;9(5):2144
3
Re-evaluation of lutein (E 161b) as a food additive
TABLE OF CONTENTS
Abstract ....................................................................................................................................................... 1
Summary ..................................................................................................................................................... 2
Background as provided by the European Commission.............................................................................. 5
Terms of reference ...................................................................................................................................... 5
Assessment .................................................................................................................................................. 6
1.
Introduction ....................................................................................................................................... 6
2.
Technical data ................................................................................................................................... 6
2.1.
Identity of the substance ...................................................................................................... 6
2.2.
Specifications ...................................................................................................................... 8
2.3.
Manufacturing process ...................................................................................................... 10
2.4.
Methods of analysis in foods ............................................................................................. 10
2.5.
Reaction and fate in foods, stability .................................................................................. 10
2.6.
Case of need and proposed uses ........................................................................................ 10
2.7.
Information on existing authorisations and evaluations .................................................... 11
2.8.
Dietary exposure................................................................................................................ 12
3.
Biological and toxicological data .................................................................................................... 13
3.1.
Absorption, distribution, metabolism and excretion ......................................................... 14
3.2.
Toxicological data ............................................................................................................. 15
3.2.1.
Acute oral toxicity ................................................................................................................ 15
3.2.2.
Short-term and subchronic toxicity....................................................................................... 15
3.2.3.
Genotoxicity ......................................................................................................................... 16
3.2.4.
Chronic toxicity and carcinogenicity .................................................................................... 17
3.2.5.
Reproductive and developmental toxicity ............................................................................ 17
3.2.6.
Human data ........................................................................................................................... 18
4.
Discussion ....................................................................................................................................... 18
Conclusions ............................................................................................................................................... 20
Documentation provided to EFSA ............................................................................................................ 21
References ................................................................................................................................................. 21
Glossary/Abbreviations ............................................................................................................................. 24
EFSA Journal 2011;9(5):2144
4
Re-evaluation of lutein (E 161b) as a food additive
BACKGROUND AS PROVIDED BY THE EUROPEAN COMMISSION
According to the framework Directive 89/107/EEC4 on food additives, the Scientific Committee for
Food (SCF) should be consulted before the adoption of provisions likely to affect public health, such
as the drawing up of lists of additives and the conditions for their use. Accordingly, all food additives,
prior to their authorization, have been evaluated for their safety by the SCF or by its successor the
European Food Safety Authority (EFSA).
Directive 89/107/EEC as well as Regulation (EC) No 1333/2008 of the European Parliament and of
the Council of 16 December 2008 on food additives5 which will apply as from 20 January 2010,
require that food additives must be kept under continuous observation and must be re-evaluated
whenever necessary in the light of changing conditions of use and new scientific information. In
addition Regulation (EC) No 1333/2008 requires that all food additives which were permitted before
20 January 2009 shall be subject to a new risk assessment carried out by EFSA. Regulation (EC) No
257/2010 of 25 March 2010 sets up a programme for the re-evaluation of approved food additives in
accordance with Regulation (EC) No 1333/2008.
On 7 July 2010, the Scientific Panel on Food Additives and Nutrient Sources added to food (ANS)
adopted a scientific opinion on the re-evaluation of lutein (E 161b) as a food additive.
On 30 August 2010 NATCOL and CEFIC sent a letter to EFSA related to the opinion of the ANS
Panel on lutein. In this letter, several issues were raised including the fact that data that had not been
considered by the Panel could address some of the data gaps identified in the opinion.
During its eighteenth plenary meeting in October 2010, the ANS Panel has discussed the issues raised
in the letter by NATCOL and CEFIC. The Panel has considered that the studies on lutein esters that
can be made available by NATCOL might partially address the gaps identified by the Panel in the
toxicological database for lutein preparations other than lutein with high concentrations of total
saponified carotenoids at levels of at least 80%. Therefore the Panel proposes to EFSA a self-task to
re-evaluate the safety of these other lutein preparations.
TERMS OF REFERENCE
The European Food Safety Authority asks its Scientific Panel on Food Additives and Nutrient Sources
added to food to re-evaluate the safety as a food additive of lutein preparations complying with the
purity criteria defined in the Directive EC No 2008/128 of 22 December 2008 and that do not contain
high concentrations of total saponified carotenoids at levels of at least 80%.
4
5
OJ L 40, 11.2.1989, p. 27
OJ L 354, 31.12.2008, p. 16.
EFSA Journal 2011;9(5):2144
5
Re-evaluation of lutein (E 161b) as a food additive
ASSESSMENT
1.
Introduction
Lutein is a natural carotenoid colour authorised as a food additive in the EU (E 161b) and was
previously re-evaluated by the Panel on Food Additives and Nutrient Sources added to Food
(ANS)(EFSA, 2010).
The Panel established an Acceptable Daily Intake (ADI) of 1 mg/kg bw/day, based on a No-ObservedAdverse-Effect Level (NOAEL) of 200 mg/kg bw/day which was the highest dose level tested) in a
90-day rat study. An uncertainty factor of 200 was applied because of the absence of a multigeneration reproductive toxicity study and of chronic toxicity/carcinogenicity studies.
The Panel noted that this ADI refers to lutein derived from Tagetes erecta containing at least 80%
carotenoids consisting of lutein and zeaxanthin (79 and 5% respectively). According to specifications
provided by the Natural Food Colours Association (NATCOL) this may refer to the lutein with high
concentrations of total saponified carotenoids at levels of at least 80% (cf. JECFA specifications for
lutein from Tagetes erecta). The Panel concluded that the ADI does not refer to lutein preparations of
lower purity or from other sources.
The Panel also noted that other preparations of lutein are also on the market, i.e. lutein with low
concentrations of total carotenoids at levels of ~5-12%, and lutein with high concentrations of total
carotenoids extracted and present as esters at levels of at least 60%. The Panel concluded that the
toxicological data base available on these preparations was too limited to conclude that the ADI also
applies to these preparations.
The Panel has considered that the studies on lutein esters that were made available by NATCOL might
partially address the gaps previously identified by the Panel in the toxicological database for lutein
preparations other than lutein with high concentrations of total saponified carotenoids at levels of at
least 80%.
The Panel noted that a chemically more correct name for total saponified carotenoids would be total
saponified xanthophyll esters, so in what follows the term total saponified carotenoids has to be
interpreted as such.
Thus, the present opinion deals with the re-evaluation of the safety as a food additive of lutein
preparations that comply with the purity criteria defined in the Directive 2008/1286 and that do not
contain high concentrations of total saponified carotenoids at levels of at least 80%.
2.
Technical data
2.1.
Identity of the substance
Lutein (E 161b) is a natural carotenoid colour with the formula C40H56O2 and a molecular weight of
568.88 g/mol. Its chemical name has been described as 4’,5’-didehydro-5’,6’-dihydro-,-carotene3,3’-diol. The CAS number is 127-40-2 and the EINECS number is 204-840-0.
The structural formula of lutein is given in Figure 1:
H3C
HO
6
CH3
CH3
CH3
H3C
CH3
CH3
CH3
H3C
OH
CH3
Commission Directive 2008/128/EC of 22 December 2008 laying down specific purity criteria concerning colours for use in
foodstuffs. OJ L 6, 10.1.2009, p. 2
EFSA Journal 2011;9(5):2144
6
Re-evaluation of lutein (E 161b) as a food additive
Figure 1:
Structural formula of lutein
The Panel has been informed by NATCOL that the following products are marketed under the current
EU specifications:
a)
Lutein with low concentrations of total carotenoids at levels of ~5-12%.
b)
Lutein with high concentrations of total carotenoids extracted and present as esters at levels of
at least 60%, extracted from Tagetes erecta.
c)
Lutein with high concentrations of total saponified carotenoids at levels of at least 80% (cf.
JECFA specifications for lutein from Tagetes erecta).
The present opinion deals especially with the first two of these lutein preparations since it re-evaluates
lutein preparations other than lutein with high concentrations of total saponified carotenoids at levels
of at least 80%.
The Panel has been informed by NATCOL about the state of the different preparations. The raw
extract of lutein with high concentrations of total carotenoids is specified to contain > 60% carotenoid
esters. Typically, the raw carotenoid esters extract contains > 80% carotenoids esters out of which at
least 93% are lutein esters. The rest is other carotenoids such as zeaxanthin esters. This high
concentrate extract is a solid. Subsequently, vegetable oil is added to adjust the product to 15% to 37%
in carotenoid esters (equal to 14-34% lutein esters), which are the commercial products. These
standardised lutein ester products are liquids. The lutein ester concentrate is not marketed as such but
only in the standardised form diluted with vegetable oil.
In second instance, NATCOL informed the Panel that to their knowledge there are no lutein ester
products on the market with concentrations of total carotenoids at levels of ~5-12%. Concentrations
for commercial products range between 15-37% (as carotenoid esters which corresponds to 14-34%
lutein esters). There is one product on the market at 4% carotenoid esters (which corresponds to 3.7%
lutein esters) which is formulated using a solid carrier. This product is a solid.
EFSA Journal 2011;9(5):2144
7
Re-evaluation of lutein (E 161b) as a food additive
2.2.
Specifications
Specifications for lutein (E 161b) have been defined in the EU legislation (Directive 2008/128/EC)
and by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) (JECFA, 2006b) (Table
1) and were described and discussed in detail in the scientific opinion on the re-evaluation of lutein (E
161b) as a food additive (EFSA, 2010).
Table 1:
Specifications for lutein (E 161b) according to Commission Directive 2008/128/EC and
for mixed carotenoids, Tagets extract (INS No. 161b(ii)) and lutein from Tagetes erecta
(INS No. 161b(i)) according to JECFA (JECFA, 2006)
Commission
Directive
2008/128/EC
(Lutein)
Assay
Description
Solvent residues
Acetone
Methanol
Ethanol
Propan-2-ol
Hexane
Methyl ethyl
ketone
Dichloromethane
Propylene glycol
Moisture
Ash
Zeaxanthin
Synthetic colours
Waxes
Arsenic
Lead
Mercury
Cadmium
Heavy metals (as
Pb)
Content of total
colouring matter ≤ 4
% calculated as lutein
Dark, yellowish
brown liquid
≤ 50 mg/kg, singly or
in combination
JECFA (2006)
(Lutein from
Tagetes erecta)
JECFA (2006)
(Mixed carotenoids)
JECFA (2006)
(Tagetes extract)
Total colouring matter (as
lutein) not less than
declared
Total colouring
matter (as lutein)
not less than
declared
Dark yellowbrown liquid
≤ 80% total
carotenoids,
≤ 70% lutein
≤ 50 mg/kg
-
≤ 10 mg/kg
≤ 50 mg/kg
-
-
≤ 1000 mg/kg
≤ 1.0 %
≤ 1.0 %
≤ 9.0 %
-
≤ 2 mg/kg
-
≤ 14.0 %
≤ 3 mg/kg
-
Dark, yellowish brown
liquids with a weak haylike odour
≤ 50 mg/kg, singly or in
combination
A free-flowing,
orange-red
powder
≤ 10mg/kg, individually
≤ 10 mg/kg
-
≤ 3 mg/kg
≤ 10 mg/kg
≤ 1 mg/kg
≤ 1 mg/kg
≤ 40mg/kg
Free of synthetic
pigments, canthaxanthin,
and apocarotenoic acid
ethyl ester
≤ 5 mg/kg
-
The Panel noted that neither the EU nor the JECFA specifications provide information on the % of
subsidiary colouring matter.
The Panel has been informed by NATCOL that three products are marketed under the current EU
specifications (see section 2.1).
EFSA Journal 2011;9(5):2144
8
Re-evaluation of lutein (E 161b) as a food additive
The present opinion deals especially with the first two of these lutein preparations since it re-evaluates
lutein preparations other than lutein with high concentrations of total saponified carotenoids at levels
of at least 80%.
Lutein with low concentrations of total carotenoids at levels of ~5-12%
The pigment contribution of products on the market is not less than 4% calculated as lutein. The main
colouring principle of these preparations of lutein consists of carotenoids of which lutein and its fatty
acid esters account for the major part. Variable amounts of carotenes will also be present. The product
may contain fats, oils and waxes naturally occurring in the source material. It is insoluble in water and
soluble in hexane.
The product complies in all aspects with the purity criteria laid down in Directive 2008/128/EC.
According to NATCOL this colour is not used for aluminium lakes.
Lutein with high concentrations of total carotenoids extracted and present as esters at levels of at
least 60%
The Panel noted that the new data provided by industry deal with a specific lutein ester preparation.
The chemical identify of this lutein ester preparation is described as follows:
The carotenoid composition of this product reflects the carotenoid pattern naturally occurring in
Tagetes erecta and comprises primarily lutein diesters and lutein monoesters. The non-standardised
Tagetes extract contains > 60% in carotenoid esters (>93% lutein esters, rest zeaxanthin esters), the
remaining constituents being long-chain alkanes, fatty acids with small amounts of terpenoids and
triglycerides. The “high concentrate” extract is sold as such or standardised by means of food grade
vegetable oils (e.g. soybean).
The Panel has been informed by NATCOL about the nature and levels of the terpenoids that may be
present. An essential oil extracted from Tagetes erecta flowers can contain a variety of terpenoids
(Hagers Handbuch der pharmazeutischen Praxis, 1979). About 38% of the Tagetes erecta flower
terpenes is D-limonene and 29.8% are cis- and trans-ocimene, minor terpenes are linalool (5.9%),
alpha-D-phellandrene (5.2 %) and others. These figures are confirmed by other researchers, who also
identified piperitone, a constituent of mint leaves, and caryophyllene, a constituent of cloves and basil,
as important constituents (El-Tantawy et al., 1994; Marotti et al., 2004). Minor amounts of these
terpenes may be present in the > 60% carotenoid esters extract. Being highly volatile substances they
are most likely removed during the manufacturing process together with the extraction solvents
hexane and iso-propanol. Since the limit value for extraction solvents in E161b is 50 mg/kg NATCOL
assumed that residual terpenes in the carotenoid esters extract do not exceed a low mg/kg level.
The Panel noted that terpenoids may be of toxicological relevance and that therefore specifications
may need to be updated to indicate the type and level of terpenoids present in the preparation.
The Panel has been informed by NATCOL that only hexane and iso-propanol (propan-2-ol) are used
as extraction solvents. The Panel noted that these solvents are already included in the specifications.
The product complies in all aspects with the purity criteria laid down in Directive 2008/128/EC.
Aromatic amines do not occur in this colour, and it is not used for aluminium lakes.
Upon request industry further informed that this specific lutein ester preparation contains between 0.5
and 1% free lutein. The preparation is extracted from Tagetes erecta and it is not hydrolysed unlike it
is the case with other lutein sources that have a higher concentration of free lutein.
The Panel noted that the EU specifications for lutein E 161 b states that “The main colouring principle
consists of carotenoids of which lutein and its fatty acid esters account for the major part.”, whereas
in the JEFCA specifications lutein esters are not mentioned. The JECFA specifications mention
saponification which would imply that esters are hydrolysed.
EFSA Journal 2011;9(5):2144
9
Re-evaluation of lutein (E 161b) as a food additive
2.3.
Manufacturing process
The manufacturing processes for lutein (E 161b) have been described in the scientific opinion on the
re-evaluation of lutein (E 161b) as a food additive (EFSA, 2010).
The present opinion only presents additional data made available by NATCOL which might partially
address the gaps identified by the Panel in the toxicological database for lutein preparations other than
lutein with high concentrations of total saponified carotenoids at levels of at least 80%.
Lutein with low concentrations of total carotenoids at levels of ~5-12
This lutein preparation is obtained by solvent extraction of the natural strains of edible fruits and
plants, grass, Lucerne (alfalfa) and Tagetes erecta. Only the following solvents are used for extraction:
methanol, ethanol, propan-2-ol, hexane, acetone, methyl ethyl ketone, dichloromethane and carbon
dioxide.
Lutein with high concentrations of total carotenoids extracted and present as esters at levels of at
least 60%
The Panel noted that the additional toxicological data provided by industry deal especially with a
specific lutein ester preparation that is > 60% in carotenoid esters (>93% lutein esters, rest zeaxanthin
esters), the remaining constituents being long-chain alkanes, fatty acids with small amounts of
terpenoids and triglycerides.
The manufacturing procedure for this lutein ester preparation is as follows:
Marigold flowers (Tagetes erecta) are used as the commercial source of lutein esters. The marigold
flowers used for production of the lutein ester preparation are grown under controlled conditions on
the premise of contract farmers. Food grade solvents approved for the manufacture of food are used in
the extraction steps. The aim of the initial processing steps is to extract the carotenoid esters from the
marigold flowers and to purify the resulting extract from other naturally occurring plant cell
constituents. This is achieved by proprietary physical separation steps involving drying, milling,
solvent extraction (hexane and iso-propanol), purification and concentration. The resulting product is a
natural carotenoid ester extract.
2.4.
Methods of analysis in foods
The methods of analysis in foods for lutein (E 161b) have been described in the scientific opinion on
the re-evaluation of lutein (E 161b) as a food additive (EFSA, 2010).
2.5.
Reaction and fate in foods, stability
The stability of lutein has been assessed in two liquid enteral nutrition products (milk-based). The
lutein content in these products was analysed every three months, over a 12-month period and results
showed that the levels remained within ± 10% of the original value over this period. As lutein is
sensitive to oxygen and light, it should be stored in sealed containers and in the dark (EFSA, 2006).
2.6.
Case of need and proposed uses
Lutein is currently an authorised natural food colour in the EU, with maximal allowed use levels of 50
to 500 mg/kg food for various foodstuffs.
Details on the case of need and proposed uses for lutein (E 161b) have been described in the scientific
opinion on the re-evaluation of lutein (E 161b) as a food additive (EFSA, 2010).
EFSA Journal 2011;9(5):2144
10
Re-evaluation of lutein (E 161b) as a food additive
2.7.
Information on existing authorisations and evaluations
Lutein has been previously evaluated by the EU Scientific Committee for Food (SCF) in 1975 and
JECFA in 2004 and 2006, and re-evaluated by the European Food Safety Authority (EFSA) in 2010.
The SCF indicated that no specific biological data were available (1975) and therefore could not
establish an ADI, but nevertheless recommended that xanthophylls prepared from natural foods by
physical processes be accepted for use as colouring matters in food without further investigation. For
the purposes of the Council Directive 62/2645/EEC7, the SCF suggested that the acceptable natural
xanthophylls be defined as including the 3-hydroxy- and 3,3-dihydroxy-neoxanthin, neochrome, and
the fatty acid esters of these compounds present in natural foods.
In 1977, the SCF concluded that no ADI could be established for antheraxanthin oleoresin (hexane
extract of the flower petals of Aztec Marigold (=Tagetes erecta)) and that antheraxanthin oleoresin is
not toxicologically acceptable for use in food.
JECFA established a group ADI of 0-2 mg/kg body weight (bw) for lutein from Tagetes erecta and for
zeaxanthin (JECFA, 2004; 2006). The ADI was derived from a 90-day study in rats (Kruger et al.,
2002; Pfannkuch et al., 2000; Pfannkuch et al., 2001). The ADI was based on the NOAEL of this
study of 200 mg/kg bw/day (the highest dose tested) and an uncertainty factor of 100. Although the
ADI was based on the results of a short-term study, JECFA took account of supporting data and lack
of effects at much higher doses in some other studies (e.g. a study of developmental toxicity) in
concluding that an uncertainty factor of 100 was appropriate. Zeaxanthin was included in the ADI due
to the toxicological data and structural and physiological similarities between the xanthophylls lutein
and zeaxanthin. The group ADI does not apply to other xanthophyll-containing extracts with a lutein
or zeaxanthin content lower than that cited in the specifications. According to the specifications of
JECFA (JECFA, 2006b), lutein from Tagetes erecta should contain ‘not less than 80% total
carotenoids, not less than 70% lutein’. However, according to the ‘mixed carotenoids’ specifications
of JECFA (2006b), total colouring matter (as lutein) should be ‘not less than declared’. The Panel
noted that it is not completely clear how the JECFA group ADI relates to these ‘mixed carotenoids’
specifications.
The EFSA Panel on Additives, Flavourings, Processing Aids and Materials in Contact with Food
(AFC) evaluated lutein for use in the manufacture of Foods prepared for Particular Nutritional Uses
(PARNUTs)(EFSA, 2006). The product evaluated had the same specifications as the approved food
additive. The petitioner’s proposed use of lutein was for Foods for Special Medical Purposes (FSMPs)
at levels that would give rise to daily intakes of 0.5–2 mg lutein/day. This is within the range of a
regular dietary intake. Given that lutein extracted from the natural strains of edible fruits and plants,
grass, Lucerne and Tagetes erecta is already permitted as a food additive, the AFC Panel concluded
that the use of lutein in FSMPs is not of safety concern under the proposed use levels which are in the
range of the regular dietary intake of lutein, provided that the preparation is in compliance with the
existing EU specifications for the food additive. The Panel was not able to evaluate the general use of
lutein in PARNUTS, since no information was provided on proposed uses and use levels other than for
FSMPs (EFSA, 2006).
The EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) issued an opinion on the safety,
bioavailability and suitability of lutein for the particular nutritional use by infants and young children
(EFSA, 2008b). The Panel considered that the information provided in the dossier did not raise
concerns about the safety of lutein in infant formulae at the levels achieved through the natural content
of ingredients nor at the level of use (concentration of added lutein 250μg/l) proposed by the applicant
for infant formulae with a low natural lutein content (about 20 μg/l or lower).
The EFSA Panel on Additives and Products or Substances use in Animal Feed (FEEDAP) issued an
opinion on the safety of use of colouring agents, including lutein, in animal nutrition (EFSA, 2009). It
7
Council Directive 62/2645/EECof 23 October 1962 on the approximation of the rules of the Member States concerning the
colouring matters authorized for use in foodstuffs intended for human consumption. OJ 115, 11.11.1962, p. 2645–2654.
EFSA Journal 2011;9(5):2144
11
Re-evaluation of lutein (E 161b) as a food additive
was stated that data on the safety of lutein for the target animals are not available, but that given the
widespread natural occurrence of the compounds and considering the molecular structure of the
xanthophylls, the FEEDAP Panel does not see any reason for concern. It was also concluded that
taking into account the human lutein and zeaxanthin intake from all sources, the contribution from
food of animal origin (eggs and poultry tissues produced with lutein- and zeaxanthin containing diets)
would be a very small proportion of the total intake which varies with the consumption pattern in
different countries, and that it does not require a particular safety assessment.
The EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS) re-evaluated the
safety of lutein (E 161b) as a food additive (EFSA, 2010). The Panel established an ADI of 1 mg/kg
bw/day, based on the NOAEL of 200 mg/kg bw/day (the highest dose level tested) from a 90-day rat
study, and an uncertainty factor of 200, applied to compensate for the absence of a multigeneration
reproductive toxicity study and of chronic toxicity/carcinogenicity studies.
The ANS Panel noted that this ADI refers to lutein derived from Tagetes erecta containing ≥ 80%
carotenoids consisting of lutein and zeaxanthin (79 and 5% respectively). According to specifications
provided by NATCOL this may refer to the lutein with high concentrations of total saponified
carotenoids at levels of ≥ 80% (cf. JECFA specifications for lutein from Tagetes erecta). The Panel
concluded that the ADI does not refer to lutein preparations of lower purity or from other sources.
The ANS Panel also noted that other preparations of lutein are also on the market, i.e. lutein with low
concentrations of total carotenoids at levels of ~5-12%, and lutein with high concentrations of total
carotenoids extracted and present as esters at levels of ≥ 60%. The ANS Panel concluded that the
toxicological data base available on these preparations was too limited to conclude that the ADI also
applied to these preparations.
2.8.
Dietary exposure
Dietary exposure estimates for lutein (E 161b) have been described in the scientific opinion on the reevaluation of lutein (E 161b) as a food additive (EFSA, 2010).
Table 2 summarises the anticipated exposure of children and adults to lutein, as calculated in the
previous EFSA opinion (EFSA, 2010).
Table 2:
Summary of anticipated exposure to lutein using tiered approach (EC, 2001) in children
and adult populations
Adult UK
population
(>18 years old)
Tier 1. Budget method
Tier 2. Maximum Permitted Level
 Mean exposure
 Exposure 95th *or 97.5th percentile
Tier 3. Maximum reported use levels
 Mean exposure
 Exposure 95th *or 97.5th percentile
8.1
Pre-school UK
children
(1.5 - 4.5 years old,
15 kg body weight)
mg/kg bw/day
13.1
Children EXPOCHI
population
(1-10 years old,
25-30 kg body weight)
0.8
3.2
3.0
7.2
0.5-3.4
1.2-7.2
0.6
1.6
2.2
5.7
0.2-2.2
0.7-5.7
* For EU children, estimates are based on the EXPOCHI report, which gives the 95th percentile intake.
** For UK, estimates are based on the UNESDA report which gives the 97.5th percentile intake from beverages
plus per capita average from the rest of diet (Tennant, 2006).
The ANS Panel stated the following (EFSA, 2010):
EFSA Journal 2011;9(5):2144
12
Re-evaluation of lutein (E 161b) as a food additive
“In the case of lutein, when considering MPLs of use (Tier 2), the mean dietary exposure of UK
children aged 1.5 to 4.5 years and weighing 15 kg, was 3.0 mg/kg bw/day and 7.2 mg/kg bw/day for
high level (97.5th percentile) consumers of soft drinks. The mean dietary exposure of European
children (aged 1-10 years and weighing 16-29 kg) considered by the EXPOCHI consortium ranged
from 0.5 to 3.4 mg/kg bw/day, and from 1.2 to 7.2 mg/kg bw/day at the 95th percentile. Estimates
reported for the UK adult population give a mean dietary exposure to lutein of 0.8 mg/kg bw/day, and
of 2.0 mg/kg bw/day for high level (97.5th percentile) consumers of soft drinks.
[…]
Tier 3 intake estimates, based on the maximum use levels from the NATCOL usage survey, ranged
from 0.6-2.2 mg/kg bw/day. High level intakes ranged from 0.7-5.7 mg/kg bw/day. Therefore, at the
current use levels, the ADI of 1 mg/kg bw/day will be exceeded due to the use of lutein as a food
colour at the upper end of the range. Furthermore, EFSA (2006) indicated that overall the dietary
intake of lutein as such is estimated to be between 0.8 and 2.5 mg/day, equivalent to 0.01 – 0.04 mg/kg
bw/day for a 60 kg person, indicating that the worst case scenario for intake of lutein used as a food
colour in combination with its average intake from other dietary sources does exceed the ADI of 1
mg/kg bw/day.
[…]
The Panel concluded that at the current levels of use Tier 3 intake estimates are above the ADI of 1
mg/kg bw/day at the upper end of the range.
The Panel concluded that the average intake for adults from the regular diet amounts to 1-4 % of the
ADI of 1 mg/kg bw/day. High level intakes from the regular diet would amount to 28% of this ADI for
children (assuming an intake of lutein present in food of 7 mg/day and a body weight of 25 kg, equal
to 0.28 mg/kg bw/day).”
The Panel recently received from industry corrected use levels to replace those provided previously
that were not related to the content of pigment. This change is anticipated to have an important impact
on Tier-3 exposure estimates. Therefore, the Panel will propose a self-task mandate for the reevaluation of the exposure estimates and their comparison to the ADI.
3.
Biological and toxicological data
Lutein has been previously evaluated by the SCF in 1975, by JECFA in 2006, TemaNord in 2002 and
EFSA in 2010. The present opinion presents only additional data made available by NATCOL which
might partially address the gaps identified by the Panel in the toxicological database for lutein
preparations other than lutein with high concentrations of total saponified carotenoids at levels of at
least 80%.
The studies described in this section were performed mostly between 1990 and 2008. Several studies
complied with OECD guidelines and/or Good Laboratory Practice (GLP) principles but for others
compliance is unclear.
The Panel noted that in all additional studies made available and now presented the preparation tested
was a specific lutein ester preparation that is > 60% in carotenoid esters (>93% lutein, rest
zeaxanthin), the remaining constituents being long-chain alkanes, fatty acids with small amounts of
terpenoids and triglycerides. This preparation will be further referred to as “a non-hydrolysed lutein
ester preparation from Tagetes erecta (> 60%)”.
EFSA Journal 2011;9(5):2144
13
Re-evaluation of lutein (E 161b) as a food additive
3.1.
Absorption, distribution, metabolism and excretion
The Panel noted that there are no studies in experimental animals on bioavailability of lutein esters
available.
Human studies
Subjects received 10 mg lutein derived from marigold daily during 12 weeks. Mean plasma lutein
concentration increased from 0.18 to 0.90 μmol/l within the first four weeks and stayed at this level
during the supplementation period (Berendschot et al., 2000). The Panel was informed that the lutein
used in this study was a non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%).
Bowen et al. (2002) reported that esterification does not impair lutein bioavailability in humans. In this
study lutein bioavailability was determined for lutein diester and unesterified lutein formulations.
Healthy subjects (n = 18) consumed a single dose of each formulation (either 0.5 or 0.67 μmol
lutein/kg body, 10 and 8 subjects, respectively) in random order, and the appearance of free lutein plus
zeaxanthin was measured in serum from 0 to 408 hours. As a measure of bioavailability, areas under
the serum concentration x time curves (AUC) were independent of gender, body mass index and lutein
dose. The mean AUC for the subjects consuming the lutein diester formulation amounted to 37.0 +
35.8 nmol/L.h compared to 22.9 + 19.0 nmol/L.h for the subjects consuming the lutein formulation.
Comparison with data from previous studies suggested that dissolution was a greater limitation to
bioavailability than lutein ester hydrolysis because an oil-solubilised unesterified lutein preparation,
given at 0.5 μmol/kg body, resulted in greater mean peak concentrations and AUC compared with
either the unesterified or lutein diester formulations used in the study, because in this case the AUC
amounted to 59.6 + 25.5 nmol/L.h. The authors concluded that the lutein diester formulation poses no
impediment to lutein bioavailability at the doses tested, but that formulation dissolution is an
important factor in lutein bioavailability.
Bone et al. (2003) reported that lutein dietary supplements, at dose levels ranging from 2.4 to 30
mg/day, raise macular pigment density and serum concentrations of this carotenoid in humans. Serum
lutein concentrations in each subject reached a plateau that was correlated with the dose (r = 0.82, P <
0.001). Plateau concentrations ranged from 2.8 x 10-7 to 2.7 x 10-6 mol/l. The rate of increase in
macular pigment optical density was correlated with the plateau concentration of carotenoid in the
serum (r = 0.58, P < 0.001). The Panel was informed that the lutein used in one of the experiments in
this study was a non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%).
Chung et al. (2004) reported on the bioavailability of various forms of lutein including lutein esters. In
this study healthy men (n = 10) participated in an intervention study with a crossover design. After a
2-week washout period during which they consumed a low-carotenoid diet, the men were administered
1 of 4 lutein doses (lutein supplement, lutein ester supplement, spinach, and lutein-enriched egg) for 9
days. All lutein doses provided 6 mg lutein except for the lutein ester dose, which provided 5.5 mg
lutein equivalents. Serum samples were collected from fasting subjects on day –14, 1 (baseline), 2, 3,
and 10 and analysed for lutein concentration. Triacylglycerol-rich lipoproteins (TRL) were separated
from postprandial blood samples (0–24 hours) after the first lutein dose and analyzed for lutein
concentration. Subjects completed all 4 treatments of the study in random order. Results from
repeated-measures 1-way ANOVA results showed that the baseline and dose-adjusted lutein response
in serum was significantly higher after egg consumption than after lutein, lutein ester, and spinach
consumption on day 10. There was no significant difference in TRL response. Koh et al. (2004)
evaluated macular pigment optical density (MPOD) and plasma concentrations of lutein in patients
with early age-related maculopathy (ARM) following a daily 20 mg lutein ester (non-hydrolysed
lutein ester preparation from Tagetes erecta (> 60%)(equivalent of 10 mg/day free lutein) supplement.
The preparation tested was non-hydrolysed lutein ester from Tagetes erecta (> 60%) which provided a
dose equivalent to 10 mg/day free lutein. The period of lutein supplementation ranged from 18 to 21
weeks. MPOD measurements and blood samples (without fasting) were taken every two to six weeks
during the period of lutein supplementation and at least once (2–9 weeks) after the cessation of
EFSA Journal 2011;9(5):2144
14
Re-evaluation of lutein (E 161b) as a food additive
supplement. Plasma lutein increased from a mean (SD) baseline concentration of 182 (127) ng/ml to a
peak of 1077 (165) ng/ml in ARM patients, and from 152 (57) to 1110 (605) ng/ml in control subjects.
Trieschmann et al. (2007) investigated the response to supplemental lutein and zeaxanthin and coantioxidants; they measured macular pigment optical density (MPOD) and serum concentrations of its
constituent carotenoids, response to supplemental lutein and zeaxanthin and co-antioxidants. An
intervention (I) group, consisting of 108 subjects (age 71.5 ±7.1] years), of which 92.6% exhibited
features of age-related macular degeneration (AMD), received a daily supplement consisting of 12 mg
lutein and 1 mg zeaxanthin, 120 mg vitamin C, 17.6 mg vitamin E, 10 mg zinc, 40 mg selenium for a
period of 6 months. The xanthophyll supplements were both provided as esters, the lutein esters being
a non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%). A control group of 28 subjects
(age 71.0 ± 8.1 years), who received no dietary supplementation or modification, was examined at
baseline and once again after a mean of 29.4 (±9.3) weeks. In spite of rises in serum concentrations of
lutein and zeaxanthin in the intervention group, there remained a substantial proportion of subjects for
whom MPOD augmentation in response to supplemental lutein, zeaxanthin and co-antioxidants could
not be detected over the study period. This indicated that intestinal malabsorption of these carotenoids
is not responsible for the lack of a macular response to such supplements.
The Panel concluded that lutein from lutein esters is bioavailable to an extent that is similar to the
bioavailability of lutein itself.
3.2.
Toxicological data
3.2.1.
Acute oral toxicity
The Panel previously concluded that the acute oral toxicity of lutein is low (EFSA, 2010).
Data from a 14-day acute oral toxicity rest in rats (10 controls and 10 exposed) on lutein diester
oleoresin (non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%)) were submitted to
EFSA (CLG 1998a). The study was conducted in compliance with GLP and the lutein preparation
showed no evidence of toxicity at a dose of 3.75 g/kg bw in rats.
3.2.2.
Short-term and subchronic toxicity
A 90-day repeated dose toxicity study in rats with a non-hydrolysed lutein ester preparation from
Tagetes erecta (> 60%) was made available for evaluation (Wierich and Leuschner, 2006a). The study
was performed according to OECD guideline 408 and in compliance with GLP. Groups of 10 rats of
each sex were given 0 (vehicle control), 100, 300 or 1000 mg non-hydrolysed lutein ester preparation
from Tagetes erecta (> 60%)/kg bw/day by gavage for 90 days. Additionally 5 rats of each sex were
given either 0 or 1000 mg doses by gavage; after 90 days of exposure these animals were kept for a
recovery period of 6 weeks. No treatment-related clinical findings, ophthalmoscopy findings,
functional observations or mortality were observed at any dose. Body weights, food and water intake
were not influenced by treatment in any group. Haematology, clinical chemistry urinalysis and organ
weights were unaffected by treatment. No treatment-related changes were seen in organs/tissues either
macroscopically or microscopically or in the oestrus cycle of females or numbers of spermatozoa and
morphology of spermatids of males at any dose.
The study authors concluded that the NOAEL of this study was 1000 mg non-hydrolysed lutein ester
preparation from Tagetes erecta (> 60%)/kg bw/day, the highest dose tested. The Panel agreed with
this NOAEL.
Given that 10.23 mg lutein esters (non-hydrolysed lutein ester preparation from Tagetes erecta (>
60%)) equals 5.5 mg lutein equivalents (Chung et al., 2004), this NOAEL would amount to 538 mg
lutein equivalents/kg bw/day. This conversion factor is based on the molecular weight of lutein of
568.88 g/mol, the lutein esters being diesters and an average molecular weight of a fatty acid assumed
to be about 245 g/mol.
EFSA Journal 2011;9(5):2144
15
Re-evaluation of lutein (E 161b) as a food additive
3.2.3.
Genotoxicity
In the previous opinion on lutein the ANS Panel concluded that based on the studies available there is
no concern with respect to genotoxicity for lutein (EFSA, 2010).
Results from three new genotoxicity studies of a non-hydrolysed lutein ester preparation from Tagetes
erecta (> 60%) were made available by industry, including an Ames test, an in vitro mammalian
genotoxicity mouse lymphoma assay and an in vivo micronucleus test, and these studies were
evaluated in this opinion
The non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%) was tested in the Salmonella
typhimurium strains TA98, TA100, TA102, TA1535 and TA1537 according to OECD guideline 471
and in compliance with GLP, in two independent experiments with and without metabolic activation
(a microsomal preparation derived from Aroclor 1254-induced rat liver) (Wierich and Leuschner,
2006b). In a preliminary experiment the non-hydrolysed lutein ester preparation from Tagetes erecta
(> 60%) was examined in two bacterial toxicity tests (plate incorporation test and preincubation test)
without metabolic activation in test strain TA 100. Ten concentrations each, ranging from 0.316 to
5000 or 0.0316 to 500 μg non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%)/plate
were tested in the plate incorporation test and in the pre-incubation test, respectively. Bacterial toxicity
(scarce background lawn and reduction of the number of revertants) was noted in both tests from
concentrations of 100 μg/plate onwards. Hence, 100 μg/plate was chosen as the top concentration for
the main study. The first experiment of the main study was carried out as a plate incorporation test and
the second as a preincubation test. The supplied test material had a purity of 82.1% and the
concentrations given refer to the active ingredient. Five concentrations ranging from 1 to 100 μg nonhydrolysed lutein ester preparation from Tagetes erecta (> 60%) were employedin the plate
incorporation test and in the pre-incubation test with and without metabolic activation. Bacterial
toxicity (scarce background lawn and reduction of the number of revertants) was observed at the top
concentration of 100 μg non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%) /plate in
all the test strains. No mutagenic effect (no increase in revertant colony numbers as compared with
control) was observed for the non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%)
tested up to a cytotoxic concentration of 100 μg/plate in any of the five test strains in two independent
experiments (plate incorporation and preincubation test) without and with metabolic activation.
A non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%) was assayed for the induction
of gene mutations in cultured mammalian cells (L5178Y TK+/- mouse lymphoma cells) both in the
presence and absence of metabolic activation by a liver post-mitochondrial fraction (S9 mix) from
Aroclor 1254- induced rats (Wierich and Leuschner, 2006c). The study was performed according to
OECD guideline 476 and in compliance with GLP. The test included exposure for 3 and 24 hours
without S9 mix, and a 3-hour exposure with S9 mix, which was carried out twice. The supplied test
item had a purity of 82.1% and the tested concentrations refer to the active ingredient. Tetrahydrofuran
(THF) served as solvent control. No signs of cytotoxicity (decreased survival) were noted in the
preliminary experiment at the top concentration of 2500 μg/ml in the experiments with and without
metabolic activation. As the limit of solubility of the non-hydrolysed lutein ester preparation from
Tagetes erecta (> 60%) in THF was 610 mg/ml the maximum concentration achievable in medium
was 2500 μg/ml. The tests were conducted over a concentration range of 156.3 to 2500 μg/ml. In the
main study, cytotoxicity (decreased survival) was noted immediately after treatment (plating
efficiency step 1) at the top concentration of 2500 μg/ml with or without metabolic activation in the 1st
and 2nd experiment, respectively. No signs of cytotoxicity (decreased survival) were noted in the
following plating for 5-trifluorothymidine (TFT) resistance (plating efficiency step 2) in both
experiments without and with metabolic activation. Methylmethanesulfonate was employed as
positive control in the absence of exogenous metabolic activation and 3-Methylcholanthrene in the
presence of exogenous metabolic activation. The mutation frequency of the solvent controls ranged
from 27.98 to 29.84 per 106 clonable cells in the experiments with and without metabolic activation
and, hence, was well within the historical data-range. The mutation frequencies of the cultures treated
with the non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%) ranged from 17.33 to
37.97 per 106 clonable cells in the experiments with and without metabolic activation. These results
were close to the range of the solvent controls and, hence, no mutagenicity was observed according to
EFSA Journal 2011;9(5):2144
16
Re-evaluation of lutein (E 161b) as a food additive
the criteria for assay evaluation. In addition, no change was observed in the ratio of small to large
mutant colonies, ranging from 1.00 to 1.33 for the cells treated with the non-hydrolysed lutein ester
preparation from Tagetes erecta (> 60%) and from 1.00 to 1.40 for the solvent controls. The positive
controls methylmethanesulphonate (MMS) and 3-methylcholanthrene (3-MC) caused pronounced
increases in the mutation frequency ranging from 149.44 to 641.55 per 106 clonable cells in the case of
MMS and ranging from 176.60 to 223.37 per 106 clonable cells in the case of 3-MC. In addition, the
colony size ratio was moderately shifted towards an increase in small colonies, ranging from 2.50 to
3.00 in case of MMS. The mutation frequency of the solvent controls without and with metabolic
activation for the last 18 experiments (most recent background data) ranges from 6.8 to 47.9 per 106
clonable cells. It was concluded that under the present test conditions, the non-hydrolysed lutein ester
preparation from Tagetes erecta (> 60%), tested up to cytotoxic concentrations in the absence and
presence of metabolic activation in two independent experiments, was negative with respect to the
mutant frequency in the LK5178Y TK+/- mammalian cell mutagenicity test. In an in vivo bonemarrow micronucleus test conducted according to OECD guideline 474 and in compliance with GLP,
groups of 5 rats of each sex received doses of 500, 1000 and 2000 mg non-hydrolysed lutein ester
preparation from Tagetes erecta (> 60%)/kg bw by gavage (Wierich and Leuschner, 2006d). Soybean
oil served as the negative reference item and cyclophosphamide as the positive reference item. No
signs of systemic toxicity were noted after administration of a non-hydrolysed lutein ester preparation
from Tagetes erecta (> 60%) up to the highest dose level of 2000 mg/kg bw. Immediately after
sacrifice, bone marrow smears were prepared. Two sampling times were employed in this study: 24
hours after administration, samples were prepared from the negative reference item, positive reference
item and all doses of test item-treated animals; 48 hours after administration, samples were prepared
only from negative reference item and high dose-treated animals. Two thousand erythrocytes were
evaluated per animal. The highest dose of 2000 mg non-hydrolysed lutein ester preparation from
Tagetes erecta (> 60%) /kg bw, did not result in an increase in the incidence of micronucleated
polychromatic erythrocytes (PCE). The incidences of PCEs for the combined male and female animals
were 1.4 and 1.1 micronuclei per 1000 PCEs for the samples collected 24 and 48 hours, respectively,
after administration of the highest dose (negative reference item: 1.2 and 0.8 micronuclei per 1000
PCE, respectively). Cyclophosphamide resulted in a significant increase to 9.4 micronuclei per 1000
PCEs. The ratio of polychromatic to normochromatic erythrocytes (NCE) was also not influenced by
treatment. It was concluded that under the present test conditions, a non-hydrolysed lutein ester
preparation from Tagetes erecta (> 60%) tested up to the highest dose of 2000 mg/kg bw showed no
mutagenic properties in the rat bone marrow micronucleus test at the two tested sampling times of 24
hours and 48 hours.
The Panel concluded that lutein esters are not of concern with respect to genotoxicity.
3.2.4.
Chronic toxicity and carcinogenicity
In previous evaluations no chronic toxicity/carcinogenicity studies have been described for lutein.
JECFA (2006) described several studies that investigated potential chemopreventive effects of lutein
in mice but these studies have not been included in this opinion because they are not considered
relevant for the evaluation of the safety of lutein.
No long-term or carcinogenicity studies on lutein or lutein esters were submitted since the previous
evaluations.
3.2.5.
Reproductive and developmental toxicity
No multigeneration studies are available. JECFA (2006) described one developmental toxicity study
that has been evaluated by the Panel in the opinion on lutein (EFSA, 2010). The NOAEL in this study
of embryotoxicity/teratogenicity in rats was 1000 mg/kg bw/day, the highest dose tested (Edwards et
al., 2002).
Following a public call for data industry submitted a prenatal developmental toxicity study in which a
non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%) was administered by gavage to
female rats at dose levels of 100, 300 and 1000 mg/kg bw/day from the 6th to 19th day of pregnancy
(Wierich and Leuschner, 2007). The study was performed according to OECD 414 and in compliance
EFSA Journal 2011;9(5):2144
17
Re-evaluation of lutein (E 161b) as a food additive
with GLP. Under the present test conditions, no maternal toxicity was observed at 100, 300 or 1000
mg non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%)/kg bw/day. It was therefore
concluded that the NOAEL for the dams was 1000 mg/kg bw/day, the highest dose tested. There was
no test item-related increase in the incidence of fetal malformations, external, skeletal or soft tissue
variations or skeletal retardations. It was concluded that the non-hydrolysed lutein ester preparation
from Tagetes erecta (> 60%) possessed no teratogenic or embryotoxic properties in rats. The NOAEL
for maternal and developmental toxicity was 1000 mg/kg bw/day, the highest dose tested.
3.2.6.
Human data
The Panel was informed that a number of human trials have been undertaken with a non-hydrolysed
lutein ester preparation from Tagetes erecta (≥ 60%) (Berendschot et al., 2000; Bone et al., 2003;
Bowen et al., 2002; Chung et al., 2004; Heinrich et al., 2003; Koh et al., 2004; Trieschmann et al.,
2007). No adverse effects were reported but the Panel noted that these studies were not designed to
assess the safety of lutein esters.
4.
Discussion
Lutein is a natural carotenoid colour authorised as a food additive in the EU (E 161b) and was
previously evaluated by the SCF in 1975 and JECFA in 2006 and re-evaluated by the ANS Panel in
2010 (EFSA, 2010).
JECFA established a group ADI of 0-2 mg/kg bw/day for lutein from Tagetes erecta and for
zeaxanthin (JECFA, 2006). The SCF could not establish an ADI but concluded that xanthophylls
prepared from natural foods by physical processes are acceptable for use in food (SCF, 1975).
The ANS Panel established an ADI of 1 mg/kg bw/day, based on the NOAEL of 200 mg/kg bw/day
(the highest dose level tested) in a 90-day rat study, and an uncertainty factor of 200 because of the
absence of a multigeneration reproductive toxicity study and of chronic toxicity/carcinogenicity
studies (EFSA, 2010).
The ANS Panel previously noted that this ADI refers to lutein derived from Tagetes erecta containing
≥ 80% carotenoids consisting of lutein and zeaxanthin (79 and 5% respectively) (EFSA, 2010).
According to specifications provided by NATCOL this may refer to the lutein with high
concentrations of total saponified carotenoids at levels of at least 80% (cf. JECFA specifications for
lutein from Tagetes erecta). In the previous opinion re-evaluating the safety of lutein the ANS Panel
concluded that the ADI of 1 mg/kg bw/day does not refer to lutein preparations of lower purity or from
other sources (EFSA, 2010).
The Panel previously also noted that other preparations of lutein are on the market, i.e. lutein with low
concentrations of total carotenoids at levels of ~5-12%, and lutein with high concentrations of total
carotenoids extracted and present as esters at levels of at least 60%. In the previous evaluation (EFSA,
2010) the ANS Panel concluded that the toxicological data base available on these preparations was
too limited to conclude that the ADI also applied to these preparations.
The Panel noted that separate specifications are available for mixed carotenoids, Tagetes extract and
lutein from Tagetes erecta in the JECFA but not in the EU specifications and that the JECFA
specifications on lutein from Tagetes erecta are higher with respect to lutein content (> 70%) than the
EU specifications (> 4%).
Furthermore, the EU specifications and the JECFA specifications (for mixed carotenoids, Tagetes
extract and for lutein from Tagetes erecta) on the purity, differ with respect to solvent residues,
metals, moisture, ash, zeaxanthiwaxes, and the presence or absence of lutein esters Finally, the ANS
Panel previously also noted that the current JECFA ADI for lutein is based on a study using a lutein
product derived from Tagetes erecta with a lutein content of 79%. Also the ADI established recently
by EFSA refers to lutein with high concentrations of total saponified carotenoids at levels of at least
80% (EFSA, 2010). Since in the EU specifications a lutein content of only 4% is required in food
EFSA Journal 2011;9(5):2144
18
Re-evaluation of lutein (E 161b) as a food additive
colour preparations, the material tested may differ substantially from the food colour preparations that
are on the market.
In the present opinion the Panel considered whether the additional studies that were made available by
NATCOL might partially address the gaps identified by the Panel in the toxicological database for
lutein preparations other than lutein with high concentrations of total saponified carotenoids at levels
of at least 80%.
The Panel noted that a specific lutein ester preparation was tested in all additional studies made
available. This was extracted from Tagetes erecta and contained > 60% carotenoid esters (> 93%
lutein esters, remainder zeaxanthin esters). The remaining constituents were long-chain alkanes and
fatty acids plus small amounts of terpenoids and triglycerides. The Panel noted that terpenoids can be
of relevance and therefore the specifications may need to be updated. This preparation will be referred
to subsequently as “a non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%)”.
The Panel has been informed by NATCOL about the nature and levels of the terpenoids that may be
present. Tagetes erecta flowers can contain a variety of terpenoids (Hagers Handbuch der
pharmazeutischen Praxis, 1979). About 38% of the Tagetes erecta flower terpenes is D-limonene and
29.8% are cis- and trans-ocimene, minor terpenes are linalool (5.9%), alpha-D-phellandrene (5.2 %)
and others. These figures are confirmed by other researchers, who also identified piperitone, a
constituent of mint leaves, and caryophyllene, a constituent of cloves and basil, as important
constituents (El-Tantawy et al., 1994; Marotti et al., 2004). Minor amounts of these terpenes may be
present in the > 60% carotenoid esters extract. Being highly volatile substances they are most likely
removed during the manufacturing process together with the extraction solvents hexane and isopropanol. Since the limit value for extraction solvents in E161b is 50 mg/kg NATCOL assumed that
residual terpenes in the carotenoid esters extract do not exceed a low mg/kg level.
The Panel noted that terpenoids may be of toxicological relevance and that therefore specifications
may need to be updated to indicate the type and level of terpenoids present in the preparation.
The Panel has been informed by NATCOL that only hexane and iso-propanol (propan-2-ol) are used
as extraction solvents. The Panel noted that these solvents are already included in the specifications.
No additional data on lutein with low concentrations of total carotenoids at levels of ~5-12% were
provided.
The additional studies included human studies on bioavailability, demonstrating that lutein from lutein
esters is bioavailable (Berendschot et al., 2000; Bowen et al., 2002; Bone et al., 2003; Chung et al.,
2004; Koh et al., 2004; Trieschmann et al., 2007) and that esterification does not impair lutein
bioavailability in humans (Bowen et al., 2002). The Panel concluded that lutein from lutein esters is
bioavailable to an extent that is similar to the bioavailability of lutein itself.
The Panel noted that following passage through the gastrointestinal tract and/or uptake lutein esters are
hydrolysed to form free lutein again.
In the previous opinion on lutein the ANS Panel concluded that based on the studies available there
was no concern with respect to genotoxicity for lutein (EFSA, 2010). Results from three genotoxicity
studies using a non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%) were made
available by industry, including a Salmonella typhimurium reverse mutation assay (Wierich and
Leuschner, 2006b), a gene mutation assay in cultured mammalian cells (L5178Y TK+/- mouse
lymphoma cells) (Wierich and Leuschner, 2006c) and an in vivo bone marrow micronucleus test in the
rat (Wierich and Leuschner, 2006d). No mutagenic activity was seen in any of the tests and the Panel
concluded that lutein esters are not of concern with respect to genotoxicity.
The additional data on the non-hydrolysed lutein ester preparation from Tagetes erecta (> 60%) also
included a 90-day toxicity study performed according to OECD guideline 408 (Wierich and
Leuschner, 2006a), and a reproductive and developmental toxicity study in rats performed according
to OECD 414 (Wierich and Leuschner, 2007); both studies were conducted in compliance with GLP.
No adverse effects were seen in the studies at any dose. The Panel identified for both studies a
NOAEL of 1000 mg/kg bw/day, the highest dose level tested. Given that 6 mg lutein esters (nonEFSA Journal 2011;9(5):2144
19
Re-evaluation of lutein (E 161b) as a food additive
hydrolysed lutein ester preparation from Tagetes erecta (> 60%)) equals 5.5 mg lutein equivalents
(Chung et al, 2004), this NOAEL would amount to 538 mg lutein equivalents/kg bw/day.
The Panel noted that this NOAEL of 538 mg lutein equivalents/kg bw/day is higher than the NOAEL
of 200 mg/kg bw/day (the highest dose level tested) in the 90-day rat study with lutein (260 mg/kg
bw/day of a product consisting of 84% of the carotenoids lutein and zeaxanthin (79 and 5%
respectively) (Kruger et al., 2002) from which the ADI has been derived. The absence of
developmental toxicity at the highest dose of 1000 mg/kg bw/day was also reported for lutein from
marigold extract (79% lutein, 5% zeaxanthin) (Edwards et al., 2002).
Based on these results the Panel concluded that the additional database supports the extension of the
ADI of 1 mg/kg bw/day to lutein with high concentrations of total carotenoids extracted from Tagetes
erecta and present as esters at levels of at least 60%.
The Panel concluded that the ADI of 1 mg/kg bw/day refers to lutein derived from Tagetes erecta
containing at least 80% carotenoids consisting of lutein and zeaxanthin (79 and 5% respectively) and
to lutein with high concentrations of total carotenoids extracted from Tagetes erecta and present as
esters at levels of at least 60%.
The Panel concluded that the toxicological data-base available is too limited to conclude that the ADI
also applies to lutein preparations of lower purity or from other sources.
CONCLUSIONS
The Panel concluded that the additional database supports that the ADI of 1 mg/kg bw/day also refers
to lutein with high concentrations of total carotenoids extracted from Tagetes erecta and present as
esters at levels of ≥ 60%.
The Panel concluded that this ADI refers to lutein derived from Tagetes erecta containing ≥ 80%
carotenoids consisting of lutein and zeaxanthin (79 and 5% respectively) and to lutein with high
concentrations of total carotenoids extracted from Tagetes erecta and present as esters at levels of ≥
60%.
The Panel concluded that the toxicological data-base available is too limited to conclude that the ADI
also applies to lutein preparations of lower purity or from other sources.
EFSA Journal 2011;9(5):2144
20
Re-evaluation of lutein (E 161b) as a food additive
DOCUMENTATION PROVIDED TO EFSA
1. CLG, 1998a. 14 Day Acute Oral Toxicity test in rats on lutein diester Oleoresin. Celsis laboratory
group (CLG) GLP31615. St Louis Missouri. Or: Subacute toxicity with Xangold lutein ester
according to CFR21, Cognis Deutschland GmbH & Co KG, internal report C0500791-0 (1998).
2. CLG, 1998b. Primary eye irritation of lutein diester oleoresin in rabbits. Celsis laboratory group
(CLG) GLP31615. St Louis Missouri. Or: Eye irritation with Xangold lutein ester according to
CFR21, Cognis Deutschland GmbH & Co KG, internal report C0500790-0 (1998).
3. CLG, 1998c. Primary dermal irritation of lutein diester oleoresin in rabbits. Celsis laboratory group
(CLG) GLP31615. St Louis Missouri. Or: Dermal irritation study with Xangold lutein ester
according to CFR21, Cognis Deutschland GmbH & Co KG, internal report C0500789-0 (1998).
4. Wierich P and Leuschner J, 2006a. Repeated dose 90-day oral toxicity study of C-SAT050081 in
rats. LPT Report No. 19586/05. Or: 90 day repeated dose toxicity in rats with Xangold lutein ester
according to OECD 408, Cognis Deutschland GmbH & Co KG, internal report C0501809-0
(2006).
5. Wierich P and Leuschner J, 2006b. Mutagenicity study of C-SAT050081 in the Salmonella
typhimurium reverse mutation assay (in vitro). LPT Report No. 19583/05. Or: Ames test with
Xangold lutein ester according to OECD 471. Cognis Deutschland GmbH & Co KG, internal report
C0501806-0 (2006).
6. Wierich P and Leuschner J, 2006c. Mutagenicity study of C-SAT 050081 in the mouse lymphoma
forward mutation assay –in vitro-. LPT Report No. 19585/05 Or: In vitro mammalian genotoxicity
(mouse lymphoma assay) with Xangold lutein ester according to OECD 476. Cognis Deutschland
GmbH & Co KG, internal report C0501808-0 (2006).
7. Wierich P and Leuschner J, 2006d. Micronucleus test of C-SAT 050081 in bone marrow cells of
the rat by oral administration. LPT Report No. 19584/05. Or: Mammalian chromosome aberration
test (micronucleus test) with Xangold lutein ester according to OECD 476, Cognis Deutschland
GmbH & Co KG, internal report C0501807-0 (2006).
8. Wierich P and Leuschner J, 2007. Prenatal developmental toxicity study of C-SAT 060007 in rats
by oral administration – according to OECD guideline 414 and EC guideline B.31-. LPT Report
No. 19587/05. Or: Developmental toxicity in rats by oral administration with Xangold lutein ester
according to OECD 414, Cognis Deutschland GmbH & Co KG, internal report C0501810-0
(2007).
9. CIIA Communication, 11 April 2011. Preliminary clarification on occurrence data on lutein levels
provided in December 2009.
REFERENCES
Berendschot TTJM, Goldbohm RA, Klopping WA, van de Kraats J, van Norel J and Van Norren D,
2000. Influence of lutein supplementation on macular pigment, assessed with two objective
techniques. Investigative Ophthalmology and Visual Sciences 41, 3322-3326.
Bone RA, Landrum JT, Guerra LH and Ruiz CA, 2003. Lutein and zeaxanthin dietary supplements
raise macular pigment density and serum concentrations of these carotenoids in humans. Journal of
Nutrition 133, 992–998 and Erratum 133, 1953.
Bowen PE, Herbst-Espinosa SM, Hussain EA and Stacewicz-Sapuntzakis M, 2002. Esterification
does not impair lutein bioavailability in humans. Journal of Nutrition 132, 3668–3673.
Chung HY, Rasmussen HM, and Johnson EJ, 2004. Lutein bioavailability is higher from luteinenriched eggs than from supplements and spinach in Men. Journal of Nutrition 134, 1887–1893.
EFSA Journal 2011;9(5):2144
21
Re-evaluation of lutein (E 161b) as a food additive
EC, 2001. Commission of the European Communities (COM). 542 final. Report from the
Commission
on dietary food additive intake in the European Union. Brussels, 01.10.2001.Edwards J, Pfannkuch F
and Marsden E, 2002. Lutein 10% WS (Ro 15-3971/000 — developmental toxicity study by the
oral route (dietary admixture) in the rat (study No. 161/567). Unpublished regulatory document No.
RDR 1008196, dated August 28. Submitted to WHO by Hoffmann-La Roche Ltd., Basle,
Switzerland.
EFSA (European Food Safety Authority), 2006. Opinion of the Scientific Panel on Food Additives,
Flavourings, Processing Aids and Materials in Contact with Food on a request from the
Commission related to Lutein for use in foods for particular nutritional uses. Question N EFSA Q2003-128. Adopted on 26 January 2006.
EFSA (European Food Safety Authority), 2008b. Safety, bioavailability and suitability of lutein for
the particular nutritional use by infants and young children. The EFSA Journal 823, 1-24.
EFSA (European Food Safety Authority), 2009. Safety of use of colouring agents in animal nutrition Part III: ß-apo-8’-carotenal, ethyl ester of ß-apo-8’-carotenoic acid, lutein, zeaxanthin and
concluding remarks. The EFSA Journal 1098, 1-48.
EFSA (European Food Safety Authority), 2010. Scientific Opinion on the re-evaluation of lutein (E
161b) as a food additive. EFSA Panel on Food Additives and Nutrient Sources added to Food
(ANS). EFSA Journal 2010; 8(7): 1678.
El-Tantawy ME, Hamauda MSM and Azzam AS, 1994. Chemical composition and biological activity
of the essential oil of Tagetes erecta L. cultivated in Egypt. Bulletin of Faculty of Pharmacy Cairo
University 32(1), 113-118.
JECFA (Joint FAO/WHO Expert Committee on Food Additives), 2004. WHO/FAO Joint Expert
Committee on Food Additives. Sixty-third meeting Geneva, 8-17 June 2004. Available at:
http://www.who.int/ipcs/publications/jecfa/en/Summary63final.pdf
JECFA (Joint FAO/WHO Expert Committee on Food Additives), 2006. WHO/FAO Joint Expert
Committee on Food Additives. Safety evaluation of certain food additives. WHO Food Additives
Series 54, 49-86. Available at: http://www.inchem.org/documents/jecfa/jecmono/v54je01.pdf
JECFA (Joint FAO/WHO Expert Committee on Food Additives), 2006b. Combined compendium of
food additive specifications - all specifications monographs from the 1st to the 65th meeting (19562005). FAO JECFA Monographs Series, No. 1 Volume 1-3, 2006.
Hagers Handbuch der pharmazeutischen Praxis, 1979. Chemikalien und Drogen, Teil B, R, S,
herausgeg. List von P. H. und Hörhammer L in Gemeinschaft mit H. J. Roth und W. Schmid, Vol
6, p. 5.
Heinrich U, Gärtner C, Wiebusch M, Eichler O, Sies H, Tronnier H and Stahl W, 2003.
Supplementation with b-carotene or a similar amount of mixed caretonoids protects humans from
UV-induced erythema. Journal of Nutrition 133, 98-101.
Koh HH, Murray IJ, Nolan D, Carden D, Feather J and Beatty S, 2004. Plasma and macular responses
to lutein supplement in subjects with and without age-related maculopathy: a pilot study.
Experimental Eye Research 79, 21-27.
Kruger C, Murphy M, DeFreitas Z, Pfannkuch F and Heimbach J, 2002. An innovative approach to
the determination of safety for a dietary ingredient derived from a new source: case study using a
lutein product. Food and Chemical Toxicology 40, 1535-1549.
Marotti M, Piccaglia R, Biavati BB and Marotti I, 2004. Characterization and yield evaluation of
essential oils from different Tagetes species. Journal of Essential Oil Research 16(5), 440-444.
Pfannkuch F, Wolz E, Aebischer CP, Schierle J and Green C, 2000. Ro 15-3971/000 (10%): 13-week
oral toxicity (dietary administration) toxicity study in the rat with a 4-week treatment-free period
EFSA Journal 2011;9(5):2144
22
Re-evaluation of lutein (E 161b) as a food additive
(Roche project 952V99). Unpublished report project No. 161/354 from Covance Laboratories Ltd,
Harrogate UK. Submitted to WHO by Roche, Basle, Switzerland.
Pfannkuch F, Wolz E and Green C, 2001. Ro 15-3971 (10% lutein): Pathological evaluation of the
liver and kidney following a 13-week dietary toxicity study in the rat (report No. 1005032).
Unpublished report No. 0161/424-D6154 from Covance Laboratories Ltd, Harrogate U.K.
Submitted to WHO by Roche, Basle, Switzerland.
SCF, 1975. Reports from the Scientific Committee for Food (1st series). Opinion expressed in 1974.
Food Science and Techniques. Available at:
http://ec.europa.eu/food/fs/sc/scf/reports/scf_reports_01.pdf
TemaNord, 2002. Food additives in Europe 2000; Status of safety assessments of food additives
presently permitted in the EU. TemaNord 2002 560, 188-19.
Tennant, 2006. Screening of colour intakes from non-alcoholic beverages. Report prepared for the
Union of European Beverages Associations (UNESDA).
Trieschmann M, Beatty S, Nolan JM, Hense HW, Heimes B, Austermann U, Fobker M and
Pauleikhoff D, 2007. Changes in macular pigment optical density and serum concentrations of its
constituent carotenoids following supplemental lutein and zeaxanthin: The Luna study.
Experimental Eye Research 84, 718-728.
EFSA Journal 2011;9(5):2144
23
Re-evaluation of lutein (E 161b) as a food additive
GLOSSARY/ABBREVIATIONS
ADI
Acceptable Daily Intake
AFC
Scientific Panel on Additives, Flavourings, Processing Aids and Materials in
Contact with Food
Aluminium
lakes
Aluminium lakes are produced by the absorption of water soluble dyes onto a
hydrated aluminium substrate rendering the colour insoluble in water. The end
product is coloured either by dispersion of the lake into the product or by coating
onto the surface of the product
AMD
Age-related macular degeneration
ANOVA
Analysis of Variance
ANS
Scientific Panel on Food Additives and Nutrient Sources added to Food
ARM
Age-Related Maculopathy
AUC
Area under the curve
CAS
Chemical Abstracts Service
EC
European Commission
EFSA
European Food Safety Authority
EINECS
European Chemical Substances Information System
EXPOCHI
Refers to EFSA Article 36 2008 call for Proposals Focused on Children and Food
Consumption
FAO/WHO
Food and Agriculture Organization/World Health Organization
FEEDAP
Scientific Panel on Additives and Products or Substances use in Animal Feed
FSMPs
Foods for Special Medical Purposes
GLP
Good Laboratory Practise
JECFA
Joint FAO/WHO Expert Committee on Food Additives
3-MC
3-methylcholanthrene
MMS
Methylmethanesulphonate
MPL
Maximum Permitted Limit
MPOD
Macular Pigment Optical Density
NATCOL
Natural Food Colours Association
NCE
Normochromatic erythrocytes
NDA
Scientific Panel on Dietetic products, Nutrition and Allergies
NOAEL
No-Observed-Adverse-Effect Level
OECD
Organisation for Economic Co-operation and Development
PARNUTs
Foods for Particular Nutritional Uses
PCE
Polychromatic erythrocytes
SCF
Scientific Committee on Food
SD
Standard Deviation
EFSA Journal 2011;9(5):2144
24
Re-evaluation of lutein (E 161b) as a food additive
TFT
5-trifluorothymidine
THF
Tetrahydrofuran
TRL
Triglyceride-rich lipoprotein
UNESDA
Union of European Beverage Association
EFSA Journal 2011;9(5):2144
25