Scientific Opinion on the re-evaluation of Amaranth (E 123) as a food additive

The Panel on Food Additives and Nutrient Sources added to Food provides a scientific opinion re-evaluating the safety of Amaranth (E 123). Amaranth has been previously evaluated by JECFA in 1972, 1975, 1978 and 1984, and the SCF in 1976, 1979 and 1983. In 1984 the SCF set an ADI for Amaranth of 0-0.8 mg/kg bw/day based on results from a 90-day rat study. In contrast, in 1984 JECFA allocated an ADI of 0-0.5 mg/kg bw/day Amaranth based on a long-term carcinogenicity study in rats. In evaluating the overall toxicological database on Amaranth, the Panel concludes that the point of departure for establishing an ADI for Amaranth can be defined as 15 mg/kg bw/day, taking both the results from the 2-year study and the reproductive and developmental toxicity studies into account. Therefore using an uncertainty factor of 100, the Panel establishes an ADI for Amaranth of 0.15 mg/kg bw/day. The Panel concludes that at the maximum permitted level of use and/or reported use levels of Amaranth (Tier 2), estimates of anticipated exposure for children (1-14 years old) are around 30 times lower than the ADI of 0.15 mg/kg bw/day at the high percentiles (95th/97.5th). However, for adults the anticipated exposure to Amaranth at the 97.5th percentile can be up to 6 times higher than the ADI. The Panel also notes that main contributors to total anticipated exposure to Amaranth for adults were from aperitif wine drinks and Americano. The Panel notes that anticipated exposure to these uses have been made with the maximum permitted levels of use for Americano although no usage value for this beverage was provided by industry, and with the maximum reported levels of use for aperitif wine drinks, which were reported by Industry to be at the same level as the maximum permitted level.

© European Food Safety Authority, 2010

Following a request from the European Commission, the Panel on Food Additives and Nutrient Sources added to Food (ANS) was asked to deliver a scientific opinion re-evaluating the safety of Amaranth when used as a food colouring substance.

Amaranth (E 123) is an azo dye authorised as a food additive in the EU and has previously been evaluated by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) in 1972, 1975, 1978 and 1984, and by the EU Scientific Committee for Food (SCF) in 1976, 1979 and 1983. JECFA and the SCF established different Acceptable Daily Intakes (ADIs) of 0-0.5 and 0-0.8 mg/kg bw/day Amaranth, respectively.

The Panel was not provided with a newly submitted dossier and based its evaluation on previous evaluations and reviews, additional literature that has become available since then and the data available following a public call for data. The Panel notes that not all original studies on which previous evaluations or reviews were based were available for re-evaluation by the Panel.

The Panel concurs with the view expressed in previous evaluations by JECFA and TemaNord that the absorption of Amaranth is limited, but that after azo-reduction in the gastrointestinal tract, free sulphonated aromatic amines may reach the systemic circulation.

The Panel evaluated the available data on genotoxicity including an in vivo Comet assay in which Amaranth induced significant increases in migration of nuclear DNA in both glandular stomach and the colon of male mice. The Panel considers, in light of the negative carcinogenicity studies and negative results in standard in vivo genotoxicity studies, that the biological significance of the positive genotoxicity results is uncertain. Therefore, the Panel concludes that the effects of Amaranth reported in these genotoxicity studies are not expected to result in carcinogenicity. Furthermore, the Panel notes that a dominant lethal test in male rats was reported to be negative.

Overall, based on the weight-of-evidence of the available data, the Panel considers, in line with the opinions expressed by the SCF, JECFA and TemaNord, that there is no concern with respect to the genotoxicity of Amaranth.

The conversion of Amaranth by azo-reduction in vivo, results in the formation of sulphonated naphthylamines that may not be formed in the standard in vitro genotoxicity tests. In a review by Jung et al. (1992), a range of sulphonated aromatic amines was shown, in general, not to be associated with genotoxicity in vitro and in vivo. Since all the sulphonated aromatic amine metabolites that could in theory be formed by azo-reduction of Amaranth, including naphthionic acid, were considered in the study, the Panel concludes that the data reviewed by Jung et al. (1992) are sufficiently reassuring to support the conclusion that the sulphonated aromatic amines formed from Amaranth by azo-reduction do not give reason for concern with respect to genotoxicity.

Both JECFA and the SCF concluded that Amaranth is not carcinogenic to rats exposed in utero and then subsequently exposed for more than 2 years at doses up to 1250 mg/kg bw/day. The Panel agrees with this conclusion, taking into consideration other available studies on Amaranth, including the most recent, comprehensive 2-year study in rats carried out by British Industrial Biological Research Association (BIBRA) (Clode et al., 1987).

In the 2-year study in rats carried out by BIBRA, renal calcification and hyperplasia were observed at all doses, and therefore no No-Observed-Adverse-Effect-Level (NOAEL) could be identified for Amaranth (Clode et al., 1987). Therefore, and in addition to the afore-mentioned 28-day/90-day study, a re-evaluation of the histology of the renal tissues from the 2-year study reported by Clode et al. (1987) was carried out by Butler and Conning (1983). In the report of this re-evaluation, in respect of renal calcification and hyperplasia a NOAEL of 50 mg/kg bw/day was identified by the authors for Amaranth. JECFA used this NOAEL to establish an ADI of 0-0.5 mg/kg bw/day. The Panel has re-evaluated this study based on the available data of both sets of histopathological analyses and considers the dose of 50 mg/kg bw/day as a Low-Observed-Adverse-Effect-Level (LOAEL), rather than a NOAEL, for renal pelvic calcification and hyperplasia in female rats. According to the Panel, the treatment-related increase in incidence of these changes in Amaranth-treated rats could be an exacerbation of developing senile nephrosis, as suggested by the authors (Butler and Conning, 1983).

Several studies examined the reproductive or developmental toxicity of Amaranth. Due to methodological insufficiencies, many of them were not conclusive in the determination of any reliable NOAEL in the rat, mouse, hamster and rabbit. Several studies were negative in terms of reproduction toxicity in rats or developmental toxicity in mice, rats, rabbits and dogs. Consequently in these studies, a NOAEL was only considered at the highest dose tested. NOAELs were also determined by considering different endpoints. There have been frequent suggestions of increased resorptions indicating embryotoxicity, but repeating the experiments with improved experimental designs has usually failed to confirm this. Taking all the reproduction and developmental studies into account, NOAELs for Amaranth can be identified in the following species tested: mouse 100 mg/kg bw/day (highest dose tested), rat 15 mg/kg bw/day, rabbit 15 mg/kg bw/day (highest dose tested), cat 50 mg/kg bw/day and dog 75 mg/kg bw/day (approximately).

The Panel concludes that while some sensitivity reactions after Amaranth exposure have been reported, no conclusion on the induction of sensitivity by Amaranth could be drawn from the limited scientific evidence available.

The Panel, in evaluating the overall toxicological database on Amaranth notes that several studies are relevant for establishment of the ADI. Based on these considerations the Panel concludes that the point of departure for establishing an ADI for Amaranth can be defined as 15 mg/kg bw/day, taking both the results from the 2-year study and the reproductive, developmental toxicity studies into account. Therefore using an uncertainty factor of 100, the Panel establishes an ADI for Amaranth of 0.15 mg/kg bw/day.

For children (1-14 years old), Amaranth exposure estimates have been calculated from fish roe consumption for six European countries (Cyprus, Finland, Germany, Greece, UK and Sweden). For the adult population, the Panel has selected the UK population as representative of the EU consumers for Amaranth exposure estimates.

The Panel concludes that at the maximum permitted level of use and/or reported use levels of Amaranth (Tier 2), estimates of anticipated exposure for 1- to 14-year old children are around 30 times lower than the ADI of 0.15 mg/kg bw/day at the high percentiles (95^th/97.5^th). However, for adults the anticipated exposure to Amaranth at the high percentile (97.5^th) can be up to 6 times higher than the ADI.

The Panel also notes that the main contributors to total anticipated exposure for adults were from aperitif wine drinks and Americano. The Panel notes that anticipated exposure to these uses have been made with the maximum permitted level of use for Americano, although no usage value was provided by industry for this beverage, and with the maximum reported levels of use for aperitif wine drinks, which were reported by Industry to be at the same level as the maximum permitted level.

The Panel further notes that the specifications for Amaranth need to be updated with respect to the percentage of material not accounted for that may represent sodium chloride or sodium sulphate as the principal uncoloured components.

The Panel notes that the JECFA specification for lead is ≤ 2 mg/kg whereas the EC specification is ≤ 10 mg/kg.

The Panel notes that the aluminium lake of the colour could add to the daily intake of aluminium for which a Tolerable Weekly Intake of 1 mg/kg bw/week aluminium has been established and that therefore specifications for the maximum level of aluminium in the lakes of Amaranth may be required
 

Amaranth, E 123, CAS Registry Number 915-67-3, trisodium 2-hydroxy-1-(4-sulphonato-1- naphthylazo)naphthalene-3,6-disulphonate, CI Food Red 9, food colouring substance