1535 PDF

EFSA Journal 2010;8(4):1535
SCIENTIFIC OPINION
Scientific Opinion on the re-evaluation of Brown FK (E 154) as a food

additive1
EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS)2, 3
European Food Safety Authority (EFSA), Parma, Italy
ABSTRACT
The Panel on Food Additives and Nutrient Sources added to Food provides a scientific opinion re-evaluating the
safety of Brown FK (E 154). Brown FK is a food colouring substance permitted only in kippers at a maximum
concentration of 20 mg/kg, which has been previously evaluated by the Joint FAO/WHO Expert Committee on
Food Additives (JECFA) and the EU Scientific Committee for Food (SCF) in 1983. The SCF established an
Acceptable Daily Intake (ADI) of 0.15 mg/kg body weight (bw)/day. JECFA withdrew their temporary ADI of
0.075 mg/kg bw/day for Brown FK at their latest evaluation in 1987, as the toxicological data were considered
inadequate. The Panel was not provided with a newly submitted dossier and based its evaluation on previous
evaluations, additional literature that became available since then and the data available following a public call
for data. The Panel calculated an anticipated refined dietary exposure to Brown FK for European children
ranging from 0.03 to 0.07 mg/kg bw/day in average, and from 0.03 to 0.14 mg/kg bw/day at the 95th percentile.
For the adult population the anticipated mean dietary exposure ranged from 0.002 to 0.07 mg/kg bw/day, and
from 0.007 to 0.14 mg/kg bw/day at the 95th percentile. The Panel noted uncertainties surrounding the NOAELs
reported for Brown FK in a chronic toxicity study and a carcinogenicity study in rats. It also noted the treatmentrelated changes occurring at the highest dose level in the latter study and that, as reported by JECFA,
histopathological examination of the lower groups had not been carried out. The Panel considered, in line with
JECFA, that this examination should have been carried out. The Panel considers therefore that it is unable to
conclude on the safety of Brown FK given the deficiencies in the available toxicity database on the colour.
KEY WORDS
Brown FK, E 154, CAS Registry Number 8062-14-4, food colouring substance, sodium 4-(2,4diaminophenylazo)benzenesulphonate, sodium 4-(4,6-diamino-m-tolylazo)benzenesulphonate, disodium 4,4(4,6-diamino-1,3-phenylenebisazo)dibenzenesulphonate, disodium 4,4-(2,4-diamino-1,3-phenylenebisazo)
dibenzenesulphonate,
disodium
4,4-(2,4-diamino-5-methyl-1,3-phenylenebisazo)dibenzenesulphonate,
trisodium 4,4,4-(2,4-diaminobenzene-1,3,5-trisazo)tribenzenesulphonate.
SUMMARY
Following a request from the European Commission to the European Food Safety Authority, the
Scientific Panel on Food Additives and Nutrient Sources added to Food was asked to deliver a
1
On request, from the European Commission, Question No EFSA-Q-2008-243, adopted on 10 March 2010.
2 Panel members : F. Aguilar, U.R. Charrondiere, B. Dusemund, P. Galtier, J. Gilbert, D.M. Gott, S. Grilli, R. Grtler, J.
Knig, C. Lambr, 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 Acknowledgements: The Panel wishes to thank the members of the Working Group A on Food Additives and Nutrient
Sources of the ANS Panel for the preparation of this opinion: F. Aguilar, N. Bemrah, P. Galtier, J. Gilbert, S. Grilli, R.
Grtler, N-G. Ilbck, C. Lambr, J.C. Larsen, J-C. Leblanc, A. Mortensen, I. Pratt, I. Stankovic, C. Tlustos.
Suggested citation: EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS); Scientific Opinion on the reevaluation of Brown FK (E 154) as a food additive on request from the European Commission. EFSA Journal
2010;8(4):1535. [29 pp.]. doi:10.2903/j.efsa.2010.1535. Available online: www.efsa.europa.eu
European Food Safety Authority, 2010
Re-evaluation of Brown FK (E 154) as a food additive
scientific opinion re-evaluating the safety of Brown FK (E 154) when used as a food colouring
substance.
Brown FK (E 154) is a multi-component mixture of mono-, di- and tri-azo dyes authorised as a food
additive in the EU and previously evaluated by both the EU Scientific Committee for Food (SCF) in
1984 and the Joint FAO/WHO Expert Committee on Food Additives (JECFA) in 1977, 1978, 1985,
1986 and 1987.
Brown FK is described as consisting of a mixture of sodium 4-(2,4-diaminophenylazo)
benzenesulphonate (component I), sodium 4-(4,6-diamino-m-tolylazo)benzenesulphonate (component
II), disodium 4,4-(4,6-diamino-1,3-phenylenebisazo)dibenzenesulphonate (component III), disodium
4,4-(2,4-diamino-1,3-phenylenebisazo)dibenzenesulphonate (component IV), disodium 4,4-(2,4diamino-5-methyl-1,3-phenylenebisazo)dibenzenesulphonate (component V) and trisodium 4,4,4(2,4-diaminobenzene-1,3,5-trisazo)tribenzenesulphonate (component VI), together with subsidiary
colouring matters and with water, sodium chloride and/or sodium sulphate as the principal uncoloured
components.
The Panel noted that according to the CAS Registry Number, Brown FK is comprised of only
components II and IV, rather than the six components mentioned by Commission Directive
2008/128/EC. The Panel also noted that, as reported by Emerton (2008), manufacture of Brown FK
according to the EC specifications is extremely difficult and, given the limited usage of the colour and
the availability of alternative colours, the major manufacturers have stopped offering the colour to EC
specifications. The Panel noted therefore that there is some doubt about whether Brown FK is still
used as a food colour in the European Union.
The SCF in its evaluation of Brown FK in 1984 established an Acceptable Daily Intake (ADI) of 0.15
mg/kg bw/day based on a No-Observed-Adverse-Effect-Level (NOAEL) of 15 mg/kg bw/day
determined in a rat multi-generation study, while JECFA allocated a temporary ADI of 0.075 mg/kg
bw/day for Brown FK based on a 2-year chronic toxicity study in rats. JECFA in its last evaluation in
1987 withdrew its temporary ADI due to the fact that additional data requested by the Committee at
the time of establishing the temporary ADI, namely histopathological examination of the animals in
the low and intermediate dose groups in a rat carcinogenicity study in order to establish a definitive
NOAEL from the study, had not been provided.
The available studies on the absorption, distribution, metabolism and excretion of Brown FK indicate
that the individual components of Brown FK are not absorbed as intact molecules to any extent as
such, but undergo reductive cleavage in the intestine, with some subsequent absorption of aromatic
polyamine metabolic products which are further acetylated before urinary excretion. Metabolites of
Brown FK, including sulphanilic acid, are also excreted in the faeces. Overall given the multicomponent nature of Brown FK, the metabolism of the mixture is complex but the absorption of
metabolites of Brown FK, including aromatic amines, was demonstrated, as evidenced by the toxicity
seen in specific studies with these metabolites.
Adverse effects seen after short-term or prolonged exposure to Brown FK in rats and/or mice include
damage to cardiac (and to a minor extent skeletal) muscle fibres, characterised by vacuolar myopathy,
degenerative lesions including myocardial fibrosis, necrosis and pigment deposition. Other changes
induced by Brown FK include occasional hydropic degeneration of the kidney, fatty change of the
liver, and increases in organ weight (spleen, liver, heart, testes and thyroid). The available studies
provide no clear indication of a NOAEL for the cardiac toxicity or other pathological lesions seen
following oral administration of Brown FK, other than that this NOAEL appears to lie below 100
mg/kg bw/day. In addition to the toxicity findings, pigmentation of tissues was seen from a dose of 55
mg/kg bw/day onwards in mice and 30 mg/kg bw/day onwards in rats. The lowest NOAEL reported
from a dietary study in rats of 15 mg/kg bw/day was based on pigment deposition rather than
biochemical or pathological indicators of toxicity.
In an in vitro bacterial mutagenicity study, different samples of Brown FK have been shown to induce
concentration-dependent frame shift mutations (22- to 50-times the spontaneous mutation frequency)
in Salmonella typhimurium (TA1538). The genotoxicity of samples of Brown FK has also been
demonstrated in fluctuation assays in Salmonella typhimurium using TA1538 and in Escherichia coli
WP2 uvrA, with and without rat liver S9 metabolic activation. The Panel noted that no information
was available on the purity of the material tested in these studies and also noted the absence of in vitro
and in vivo genotoxicity studies on the colour, other than bacterial mutagenicity assays. The Panel also
noted that the different components of Brown FK are metabolised to non-sulphonated aromatic
amines, and that potential impurities in the commercial product include m-phenylenediamine and 4methyl-m-phenylenediamine and other non-specified non-sulphonated aromatic amines, the
genotoxicity of which could be of concern. The Panel additionally noted that 4-methyl-mphenylenediamine (toluene-2,4-diamine) has been assessed by the International Agency for Research
into Cancer (IARC) as a Group 2B carcinogen (possibly carcinogenic to humans). While the Panel
considered that the positive results obtained in in vitro bacterial mutagenicity studies on Brown FK,
together with the potential genotoxicity of its metabolites and possible impurities, give rise to some
concern regarding the genotoxicity of Brown FK, this concern is allayed by the results of long-term
studies in rats and mice showing no convincing evidence of carcinogenicity of Brown FK.
Three long-term toxicological studies have been conducted, one in mice and two in rats. One of the rat
studies was described as a 2-year chronic toxicity study, while the other rat study was a
carcinogenicity study with in utero exposure followed by a 2-year exposure of the offspring. The
mouse study was an 80-week carcinogenicity study. In the mouse study, hepatocellular carcinomas
were observed, however the Panel considered that since the relevance of hepatic tumours in mice for
humans has been questioned, the results of this study are unlikely to be of biological relevance and are
therefore not a suitable basis for risk assessment. No indications for carcinogenicity were obtained in
the long-term study with rats involving in utero exposure. The Panel concluded therefore that these
long-term studies of Brown FK in rats and mice show no evidence of carcinogenic effects relevant to
humans.
No evidence of reproductive toxicity of Brown FK was seen in a multi-generation study and a
developmental toxicity study in rats.
The Panel noted that, in addition to the multi-generation study in rats, used by SCF to derive an ADI,
either the 2-year chronic toxicity study in rats with a reported NOAEL of 15 mg Brown FK/kg bw/day
(used by JECFA to derive their temporary ADI of 0.075 mg/kg bw/day for Brown FK) or the 2-year
carcinogenicity study in rats involving an in utero exposure with a reported (by the authors) NOAEL
of 50 mg/kg bw/day, might be considered to be appropriate studies to use as the basis for establishing
an ADI.
The Panel did not however have access to the reports of any of these studies, and noted the
uncertainties surrounding the NOAELs derived by the authors of the studies, including the relative
contributions of treatment-related changes such as myopathy, myocardial fibrosis and pigment
deposition in establishing effect and no-effect levels. In particular the Panel noted that according to
JECFA, a range of treatment-related effects were reported in the 2-year carcinogenicity study at the
highest dose of 250 mg/kg bw/day. However histopathological examination of the organs and tissues
showing treatment-related changes in the highest dose group had not been carried out in the animals in
the low and intermediate dose groups.
Since treatment-related effects had been observed at the top dose level of 250 mg/kg bw/day in this
study, the Panel considered, in agreement with the view of JECFA, that histopathological examination
of the relevant organs and tissues of (at least) the intermediate dose group animals should have been
carried out. The Panel concluded that given the uncertainties in the available data sets, it was not
possible to derive an appropriate NOAEL from the available data on these studies that could be used
to establish an ADI. Therefore, the Panel is unable to conclude on the safety of Brown FK.
No data on sensitivity to Brown FK are available. Additionally, no cases of intolerance/allergenicity

have been reported after oral exposure to Brown FK, and it appears therefore that at the current levels
of exposure the incidence is very low, if any. On the other hand, the low/absent reports of adverse
clinical reactions after internal Brown FK exposure could be accentuated by the lack of clinical
awareness of this possibility.
Based on the fact that Brown FK is permitted only in kippers at a maximum concentration of 20
mg/kg, the Panel calculated an anticipated refined dietary exposure (Tier 2) for European children
(aged 1-10 years and weighing 15-30 kg) ranging from 0.03 to 0.07 mg/kg bw/day in average, and
from 0.03 to 0.14 mg/kg bw/day at the 95th percentile. For the adult population the anticipated mean
dietary exposure ranged from 0.002 to 0.07 mg/kg bw/day, and from 0.007 to 0.14 mg/kg bw/day at
the 95th percentile.
The Panel notes that the JECFA specification for lead in Brown FK is < 2 mg/kg, whereas the EC
specification is < 10 mg/kg.
The Panel also notes that the aluminium lake of the colour could add to the daily intake of aluminium,
for which a Tolerable Weekly Intake (TWI) of 1 mg aluminium/kg bw/week has been established, and
that therefore specifications for the maximum level of aluminium in the lakes may be required.
TABLE OF CONTENTS
Abstract .................................................................................................................................................... 1
Key words ................................................................................................................................................ 1
Summary .................................................................................................................................................. 1
Table of contents ...................................................................................................................................... 5
Background as provided by the European Commission........................................................................... 6
Terms of reference as provided by the European Commission ................................................................ 6
Assessment ............................................................................................................................................... 7
1. Introduction ..................................................................................................................................... 7
2. Technical data .................................................................................................................................. 7
2.1.
Identity of the substance ......................................................................................................... 7
2.2.
Specifications .......................................................................................................................... 8
2.3.
Manufacturing process .......................................................................................................... 10
2.4.
Methods of analysis in foods ................................................................................................ 10
2.5.
Reaction and fate in food ...................................................................................................... 10
2.6.
Case of need and proposed uses............................................................................................ 11
2.7.
Information on existing authorisations and evaluations........................................................ 11
2.8.
Exposure ............................................................................................................................... 11
3. Biological and toxicological data .................................................................................................. 12
3.1.
Absorption, distribution, metabolism and excretion ............................................................. 12
3.2.
Toxicological data ................................................................................................................. 14
3.2.1. Acute oral toxicity ............................................................................................................ 14
3.2.2. Short-term and subchronic toxicity .................................................................................. 14
3.2.3. Genotoxicity ..................................................................................................................... 17
3.2.4. Chronic toxicity and carcinogenicity ................................................................................ 18
3.2.5. Reproductive and developmental toxicity ........................................................................ 19
3.2.6. Allergenicity, hypersensitivity and intolerance ................................................................ 20
4. Discussion ...................................................................................................................................... 20
Conclusions ............................................................................................................................................ 23
Documentation provided to EFSA ......................................................................................................... 24
References .............................................................................................................................................. 24
Glossary and abbreviations .................................................................................................................... 29
BACKGROUND AS PROVIDED BY THE EUROPEAN COMMISSION

According to the Framework Directive 89/107/EEC4 on food additives, the Scientific Committee on
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 authorisation, have been evaluated for their safety by the SCF or by its successor the
European Food Safety Authority (EFSA).
Directive 89/107EEC 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.
In accordance with Regulation (EC) No 1333/2008, the Commission should, after consultation with
EFSA, set up by 20 January 2010 an evaluation programme for EFSA to re-evaluate the safety of the
permitted food additives. That programme will define the needs and the order of priorities according to
which the approved food additives are to be examined.
Food colours were among the first additives to be evaluated, therefore many of the evaluations are old.
For some of these colours many new studies have become available and the results of these studies
should be included in the evaluation. Therefore, food colours should be evaluated with priority. The
order of priorities for the re-evaluation of the remaining permitted food additives will be set in the
Regulation for the re-evaluation program.
TERMS OF REFERENCE AS PROVIDED BY THE EUROPEAN COMMISSION

The Commission asks the European Food Safety Authority to start a systematic re-evaluation of
authorised food additives and to issue scientific opinions on these additives, taking into account that
colours as a group should be given the highest priority for the reasons outlined above.
4
5
OJ L 40, 11.2.1989, p. 27
OJ L 354, 31.12.2008, p. 16.
ASSESSMENT
1.
Introduction
The present opinion deals with the re-evaluation of the safety of Brown FK (E 154) when used as a
food colouring substance.
Brown FK (E 154) is an azo-dye authorised as a food additive in the EU and previously evaluated by
the Joint FAO/ WHO Expert Committee on Food Additives (JECFA) (JECFA, 1977; 1978; 1985;
1986, 1987). JECFA withdrew their previously established temporary Acceptable Daily Intake (ADI)
of 0.075 mg/kg bw/day for Brown FK at their latest evaluation (JECFA, 1987) due to the fact that
additional data requested by the Committee at its previous evaluation had not been provided. The EU
Scientific Committee for Food (SCF) evaluated Brown FK in 1983 and established an ADI of 0.15
mg/kg bw/day (SCF, 1984). Brown FK has also been reviewed by TemaNord (TemaNord, 2002).
The Panel was not provided with a newly submitted dossier and based its evaluation on previous
evaluations, additional literature that became available since then and the data available following a
public call for data. The Panel noted that not all original studies on which previous evaluations were
based were available for re-evaluation by the Panel.
2.
Technical data
2.1.
Identity of the substance
Brown FK (E154) is a red-brown azo-dye which is reported to be a mixture of six mono-, di- and triazo dyes. The six individual components have the following names, according to Commission
Directive 2008/128/EC6 on purity criteria concerning colours for use in foodstuff:
I
II
III
IV
V
VI
sodium 4-(2,4-diaminophenylazo)benzenesulphonate - C12H11N4NaO3S

sodium 4-(4,6-diamino-m-tolylazo)benzenesulphonate - C13H13N4NaO3S
disodium 4,4-(4,6-diamino-1,3-phenylenebisazo)dibenzenesulphonate
- C18H14N6Na2O6S2
disodium 4,4-(2,4-diamino-1,3-phenylenebisazo)dibenzenesulphonate
- C18H14N6Na2O6S2
disodium 4,4-(2,4-diamino-5-methyl-1,3-phenylenebisazo)dibenzenesulphonate
- C19H16N6Na2O6S2
trisodium 4,4,4-(2,4-diaminobenzene-1,3,5-trisazo)tribenzenesulphonate
- C24H17N8Na3O9S3
Brown FK is soluble in water and sparingly soluble in ethanol (JECFA, 2006).

The CAS Registry Number 8062-14-4, which is used in the JECFA specifications, refers to a mixture
of the compounds II and IV, that also have individual CAS Registry Numbers (II: 6300-61-4; IV:
14515-08-3) whereas no CAS Registry Numbers were found for the other components.
At least five synonyms for Brown FK are in use (ChemIDplus advanced, via internet, 2010). The most
commonly used synonym in published literature is CI Food Brown 1.
Commission Directive 2008/128/EC of 22 December 2008 laying down specific purity criteria concerning colours for use in
foodstuffs. Official Journal of the European Communities, L 6, 10.1.2009.
The structural formulae of the six individual components of the Brown FK mixture are as follows:
Na+
O
O
NH2
H3C
Na+
S
O
NH2
VI
Na+
NH2
Na+
NH2
Na+
Figure 1: Structural formulae of the six individual components of the Brown FK mixture
2.2.
Specifications
Specifications have been defined in the Commission Directive 2008/128/EC and by JECFA (JECFA,
2006) (Table 1).
4,4-(2,4-diamino-1,3-phenylenebisazo)dibenzenesulphonate (component IV), disodium 4,4-(2,4EFSA Journal 2010;8(4):1535
diamino-5-methyl-1,3-phenylenebisazo)dibenzenesulphonate (component V) and trisodium 4,4,4(2,4-diaminobenzene-1,3,5-trisazo)tribenzenesulphonate (component VI), together with subsidiary
colouring matters, water, sodium chloride and/or sodium sulphate as the principal uncoloured
components. Brown FK is described as the sodium salt. The calcium and the potassium salts are also
permitted (Directive 2008/128/EC).
The purity is specified as not less than 70% total colouring matters. Of the total colouring matters
present, the specifications stipulate that the proportions of the components of the mixture shall not
exceed 26% for component I, 17% each for components II and III, 16% for component IV, 20% for
component V and 16% for component VI. The remaining 30% may be accounted for by sodium
chloride or sodium sulphate, although this is never mentioned explicitly, together with < 3.5%
subsidiary colouring matters, 0.7% 4-aminobenzene-1-sulfonic acid, 0.35% m-phenylenediamine
and 4-methyl-m-phenylenediamine, 0.0007% unsulphonated primary aromatic amines other than mphenylenediamine and 4-methyl-m-phenylenediamine and 0.2% water insoluble material.
Specifications for metals are also provided.
According to Emerton (2008), manufacture of Brown FK to the EC specifications is extremely
difficult and, given the limited usage of the colour and the availability of alternative colours, the major
manufacturers have stopped offering the colour according to EC specifications.
Table 1:
Specifications for Brown FK according to Commission Directive 2008/128/EC and

JECFA (2006)
Purity
Water insoluble matter
Subsidiary colouring matters
Organic compounds other than colouring
matters:
- 4-aminobenzene-1-sulfonic acid
- m-phenylenediamine
and
4-methyl-mphenylenediamine
Unsulphonated primary aromatic amines other
than m-phenylenediamine and 4-methyl-mphenylenediamine
Ether extractable matter
Arsenic
Lead
Mercury
Cadmium
Heavy metals (as Pb)
Commission Directive
2008/128/EC
0.2%
3.5%
JECFA, 2006
0.7%
0.35%
0.7%
0.35%
0.007% (calculated as
aniline)
0.007% (calculated as
aniline)
From a solution of pH 7
0.2%
3 mg/kg
10 mg/kg
1 mg/kg
1 mg/kg
40 mg/kg
0.2%
02%
3.5%
2 mg/kg
-
JECFA revised their earlier specifications for Brown FK in 1987, to provide those outlined above, at
the same time that they withdrew the temporary ADI, in order to facilitate the identification of
appropriate materials to be used in further toxicological studies (JECFA, 1987).
The Panel noted that the specifications on the purity of Brown FK would permit concentrations of
unsulphonated aromatic amines other than m-phenylenediamine and 4-methyl-m-phenylenediamine of
up to 70 mg/kg in the colour. Given the maximal allowed concentration of Brown FK that can be
added to food (20 mg/kg food), the concentration of these unidentified unsulphonated primary
aromatic amines in food could be 1.4 g/kg food. The Panel additionally noted that the specifications
on the purity of Brown FK allow the aromatic amines m-phenylenediamine and 4-methyl-mphenylenediamine to be present in Brown FK at a maximum concentration of up to 3500 mg/kg, and
that 4-methyl-m-phenylenediamine (toluene-2,4-diamine) has been assessed by the International
Agency for Research into Cancer (IARC) as a Group 2B carcinogen (possibly carcinogenic to
humans). Given the maximal allowed concentration of Brown FK that can be added to food (20 mg/kg
food) the concentration of these compounds in food may reach 70 g/kg food.
The Panel noted that the JECFA specification for lead is 2 mg/kg, whereas the EC specification is
10 mg/kg.
No information is available indicating that Brown FK is used in aluminium lake form, although
according to EU legislation this is allowed. According to Commission Directive 2008/128/EC, the
above purity criteria for the pure substance also apply to the raw material from which the aluminium
lake is produced. In addition, the aluminium lake should contain no more than 0.5% hydrochloric acid
(HCl)-insoluble material, and no more than 0.2% ether-extractable material, under neutral conditions.
There are no additional specification requirements for the aluminium lake.
JECFA does not give specifications for aluminium lakes of Brown FK, other than reference to the
General Specifications for Aluminium Lakes of Colouring Matters (JECFA, 2004). The Brown FK
used in the production process should comply with the specifications as given above and the
aluminium lake should contain not more than 2% water-soluble chlorides and sulphates calculated as
sodium salts, not more than 0.5% HCl-insoluble matter, 0.2% ether-extractable matter, 3 mg
arsenic/kg and 5 mg lead/kg. Unreacted aluminium oxide may also be present in the final product (not
specified).
The Panel notes that the aluminium lake of the colour could add to the daily intake of aluminium, for
which a Tolerable Weekly Intake (TWI) of 1 mg aluminium/kg bw/week has been established (EFSA,
2008), and that therefore specifications for the maximum level of aluminium in the lakes may be
required.
2.3.
Manufacturing process
According to JECFA, Brown FK is prepared by coupling diazotised sulphanilic acid with a mixture of
m-phenylenediamine and 4-methyl-m-phenylenediamine (JECFA, 1985). Brown FK may be
converted to the corresponding aluminium lake under aqueous conditions by reacting aluminium oxide
with the colouring matter. Undried aluminium oxide is usually freshly prepared by reacting aluminium
sulphate or aluminium chloride with sodium carbonate or sodium bicarbonate or aqueous ammonia.
Following lake formation, the product is filtered, washed with water and dried (JECFA, 2004).
2.4.
Methods of analysis in foods
Several methods for the determination of Brown FK in foods are described in the literature, of which
variations of High Pressure Liquid Chromatography (HPLC) appear to be most generally employed.
2.5.
Reaction and fate in food
In general, the majority of colour additives are unstable in combination with oxidising and reducing
agents in food. Since colour depends on the existence of a conjugated unsaturated system within the
dye molecule, any substance which modifies this system (e.g. oxidising or reducing agents, sugars,
acids, and salts) will affect the colour (Scotter and Castle, 2004).
10
2.6.
Case of need and proposed uses
Permitted use levels have been defined in the Directive 94/36/EC7 on colours for use in foodstuffs.
Currently, Brown FK is an authorised synthetic food colouring substance in the EU, exclusively
permitted with a maximal allowed use level of 20 mg/kg food in kippers (Directive 94/36/EC). No
data on actual levels of use of Brown FK are available. As reported by Emerton (2008), manufacture
of Brown FK according to the EC specifications is extremely difficult, and given the limited usage of
the colour and the availability of alternative colours, the major manufacturers have stopped offering
the colour to EC specifications. The Panel noted therefore that there is some doubt about whether
Brown FK is still used as a food colour in the European Union.
2.7.
Information on existing authorisations and evaluations
Brown FK is permitted as a food additive in the EU under Directive 94/36/EC.

Brown FK (E 154) has been evaluated by JECFA in 1977 (JECFA, 1977, 1978), when no ADI was
established, and in 1985, when a temporary ADI of 0.075 mg/kg bw/day was established (JECFA,
1985, 1986). JECFA withdrew the ADI for Brown FK at their latest evaluation (JECFA, 1987) as the
toxicological data were considered inadequate. The SCF evaluated Brown FK in 1983 and established
an ADI of 0.15 mg/kg bw/day (SCF, 1984). Brown FK has also been reviewed by TemaNord, who
recommended that the colour should be re-evaluated if it was still in use, although it noted that
exposure was low (TemaNord, 2002).
2.8.
Exposure
Because Brown FK is only allowed to be present in a single specified foodstuff, kippers, estimation of
exposure based on the Budget method was considered inappropriate by the Panel. As no reported or
actual levels of use or observed analytical data were provided to the Panel, only the Tier 2 approach
has been assessed. Dietary exposure estimates have been performed for children and for the adult
population based on the maximum level of use of 20 mg/kg as specified in the EC legislation on food
colours (Directive 94/36/EC) and the individual food consumption data available for consumers of
kippers.
Exposure estimates of Brown FK for children (aged 1-10 years and weighing 25-30 kg) have been
performed by the Panel based on detailed individual food consumption data from seven European
countries (Belgium, France, the Netherlands, Czech Republic, Italy, Finland, Greece), provided by the
EXPOCHI (Individual food consumption data and exposure assessment studies for children)
consortium (Huybrechts et al., in press). As UK is not part of the EXPOCHI consortium, estimates for
UK children (aged 1.5-4.5 years) were made by the Panel based on the UK detailed individual food
consumption data (UK NDNS, 1992-1993) available from the Union of European Beverage
Association (UNESDA) report (Tennant, 2006). In all these surveys, the percentage of consumers (of
kippers) was lower than 3% and the reported consumption ranged from 32 to 84 g/day for average
consumers, and from 23 to 170 g/day at the 95th percentile of consumers.
Estimates of Brown FK for the adult population (>18 years old) have been made by the Panel with the
available data from the EFSA Concise European Food Consumption Database which gives access to
aggregate data on the fish and fish product categories consumed by 15 European countries. These
categories were used since individual consumption data were not available for the adult population,
but provide a conservative estimate of exposure. The percentage of consumers ranged between 5 to
7
European Parliament and Council Directive 94/36/EC of 30 June 1994 on colours for use in foodstuffs. Official Journal of the European
Communities, L 237, 10.9.94.
11
85% and the reported consumption ranged from 7 to 198 g/day for average consumers and from 20 to
420 g/day at the 95th percentile of consumers.
Table 2 summarises the anticipated dietary exposure to Brown FK in children and in the adult
population.
Anticipated mean dietary exposure, based on the maximum level of use of 20 mg/kg (Tier 2), for
European children (aged 1-10 years and weighing 15-30 kg) was estimated to be between 0.03 and
0.07 mg/kg bw/day and ranged from 0.03 to 0.14 mg/kg bw/day at the 95th percentile.
For the adult population (aged >18 years and weighing 60 kg), anticipated mean dietary exposure was
estimated to be between 0.002 and 0.07 mg/kg bw/day and ranged from 0.007 to 0.14 mg/kg bw/day at
the 95th percentile.
Table 2: Summary of anticipated exposure to Brown FK in children and adult populations

Adult population
(>18 years old)
Tier 2. Maximum Permitted Level
Mean exposure (range)
Exposure 95th or 97.5th percentile
3.
0.002-0.07
0.007-0.14
Child population
(1-10 years old)
mg/kg bw/day
0.03-0.07
0.03-0.14
Biological and toxicological data
No toxicological or biological information was submitted for the re-evaluation of Brown FK following
a public call for data, prior to the start of this re-evaluation.
A literature search was conducted on the most commonly available online databases for toxicological
and biological information (PubMed, Science Direct, Toxline and Web of Knowledge), to cover recent
published literature on Brown FK. However, very little new information was identified.
The present opinion briefly summarises the major toxicological studies on Brown FK evaluated
previously by JECFA (JECFA, 1977, 1985), the SCF (SCF, 1984) and TemaNord (2002). The Panel
did not have access to the original reports referenced in those evaluations. The Panel noted that many
of the studies on Brown FK were carried out before the introduction of Good Laboratory Practice
(GLP) in 1981.
The Panel also noted that Brown FK is a complex mixture of different coloured components, and that
little or no information was available in some of the toxicological and biological studies about the
composition of the material tested.
3.1.
Absorption, distribution, metabolism and excretion
The JECFA (e.g. JECFA, 1985) evaluations describe a number of studies on the toxicokinetics of
Brown FK. The main conclusions of the JECFA evaluations were that the parent compound is
absorbed only to a very limited extent but undergoes reductive cleavage in the intestine, with some
absorption of aromatic amine metabolic products which are further acetylated before urinary
excretion. Metabolites of Brown FK including sulphanilic acid are also excreted in the faeces. Overall
given the multi-component nature of Brown FK, the metabolism of the mixture is complex, but
absorption of some metabolites of Brown FK, including aromatic amines, was demonstrated, as
evidenced by toxicity seen in specific studies with these metabolites.
12
A series of in vitro studies (Fore et al., 1967; Fore and Walker, 1967; Walker, 1968) demonstrated that
Brown FK undergoes azo reduction in the presence of rat ileal and caecal contents. In vitro
metabolism of Brown FK (assumed to be a mixture of its component colours) resulted in formation of
sulphanilic acid and a phenazine-like material, identified as a complex mixture containing mainly
1,4,7-triaminophenazine, a methyl-1,4,7-triaminophenazine and some other ill-defined products. Four
individual components of Brown FK (components I, II and IV and one undefined) were also
demonstrated to undergo azo reduction. When Brown FK was incubated with rat liver homogenate,
azo reduction also occurred but the phenazine-related degradation material could not be detected.
After oral administration of Brown FK to rats, mice, rabbits and guinea-pigs, colouration was seen in
the faeces (up to 24 hours after treatment) and urine (within 15 minutes after treatment). Sulphanilic
acid was identified in urine and faeces (Fore et al., 1967). The phenazine-related material identified in
vitro was also detected in vivo, mainly in caecal contents (predominantly during the first six hours
after dosing), and was also detectable in trace amounts in the faeces. In addition, a "blue material" was
excreted in the urine. Walker (1968) identified the "blue material" as an indamine (not specified),
formed from the amines initially produced by azo reduction of the mono azo components (I and II) of
Brown FK and postulated that indamine was an intermediate in the formation of the phenazine-related
products.
The metabolism of components I and II of Brown FK (sodium 4-(2,4-diaminophenylazo)
benzenesulphonate and sodium 4-(4,6-diamino-m-tolylazo)benzenesulphonate) was further
investigated in the rat (Unilever, 1969a; 1969c; 1969d). The metabolism of these two components was
qualitatively similar, with the majority of the colour being reductively cleaved to sulphanilic acid and
the related aromatic polyamines, which were acetylated before excretion in urine, although traces of
the colour were excreted unchanged in the faeces. The authors postulated that the metabolism of other
Brown FK components was likely to be similar, and that the primary metabolic reactions would be
products of cleavage of the azo linkages (Unilever, 1969a; 1969d). According to JECFA, the six major
metabolites (besides sulphanilic acid) of the individual components of Brown FK would be:
NH2
NH2
NH2
H2N
NH2
NH2
NH2
H2N
H2N
CH3
1,2,4-triaminobenzene
NH2
1,2,4,5-tetraminobenzene
NH2
NH2
NH2
2,4,5-triaminotoluene
H3C
NH2
NH2
1,2,3,4-tetraminobenzene
NH2
NH2
H2N
NH2
NH2
NH2
2,3,4,5-tetraminotoluene
NH2
NH2
1,2,3,4,5-pentaminobenzene
Figure 2: The major metabolites of the six individual components of Brown FK (in addition to
sulphanilic acid)
Both 1,2,4-triaminobenzene (metabolite A, from component I) and 2,4,5-triaminotoluene (metabolite
B from component II) have been shown to uncouple oxidative phosphorylation in vitro, to interfere
with ATP production in the muscle cell, and to lead to ionic imbalance with cell death (Unilever,
1971c).
13
In a study in which component I was administered to human subjects, no unchanged colour was
detected in the urine (Unilever, 1971b). In addition, no substantial urinary excretion of sulphanilic acid
was noted. The authors postulated that orally-administered sulphanilic acid is not absorbed in humans
and is probably excreted in the faeces. However, this was not confirmed experimentally (Unilever,
1971b).
No additional data on absorption, distribution, metabolism and excretion of Brown FK are provided in
the SCF and TemaNord evaluations, and no new studies have been published since these evaluations
and those of JECFA.
3.2.
Toxicological data
3.2.1.
Acute oral toxicity
The JECFA evaluations contain information on the acute toxicity of Brown FK. LD50 values range
from 960-1149 mg/kg bw in mice, 780-970 mg/kg bw in rats, 2610 mg/kg bw in guinea-pigs and 390590 mg/kg bw in rabbits (Unilever, 1966b; Grasso et al., 1968a).
No additional data on acute toxicity were provided in the SCF (1984) and TemaNord (2002)
evaluations.
3.2.2.
Short-term and subchronic toxicity
The JECFA evaluations (1977, 1978, 1985, 1986, 1987) describe a number of subchronic studies on
Brown FK (commercial product, proportions of the individual components not generally specified)
conducted in mice, rats and pigs. In addition, subchronic studies are described in which some of the
individual components of Brown FK or its amine metabolites were investigated.
No additional data are provided in the SCF (1984) and TemaNord (2002) evaluations and no new
published studies have been identified since these previous evaluations.
A number of the studies described by JECFA involved intravenous or intraperitoneal administration of
Brown FK, its individual components or metabolites, and while these provide some useful information
on the toxicity of the colour, they have in the main not been included in the summary below, except
where considered to be of particular relevance.
A characteristic finding in subchronic and chronic toxicity studies with Brown FK, administered by
the oral route, was the deposition of a brown pigment in various tissues (heart, muscle, thyroid,
intestine, stomach and brains). Attempts have been made in various studies to identify this pigment but
results have been inconclusive. Although it has been proposed that it may be 1,4,7-triaminophenazine,
one of the phenazine-related degradation products of Brown FK, which is brown in colour and waterinsoluble, some of the early publications report the pigment to be lipofuscin.
Studies in rats
Groups of rats (Carworth Farm E strain, 5 males and 5 females/group) received Brown FK orally by
gavage at doses of 0, 100, 250, 500, 1000, 1250, 1500, or 2000 mg/kg bw/day for up to 43 days. Oral
treatment with doses of 500 mg/kg bw/day and higher resulted in weight loss and damage to cardiac
and skeletal muscles, characterised by vacuolar myopathy and pigment deposition, described as
14
lipofuscin. No such effects were seen after intraperitoneal administration of Brown FK suggesting that
intestinal breakdown products are responsible for the observed lesions (Grasso et al., 1968a).
The same publication (Grasso et al., 1968a) reports the result of a study in which rats were fed 500
mg/kg bw/day of three individual components of Brown FK (components I and II are mentioned by
JECFA (1985); the identity of the third component is not given). In this study, components I and II
also produced muscle damage, the response produced by component I being very similar to that of
Brown FK mixture. Component II produced vacuolar myopathy in heart and skeletal muscles as well
as lipofuscin formation, but the incidence of lesions was considerably lower than that produced by
component I.
Rats (weanling; 24) were either given Brown FK in the diet at a level of 2% (equivalent to 1000 mg/kg
bw/day) for 12 weeks, or received Brown FK by gavage, also at doses of 1000 mg/kg bw/day (2-3
doses as a 10% solution). In animals given Brown FK in the diet, fibrillolysis and an increase in
number and electron-density of lysosomes were observed in cardiac muscle fibres during weeks 2-3 of
the study. These changes were accompanied by a marked elevation of acid phosphatase. As described
by the authors, progressive deposition of pigment, described as lipofuscin, in cardiac and skeletal
muscles was the principal pathological feature during weeks 3-12. In animals receiving Brown FK by
gavage, myopathy and fibrillolysis of the cardiac and skeletal muscles was also noted, accompanied by
a moderate increase in acid phosphatase activity and a loss of phosphorylase activity (Grasso et al.,
1968b).
Groups of weanling rats (Colworth-Wistar, 12/sex/group) were fed Brown FK (51% coloured
components, 47% salt) for 112 days. Dietary levels of coloured components were approximately
0.025, 0.05, 0.25, 0.5, or 1%, equivalent to approximately 12.5, 25, 125, 250, or 500 mg/kg bw/day.
Controls received non-treated diet while a further control group (n=24) received a diet containing 1%
sodium chloride. Tissue pigmentation (not further specified) was observed at 125 mg/kg bw/day;
while at 250 mg/kg bw/day liver enlargement was noted and at 500 mg/kg bw/day there was reduced
growth, poor food utilisation, enlargement of the liver, testes, and thyroid, histological evidence of
myopathy and myocardial fibrosis, increased urinary indican (indoxyl sulphate and indoxyl
glucuronate) excretion, and elevation of aspartate aminotransferase (AST). The authors considered
that based on these data the no-effect level was 25 mg/kg bw/day (Unilever, 1966a).
Brown FK (purity 80%; no detail on proportion of components) was administered to rats at dietary
levels of 0, 0.001, 0.01, 0.1, or 1% (equivalent to 0, 0.5, 5, 50, or 500 mg/kg bw/day) for 150 days. At
a dose of 500 mg/kg bw/day typical myocardial changes were noted in a single male rat. At this dose
pigment deposition (site of deposition not specified, described as lipofuscin) was also noted
(especially in females). The authors considered that no adverse effects were noted on growth, food
consumption, haematological indices, liver and kidney function, or organ weights. The authors
determined a No-Observed-Adverse-Effect-Level (NOAEL) of 50 mg/kg bw/day (Gaunt et al., 1968).
Groups of rats (10-12/group) were given Brown FK by oral gavage at doses of 0, 100 or 1000 mg/kg
bw/day, or by intraperitoneal injection at doses of 0, 100, 250, or 1000 mg/kg bw/day for up to a
maximum of 43 days. After oral gavage with 100 mg/kg bw/day, two rats showed histological
evidence of pericardial and sub-pericardial lesions. In addition, early hydropic degeneration of the
kidney was seen in two rats, with one of these animals also showing fatty change in the liver. At 1000
mg/kg bw/day, marked cardiac damage was seen. After intraperitoneal administration, Brown FK
produced little or no cardiac damage at any dose tested. At the highest oral dose of 1000 mg/kg
bw/day, most animals showed congestion, fatty change and/or necrosis of the liver with hydropic
degeneration. There was no obvious splenomegaly (BIBRA, 1964).
Rats (20/group) were administered Brown FK orally or by intraperitoneal injection (8/group) at a dose
level of 500 mg/kg bw/day for three weeks. Of the orally-treated animals, six animals died between
days 5 and 11 of the daily dosing regime. Post-mortem examination revealed general tissue staining in
most animals. Myopathy and myocardial fibrosis were seen in 8 hearts out of 18 examined
15
histologically, and a brown pigment was observed in small amounts in nine hearts after three weeks. In
rats given Brown FK intraperitoneally, none of the treated animals died during the treatment period.
At post-mortem examination, general organ staining was observed in all animals. Hearts from seven
rats were examined microscopically and degenerative lesions were found in one heart and small
amounts of pigment in three hearts after three weeks (Unilever, 1968).
Groups of rats (Colworth-Wistar, 3/sex/group) were fed component I or component II at dietary levels
of 0, 0.5, or 1% (equivalent to 0, 250, or 500 mg/kg bw/day) for six weeks. After treatment with
component I, staining of the heart, thyroid, muscle, brain and intestine due to pigment deposition was
noted. No information was provided on the dose-response relationship. After treatment with
component II, pale hearts and meningeal haemorrhage were seen, deposition of pigmentation was
similar to component I. Again, no information was provided on the dose-response relationship.
Histologically, more pigment was seen in rats treated with component I compared to component II.
Myopathy and myocardial fibrosis were observed in one animal treated with component I and four
animals treated with component II (no information on dose administered) (Unilever, 1968).
Groups of rats (6-7/group) received 1,2,4-triaminobenzene (a metabolite A of Brown FK) orally at 0,
50, 60, 75, or 100 mg/kg bw/day for two weeks (5 days/week). In the 100 mg/kg bw/day group, 6 out
of 7 animals died after three doses, and post-mortem examination revealed severe myopathy and
myocardial fibrosis. Of the animals treated with 75 mg/kg bw/day, 7 out of 11 also died after three
doses (no detail on pathology). Heart pigmentation was seen after five or more days of treatment (no
detail on dose). Animals receiving lower doses of 1,2,4-triaminobenzene showed both extensive heart
pigmentation and cardiac necrosis (Unilever, 1969b).
Groups of rats (6/group) were orally administered 1,2,4,5-tetraminobenzene (metabolite C) at 0, 150
or 200 mg/kg bw/day, or 1,2,3,4-tetraminobenzene (metabolite D) at 0, 125 or 166 mg/kg bw/day for
two weeks (5 days/week). No obvious myopathy and myocardial fibrosis were observed, and only
instances of diffuse increase in interstitial cells in the heart were noted. No heart or thyroid pigment
deposition was observed (Unilever, 1969b).
Studies in mice
Groups of 6-week old mice (Colworth C57Bl, 10/sex/group) were fed Brown FK (51% coloured
components, 47% salt, no detail on proportions of different components) for 90 days at Brown FK
levels in the diet of approximately 0, 0.025, 0.0375, 0.05, 0.125, 0.25, 0.375, 0.5, or 1% of the diet
(equivalent to approximately 0, 35, 55, 70, 180, 360, 535, 715, or 1430 mg/kg bw/day) (Unilever,
1958). A further control group (20 animals) received a diet containing 1% sodium chloride. Pigment
deposition in the tissues was noted at dose levels of 180 mg Brown FK/kg bw/day and above, while
splenic enlargement occurred at dose levels of 360 mg/kg bw/day and above, and the liver and heart
were enlarged at 715 mg/kg bw/day and above. At 1430 mg/kg bw/day there was reduced growth,
poor food utilisation, enlargement of liver, spleen, heart and testicles, and histological evidence of
myopathy and myocardial fibrosis.
Groups of mice (Colworth C57Bl; 3/sex/group) were fed the pure Brown FK components I or II at
dietary levels of 0, 0.5, or 1% (equivalent to approximately 0, 715, or 1430 mg/kg bw/day) for six
weeks. Darkening of the thyroid was seen with both components and the intestines and squamous
portions of the stomach were stained salmon-pink (no detail on dose-response relationship). More
pigment was seen in mice fed component I compared to component II. At a dose of 1430 mg/kg
bw/day, myopathy and myocardial fibrosis were seen in all mice fed component II, but not in those
given component I (Unilever, 1968).
16
Study in pigs
Groups of male and female pigs were given oral doses of Brown FK mixed with a small quantity of
diet immediately before the main feed, at doses of 0, 100, 250, or 500 mg/kg bw/day for 24 weeks.
The principal effect was deposition of pigment in the tissues, described as lipofuscin. This was seen
primarily in the liver of both sexes at all dosage levels, and was accompanied by increased lysosomal
enzyme activity (more marked at the higher dose levels). Pigment was also found in the heart in male
animals, and here it was associated with an increased acid phosphatase activity. Pigment was also
present in the kidneys at all dose levels in males and only at the highest dose level in females. No
adverse effects were observed regarding growth, food consumption, haematological indices, liver and
kidney function, or organ weights. The authors stated that a no-effect level could not be determined in
this study, without providing further details on the basis for this conclusion (Gaunt et al., 1968).
3.2.3.
Genotoxicity
JECFA describes a single study on the genotoxic potential of Brown FK (JECFA, 1985). In this study,
the mutagenic potency of different Brown FK samples (from different manufacturers) and its
individual components were investigated in Salmonella typhimurium (TA1535, TA1537 and TA1538)
with and without metabolic activation with rat liver S9 (Venitt and Bushell, 1976). Brown FK induced
frame shift mutations in TA1538 in the presence of metabolic activation. Mutagenicity was linearly
concentration-dependent in a range of 0-3 mg/plate, with activities ranging from 22 to 50 times the
spontaneous mutation frequency. One sample of Brown FK was also mutagenic in the absence of
metabolic activation producing a 16-fold increase in mutations at 4 mg/plate.
The individual components I and II (each present at about 18% in Brown FK product) were also
mutagenic in TA1538 after metabolic activation (Venitt and Bushell, 1976). Mutagenicity was linearly
dose-related in a range of 0-1 mol/plate, with slopes of 0.35 mutants/nmol for component I and 1.5
mutants/nmol for component II. Four other constituents (unspecified di- and tri-substituted diamines)
and sulphanilic acid were inactive. The authors concluded that components I and II were largely
responsible for the mutagenic activity of Brown FK (Venitt and Bushell, 1976).
A bacterial mutagenicity study, not reported by JECFA, was that of Haveland-Smith and Combes
(Haveland-Smith and Combes, 1979). This study examined the ability of 25 food dyes, including
Brown FK, to cause DNA damage and bacterial mutations in a bacterial assay system (fluctuation
assay in Salmonella typhimurium using TA1538 and in Escherichia coli WP2 uvrA) with and without
rat liver S9 metabolic activation. Brown FK induced base pair substitutions without metabolic
activation, and base pair substitutions and frameshifts in the presence of metabolic activation.
The Panel noted that no information was available on the purity of the material tested in these studies.
No additional data on the genotoxicity of Brown FK are provided in the SCF (1984) and TemaNord
(2002) evaluations and no new studies on the genotoxicity of Brown FK have been published since the
previous evaluations.
The Panel noted the absence of any in vivo genotoxicity data on Brown FK, or in vitro data other than
the bacterial mutagenicity studies described above. The Panel also noted that azo-reduction of Brown
FK may produce sulphonated as well as unsulphonated aromatic amines. Jung et al., (1992) have
reviewed the genotoxicity data of a range of sulphonated aromatic amines. To provide insight in the
effect of sulphonation on the genotoxic potential of phenyl- and naphthylamines, the genotoxicity of
sulphonated aromatic amines was compared with their unsulphonated analogues. It was found that in
general sulphonated phenyl- and napthylamines, including the azo-reduction products of Brown FK
are non-mutagenic to Salmonella typhimurium in Ames tests. For some other sulphonated aromatic
amines, no genotoxicity was also demonstrated with a variety of other test systems in vitro and in vivo
(no details given). Based on the available data, the authors concluded that sulphonated aromatic
17
amines, in contrast with their unsulphonated analogues, have no or very low genotoxic potential.
Hence, the authors concluded that exposure to sulphonated aromatic amines, derived from metabolic
cleavage or present as contaminants in colourings, are unlikely to induce any significant genotoxic
risk. The Panel considered that this conclusion is also applicable to the sulphonated azo-reduction
products of Brown FK. The Panel noted, however, that azo-reduction of Brown FK also produces nonsulphonated aromatic amines (see Figure 2) and that the colour may also contain impurities such as mphenylenediamine and 4-methyl-m-phenylenediamine, the genotoxicity of which could be of concern.
The Panel additionally noted that 4-methyl-m-phenylenediamine (toluene-2,4-diamine) has been
assessed by the International Agency for Research into Cancer (IARC) as a Group 2B carcinogen
(possibly carcinogenic to humans).
3.2.4.
Chronic toxicity and carcinogenicity
Groups of rats (Colworth-Wistar; 32 males/36 females/group) were fed for two years with Brown FK
containing 54.2% coloured components (lower than current specifications and no details available on
component proportions), at levels of 0, 0.01, 0.03, 0.06, 0.1, or 0.5% in the diet (equivalent to 0, 5, 15,
30, 50, or 250 mg/kg bw/day). At dietary levels providing 30 mg Brown FK/kg bw/day and above,
pigment deposition in tissues was seen. At 250 mg/kg bw/day increases in splenic weight and hepatic
granulomata were observed. The authors concluded that the no-effect level was 15 mg/kg bw/day
based on pigment deposition or 30 mg Brown FK/kg bw/day based on evidence of toxicity (Unilever,
1971d). The Panel was unable to confirm these conclusions since it did not have access to the study
report.
In a carcinogenicity study in CD rats (60/sex/group) involving an in utero exposure phase followed by
a 2-year exposure of the offspring, animals received Brown FK in the diet (no detail on component
proportions) at dose levels of 0, 15, 50, or 250 mg/kg bw/day (Amyes et al., 1983; Unilever, 1980;
1983a). Two groups served as untreated controls, and one group received sodium chloride equivalent
to the highest Brown FK-dose group (salt control). The study was terminated at weeks 105 and 110
after weaning of the male and female offspring, respectively.
In this study at the highest dose of 250 mg/kg bw/day, a range of treatment-related effects was
reported, consisting of decreased body-weight gains (14 and 10% decrease compared to controls for
males and females respectively at termination), whereas food consumption was not affected compared
to controls. Clinical chemistry investigations in female rats revealed higher creatine phosphokinase,
lactate dehydrogenase, and hydroxybutyrate dehydrogenase activities compared with untreated
controls, but not compared with salt controls. In male animals, plasma albumin and thyroxine (T4)
concentrations were significantly elevated. In both sexes, isocitrate dehydrogenase activities were
found to be higher. Histopathologically at the highest dose level of 250 mg/kg bw/day, the incidence
of dark thyroid glands was increased and deposition of brown pigment was noted (heart, skeletal
muscle, diaphragm, tongue, thyroid, caecum, and hepatic Kupffer cells), but was not associated with
any tissue reaction. In females, Brown FK treatment was associated with an increased incidence of
pelvic nephrocalcinosis and cystic distension of the follicles of the thyroid. No treatment-related
effects were seen regarding mortality, absolute or relative organ weights, ophthalmoscopy,
haematology, or urinalysis. Frequency and time of onset of palpable swellings were unaffected by
treatment and no increase in tumour incidence compared with controls was seen at any site.
Based on these findings the authors concluded that Brown FK was not carcinogenic in rats, and that
the no-effect level (including for pigment deposition) was 50 mg/kg bw/day (Amyes et al., 1983;
Unilever, 1980; 1983a). The Panel noted however that according to JECFA, histopathology was not
carried out on tissues from animals in the 15 or 50 mg/kg bw/day dose groups (JECFA, 1985, 1986),
and that JECFA had requested a complete histopathological examination of the animals in these dose
groups, in order to derive a NOAEL from this study. The Panel considered, in line with the view of
JECFA, that since treatment-related changes in a range of organs had been reported in the top-dose
group, histopathological examination of the same organs and tissues of the low- and mid-dose group
18
should have been carried out. Since the Panel did not have access to the report of this study, it was
unable to confirm the authors conclusion that the no-effect level in the study (including for pigment
deposition) was 50 mg/kg bw/day.
Groups of mice (Colworth C57Bl, 40/sex/group) were fed Brown FK containing 62.5% coloured
components (no detail on component proportions) at dietary levels of 0, 0.0125, 0.0375, 0.075, 0.125,
or 0.625% (equivalent to approximately 0, 20, 55, 110, 180, or 895 mg/kg bw/day) for 80 weeks. From
a dose of 55 mg/kg bw/day onwards, pigment deposition was observed (not further specified). From a
dose of 110 mg/kg bw/day onwards, the number of animals with hepatic nodules was increased (26,
23, 27, 56, 42 and 64 at the dose levels indicated in the study). Further, at 180 mg/kg bw/day and
above, heart weights were increased, and finally, at the 895 mg/kg bw/day dose level, the effects
consisted of reduced growth and food utilization, increased weights of liver, kidney, spleen, brain and
testes, splenic haemopoiesis and increased myocardial fibrosis, and increased mortality among
females. The incidence of hepatocellular carcinoma was 3, 2, 0, 5, 6 and 2 at the dose levels indicated
in the study (JECFA, 1977; 1985 - quoting Unilever, 1970). The Panel noted that the sex of the mice
showing hepatocellular carcinoma was not specified in the JECFA reports, and did not have access to
the original Unilever report in order to clarify this.
As also described in the section on subchronic toxicity, a number of chronic studies on Brown FK in
rats and mice have focussed on identification of the pigment deposited in tissues following
administration of the dyestuff (e.g. Unilever, 1971a; 1983b). These investigations led to the conclusion
that the deposited pigment was different in characteristics and distribution to lipofuscin, which is a
characteristic degenerative pigment seen in the tissues of older rodents. It has been postulated
therefore that the pigment may not represent evidence of sub-lethal cell damage, but is, instead, an
insoluble oxidation product of a metabolite of the colour. 1,2,4-triaminobenzene (metabolite A) was
shown to be very rapidly oxidized to 1,4,7-triaminophenazine, which is a brown, water-insoluble,
material (Muller, 1889).
No additional data on chronic toxicity and carcinogenicity are provided in the SCF (1984) and
TemaNord (2002) evaluations, and no new studies have been published since the previous evaluations.
3.2.5.
Reproductive and developmental toxicity
In a multi-generation reproduction study, groups of 24 male and 24 female rats were given Brown FK
in the diet (no detail on component proportions) at a level of 300 mg/kg diet (equivalent to 15 mg/kg
bw/day) for five weeks post-weaning and subsequently 600 mg/kg diet (equivalent to 30 mg/kg
bw/day), for three successive generations. Controls received stock diets. Fertility, in utero mortality,
number of young per litter, birth weights, growth rates, gestation indices, viability indices and
lactation indices were unaffected by treatment. At autopsy and histopathological examination no
treatment-related lesions were observed in F3 weanlings. Examination of haematology, clinical
chemistry and organ weights, revealed occasional statistically-significant differences but these were
not consistent and, in the absence of pathological lesions, were not considered to be of toxicological
significance. The researchers concluded that at 30 mg/kg bw/day, Brown FK had no effect on
reproductive performance (Unilever, 1983c).
In a developmental toxicity study, groups of 35 pregnant rats were fed Brown FK at dietary
concentrations of 0, 0.03, 0.15, or 0.6 % (equivalent to 0, 15, 75, or 300 mg/kg bw/day) from day 0-19
post coitus. Additional groups received 0.6% sodium chloride (salt control) or aspirin (250 mg/kg
bw/day) (positive control). Five animals at each dose level were allowed to litter normally and the
offspring were raised to weaning, while the remaining animals in each group were sacrificed at
Gestation Day (GD) 17 and the fetuses removed by Caesarean section. Two-thirds of the fetuses were
examined for gross soft-tissue abnormalities, then cleared and stained with Alizarin red for
examination for skeletal defects. The remaining one-third of animals was examined for soft tissue
19
defects. Soft tissue abnormalities or skeletal defects were not observed in any of the sacrificed fetuses
or weaned animals (Unilever, 1978).
In the 2-year carcinogenicity study in rats with an in utero exposure phase, followed by a 2-year
exposure of the offspring described in section 3.2.4 in which animals received Brown FK in the diet at
dose levels of 0, 15, 50, or 250 mg/kg bw/day, no treatment-related effects were noted regarding
general condition, food intake, body-weight gain, mating performance, conception rate, length of
gestation, litter size or growth and viability of offspring (Amyes et al., 1983; Unilever, 1980; 1983c).
No additional data on reproductive toxicity of Brown FK are provided in the SCF (1984) evaluation
and in TemaNord (2002), and no new studies have been published since the previous evaluations.
3.2.6.
Allergenicity, hypersensitivity and intolerance
Reactions to food colourings, including those triggered by immune (immediate and delayed type
hypersensitivity) and non-immune (intolerance) mechanisms are assumed to be infrequent in the
population, and prevalence of 0.14 to around 2% has been reported (Young et al., 1987; Hannuksela
and Haahtela, 1987; Fuglsang et al., 1993; Fuglsang et al., 1994).
of exposure the incidence is very low if any. On the other hand, the low/absent reports of adverse
4.
Discussion
The Panel was not provided with a newly submitted dossier and based its evaluation on previous
evaluations, additional literature that became available since then and the data available following a
public call for data. The Panel noted that not all original studies, on which previous evaluations were
based, were available for re-evaluation by the Panel.
Brown FK (E 154) is a multi-component mixture of mono-, di- and tri-azo dyes authorised for use as a
food additive in the EU, which has been previously evaluated by both the SCF in1984 and JECFA in
1977 and in 1985.
Specifications have been defined in Commission Directive 2008/128/EC and by JECFA (2006).
4,4-(2,4-diamino-1,3-phenylenebisazo)dibenzenesulphonate (component IV), disodium 4,4-(2,4diamino-5-methyl-1,3-phenylenebisazo)dibenzenesulphonate (component V) and trisodium 4,4,4(2,4-diaminobenzene-1,3,5-trisazo)tribenzenesulphonate (component VI), together with subsidiary
colouring matters, water, sodium chloride and/or sodium sulphate as the principal uncoloured
components. Brown FK is described as the sodium salt. The calcium and the potassium salts are also
permitted (Commission Directive 2008/128/EC).
The purity is specified as not less than 70% total colouring matters. Of the total colouring matters
present, the specifications stipulate that the proportions of the components of the mixture shall not
exceed 26% for component I, 17% each for components II and III, 16% for component IV, 20% for
component V and 16% for component VI. The remaining 30% may be accounted for by sodium
chloride or sodium sulphate, although this is never mentioned explicitly, together with < 3.5%
subsidiary colouring matters, 0.7% 4-aminobenzene-1-sulfonic acid, 0.35% m-phenylenediamine
20
and 4-methyl-m-phenylenediamine, 0.0007% unsulphonated primary aromatic amines other than mphenylenediamine and 4-methyl-m-phenylenediamine and 0.2% water insoluble material.
The Panel noted that according to the CAS Registry Number, Brown FK is comprised of only
components II and IV rather than the six components mentioned by Commission Directive
2008/128/EC. The Panel also noted that, as reported by Emerton (2008), manufacture of Brown FK
according to the EC specifications is extremely difficult and, given the limited usage of the colour and
the availability of alternative colours, the major manufacturers have stopped offering the colour to EC
specifications. The Panel noted therefore that there is some doubt about whether Brown FK is still
used as a food colour in the European Union.
The SCF in its 1984 evaluation of Brown FK derived an ADI of 0.15 mg/kg bw/day based on a
NOAEL of 15 mg/kg bw/day, determined in a rat multi-generation study. The Panel surmised that the
study used was likely to be the study reported by Unilever (Unilever, 1983c) and that the SCF had
used the initial dose level of 15 mg/kg bw/day used in the study, with application of an uncertainty
factor of 100 to arrive at the ADI of 0.15 mg/kg bw/day, although the study authors had reported a
NOAEL of 30 mg/kg bw/day. The latter was the dose level used in the study following weaning of the
F1 generation at week 5.
JECFA (1985, 1986) allocated a temporary ADI of 0.075 mg/kg bw/day to Brown FK based on a
NOAEL of 15 mg/kg bw/day for pigment accumulation in tissues, derived from a 2-year chronic
toxicity study in rats carried out by Unilever. The Panel noted that an uncertainty factor of 200 was
applied to arrive at the ADI of 0.075 mg/kg bw/day. The ADI was made temporary pending results
from the ongoing long-term carcinogenicity study in CD rats with an in utero exposure phase carried
out at dose levels of 0, 15, 50 or 250 mg/kg bw/day and reported by Unilever in 1980 and 1983. In this
study, while a range of treatment-related effects was reported at the highest dose of 250 mg/kg
bw/day, the authors had concluded that the no-effect level (including for pigment deposition) was 50
mg/kg bw/day. However histopathology had not been carried out on organs and tissues showing
treatment-related histopathological lesions in the top-dose group from animals in the low and
intermediate dose groups. JECFA therefore requested a complete histopathological examination of
these animals, in order to establish a definitive NOAEL from this study. At the time of the last
evaluation of Brown FK by JECFA in 1987, the requested data had not been made available and the
temporary ADI was withdrawn (JECFA, 1987).
The available studies on the absorption, distribution, metabolism and excretion of Brown FK indicate
that the individual components of Brown FK are not absorbed as intact molecules to any extent as
such but undergo reductive cleavage in the intestine, with some subsequent absorption of aromatic
polyamine metabolic products which are further acetylated before urinary excretion. Metabolites of
Brown FK, including sulphanilic acid, are also excreted in the faeces. Overall given the multicomponent nature of Brown FK, the metabolism of the mixture is complex but the absorption of
metabolites of Brown FK, including aromatic amines, was demonstrated, as evidenced by the toxicity
seen in specific studies with these metabolites.
Adverse effects seen after short-term or prolonged exposure to Brown FK in rats and/or mice include
damage to cardiac (and to a minor extent skeletal) muscle fibres, characterised by vacuolar myopathy,
degenerative lesions, necrosis and pigment deposition. Other changes induced by Brown FK include
occasional hydropic degeneration of the kidney and fatty change of the liver, an undefined pigment
deposition in various tissues, and increases in organ weight (spleen, liver, heart, testes and thyroid).
The available studies provide no clear indication of a NOAEL for the cardiac toxicity or other
pathological lesions seen following oral administration of Brown FK, other than that this NOAEL
appears to lie below 100 mg/kg bw/day.
In addition to the toxicity findings, pigmentation of tissues was seen from a dose of 55 mg/kg bw/day
onwards in mice and 30 mg/kg bw/day onwards in rats. Regarding tissue pigmentation, an attempt to
differentiate between lipofuscin and Brown FK-induced pigment gave inconclusive results. It has
21
however been proposed that the pigment noted is 1,4,7-triaminophenazine, which is a brown waterinsoluble material. This would indicate that the pigment does not represent evidence of sub-lethal cell
damage, but is, instead, an insoluble oxidation product of a Brown FK metabolite. The lowest NOAEL
reported from a dietary study in rats of 15 mg/kg bw/day was based on pigment deposition rather than
biochemical or pathological indicators of toxicity.
In an in vitro bacterial mutagenicity study, different samples of Brown FK have been shown to induce
concentration-dependent frame shift mutations (22- to 50-times the spontaneous mutation frequency)
in Salmonella typhimurium (TA1538). The genotoxicity of samples of Brown FK has also been
demonstrated in fluctuation assays in Salmonella typhimurium using TA1538 and in Escherichia coli
WP2 uvrA, with and without rat liver S9 metabolic activation. The Panel noted that the different
components of Brown FK are metabolised to non-sulphonated as well as sulphonated aromatic amines,
and that potential impurities in the commercial product include m-phenylenediamine and 4-methyl-mphenylenediamine and other non-specified non-sulphonated aromatic amines, the genotoxicity of
which could be of concern. The Panel additionally noted that 4-methyl-m-phenylenediamine (toluene2,4-diamine) has been assessed by the IARC as a Group 2B carcinogen (possibly carcinogenic to
humans). While the Panel considered that the positive results obtained in in vitro bacterial
mutagenicity studies on Brown FK, together with the potential genotoxicity of some of its metabolites
and possible impurities, give rise to some concern regarding the genotoxicity of Brown FK, this
concern is allayed by the results of long-term studies in rats and mice showing no convincing evidence
of carcinogenicity of Brown FK.
Three long-term toxicological studies have been conducted, one in mice and two in rats. One of the rat
studies was described as a 2-year chronic toxicity study, while the other rat study was a
carcinogenicity study with in utero exposure followed by a 2-year exposure of the offspring. In the
mouse study, which was an 80-week carcinogenicity study, hepatocellular carcinomas were observed.
However the Panel considered that since the relevance of hepatic tumours in mice for humans has
been questioned (e.g. Gold and Slone, 1995), the results of this study are unlikely to be of biological
relevance and are therefore not a suitable basis for risk assessment. No indications for carcinogenicity
were obtained in the long-term study with rats involving in utero exposure. The Panel concluded
therefore that these long-term studies of Brown FK in rats and mice show no evidence of carcinogenic
effects relevant to humans.
No evidence of reproductive toxicity of Brown FK was seen in a multi-generation study and a
developmental toxicity study in rats. The NOAEL of 15 mg/kg bw/day in the multi-generation study,
(corresponding to the dose level of 300 mg Brown FK/kg diet used at the commencement of the study)
is likely to have been the point of departure used by the SCF in 1984 to derive an ADI of 0.15 mg/kg
bw/day for Brown FK.
The Panel noted that, in addition to the multi-generation study in rats used by SCF to derive an ADI,
either the 2-year chronic toxicity study in rats with a reported NOAEL of 15 mg Brown FK/kg bw/day
(used by JECFA to derive their temporary ADI of 0.075 mg/kg bw/day for Brown FK) or the 2-year
carcinogenicity study in rats involving an in utero exposure with a reported (by the authors) NOAEL
of 50 mg/kg bw/day, might be considered to be appropriate studies to use as the basis for establishing
an ADI.
The Panel did not however have access to the reports of any of these studies, and noted the
uncertainties surrounding the NOAELs derived by the authors of the studies, including the relative
contributions of treatment-related changes such as myopathy, myocardial fibrosis and pigment
deposition in establishing effect and no-effect levels. The Panel noted that the reported NOAEL in the
2-year chronic toxicity study in rats was 15 mg Brown FK/kg bw/day, while in the 2-year
carcinogenicity study in rats involving an in utero exposure it was reported to be 50 mg Brown FK/kg
bw/day. In particular the Panel noted that according to JECFA, a range of treatment-related effects
were reported in the latter study at the highest dose of 250 mg/kg bw/day. However histopathological
22
examination of the organs and tissues showing treatment-related changes in the highest dose group had
not been carried out in the animals in the low and intermediate dose groups.
Since treatment-related effects had been observed at the top dose level of 250 mg/kg bw/day in this
study, the Panel considered, in agreement with the view of JECFA, that histopathological examination
of the relevant organs and tissues of (at least) the intermediate dose group animals should have been
carried out. The Panel concluded that given the uncertainties in the available data sets, it was not
possible to derive an appropriate NOAEL from the available data on these studies that could be used
to establish an ADI. Therefore, the Panel is unable to conclude on the safety of Brown FK.
of exposure the incidence is very low if any. On the other hand, the low/absent reports of adverse
Based on the fact that Brown FK is permitted only in kippers at a maximum concentration of 20
mg/kg, the Panel calculated an anticipated refined dietary exposure (Tier 2) for the European
population ranging from 0.03 (in average) up to 0.14 mg/kg bw/day at the 95th percentile in children
(aged 1-10 years old and weighing 15-30 kg) and ranging from 0.002 (in average) up to 0.14 mg/kg
bw/day at the 95th percentile in adults (aged >18 years old and weighing 60 kg).
The Panel notes that the JECFA specification for lead in Brown FK is < 2 mg/kg, whereas the EC
The Panel also notes that the aluminium lake of the colour could add to the daily intake of aluminium,
for which a TWI of 1 mg aluminium/kg bw/week has been established, and that therefore
specifications for the maximum level of aluminium in the lakes may be required.
CONCLUSIONS
Brown FK (E 154) is a multi-component mixture of mono-, di- and tri-azo dyes authorised as a food
additive in the EU, which has been previously evaluated by both the SCF in 1984 and JECFA in 1977
and 1985.
The SCF in its evaluation of Brown FK in 1984 established an ADI of 0.15 mg/kg bw based on a
NOAEL of 15 mg/kg bw/day, determined in a rat multi-generation study, while JECFA allocated a
temporary ADI of 0.075 mg/kg bw/day for Brown FK based on pigment accumulation in tissues with
a NOAEL of 15 mg/kg bw/day in a 2-year chronic toxicity study in rats. The ADI was made
temporary pending further results from a 2-year carcinogenicity study in CD rats with an in utero
exposure phase. In this study, while a range of treatment-related effects were reported at the highest
dose of 250 mg/kg bw/day, the authors had concluded that the no-effect level (including for pigment
deposition) in this study was 50 mg/kg bw/day. JECFA noted however that histopathology had not
been carried out on tissues from animals receiving 15 or 50 mg/kg bw/day. JECFA therefore requested
a complete histopathological examination of these animals in order to confirm the NOAEL from the
study. At the time of the last evaluation of Brown FK by JECFA in 1987, the requested data had not
been provided and the temporary ADI of 0.075 mg/kg bw/day for Brown FK was withdrawn.
The Panel concludes that there are considerable uncertainties surrounding the NOAELs derived by the
authors of any of these studies, including the relative contributions of treatment-related changes such
as myopathy, myocardial fibrosis and pigment deposition in establishing effect and no-effect levels.
The Panel did not have access to the reports of any of these studies and could not confirm the reported
findings. In particular the Panel noted that according to JECFA, while a range of treatment-related
effects were reported in the 2-year carcinogenicity study in rats with an in utero exposure phase at the
23
highest dose of 250 mg/kg bw/day, histopathological examination of the animals in the low and
intermediate dose groups had not been carried out. Since treatment-related effects had been observed
at the top dose level of 250 mg/kg bw/day in this study, the Panel considered, in line with the view of
JECFA, that histopathological examination of the relevant organs and tissues of (at least) the
intermediate dose group animals should have been carried out.
The Panel therefore concludes that based on the uncertainties in the available data sets, it is not
possible to derive an appropriate NOAEL from these studies that could be used to establish an ADI.
Therefore the Panel is unable to conclude on the safety of Brown FK given the deficiencies in the
available toxicity database on the colour.
Based on the fact that Brown FK is permitted in kippers only, at a maximum concentration of 20
mg/kg, the Panel calculated an anticipated refined dietary exposure (Tier 2), for the European
population ranging from 0.03 (in average) up to 0.14 mg/kg bw/day at the 95th percentile in children
(aged 1-10 years old and weighing 15-30 kg) and ranging from 0.002 (in average) up to 0.14 mg/kg
bw/day at the 95th percentile in adults (aged >18 years old and weighing 60 kg).
The Panel notes that no cases of intolerance/allergenicity have been reported after oral exposure to
Brown FK and that no data on sensitivity to Brown FK are available.
The Panel notes that the JECFA specification for lead in Brown FK is < 2 mg/kg whereas the EC
The Panel noted that the aluminium lake of the colour could add to the daily intake of aluminium for
which a TWI of 1 mg aluminium/kg bw/week has been established and that therefore specifications
for the maximum level of aluminium in the lakes may be required.
DOCUMENTATION PROVIDED TO EFSA

1.
Pre-evaluation document prepared by the Dutch National Institute for Public Health and the
Environment (RIVM), Bilthoven, The Netherlands.
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28
GLOSSARY AND ABBREVIATIONS

ADI
Acceptable Daily Intake
ANS
Scientific Panel on Food Additives and Nutrient Sources added to 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
BIBRA
British Industrial Biological Research Association
AST
Aspartate aminotransferase
CAS
Chemical Abstracts Service
EC
European Commission
EFSA
European Food Safety Authority
EU
European Union
EXPOCHI
Individual food consumption data and exposure assessment studies for children
FAO/WHO
Food and Agriculture Organization/World Health Organization
GD
Gestation Day
GLP
Good Laboratory Practice
HCl
Hydrochloric acid
HPLC
High-performance liquid chromatography
IARC
International Agency for Research into Cancer
JECFA
Joint FAO/WHO Expert Committee on Food Additives
NOAEL
No-Observable-Adverse-Effect-Level
SCF
Scientific Committee on Food
TWI
Tolerable Weekly Intake
UK NDNS
UK Detailed individual food consumption data (National Diet and Nutrition

Survey)
UNESDA
Union of European Beverage Associations
29

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EFSA Journal 2010;8(4):1535

Scientific Opinion on the re-evaluation of Brown FK (E 154) as a food

European Food Safety Authority, 2010

Re-evaluation of Brown FK (E 154) as a food additive

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

No data on sensitivity to Brown FK are available. Additionally, no cases of intolerance/allergenicity

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

BACKGROUND AS PROVIDED BY THE EUROPEAN COMMISSION

TERMS OF REFERENCE AS PROVIDED BY THE EUROPEAN COMMISSION

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

Identity of the substance

sodium 4-(2,4-diaminophenylazo)benzenesulphonate - C12H11N4NaO3S

Brown FK is soluble in water and sparingly soluble in ethanol (JECFA, 2006).

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

Re-evaluation of Brown FK (E 154) as a food additive

Specifications for Brown FK according to Commission Directive 2008/128/EC and

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

Methods of analysis in foods

Reaction and fate in food

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

Case of need and proposed uses

Information on existing authorisations and evaluations

Brown FK is permitted as a food additive in the EU under Directive 94/36/EC.

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

Table 2: Summary of anticipated exposure to Brown FK in children and adult populations

Biological and toxicological data

Absorption, distribution, metabolism and excretion

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

Acute oral toxicity

Short-term and subchronic toxicity

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

Re-evaluation of Brown FK (E 154) as a food additive

Chronic toxicity and carcinogenicity

Re-evaluation of Brown FK (E 154) as a food additive

Reproductive and developmental toxicity

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

Allergenicity, hypersensitivity and intolerance

Re-evaluation of Brown FK (E 154) as a food additive

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive

EFSA Journal 2010;8(4):1535

Re-evaluation of Brown FK (E 154) as a food additive