Diammonio(ethyl)[4-[[4-[ethyl(3-sulphonatobenzyl)amino]phenyl](2-sulphonatophenyl)methylene]cyclohexa-2,5-dien-1-ylidene](3-sulphonatobenzyl)ammonium

Endpoint summary

Description of key information

NOAEL (repeated oral toxicity, chronic study, male rat) = 1072 mg/kg bw/ day

NOAEL (repeated oral toxicity, chronic study, female rat) = 631 mg/kg bw/ day

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

631 mg/kg bw/day

Study duration:

chronic

Species:

rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Mode of Action Analysis / Human Relevance Framework

Toxicokinetic studies suggest that the substance is poorly taken up after ingestion. The colorant has a high molecular weight and is highly soluble in water.

Lack of significant uptake is consistent with the absence of a long-term human health hazard. High doses are osmolytically active which in oral dosing settings may result in diarrhea.

Additional information

The following reported data were obtained for Similar Substance 1. It is expected that the Target substance will present similar effects of repeated dose toxicity. Justification for Read Across is given in Section 13 of IUCLID.

For this dossier, the evaluation of the effects of Similar Substance 1 is also based on the SCCS opinion by EFSA (2010) and on the published data used therein. In EFSA opinion, evaluators had the full study reports from which the literature publications were written, thus, the data reported are well documented and derives from the full source.

 

The key study for chronic hazard assessment is a life-time feeding study in rats with dietary concentrations of 0.1, 1 and 2% (w/w) published by Borzelleca et al (1990). The study design includes the requirements of OECD testing guideline 453, and includes additional fertility and developmental toxicity elements. The chronic toxicity study started with in-utero exposure. Parental animals (each 60 males and females) were exposed via the diet during pre-mating, mating and pregnancy for a total period of 2 months. Then the offspring (70 males and 70 females) was weaned onto the test-material containing diet and kept on this diet for life-long exposure (30 months). For each ten male and female rats, an interim sacrifice was performed after 12 months. Calculated from the body weights, the doses for males were 0, 50, 514, 1072 mg/kg bw and those for females were 0, 62, 631, 1319 mg/kg bw.

 During the study, there was blue staining of fur, feces and exposed skin which is caused by the blue color of the test substance. There were no adverse observations regarding clinical signs, mortality, urinalysis, haematology, clinical chemistry, ophthalmoscopy, organ weights and histopathology. High dose animals showed a slightly higher food consumption. After week 90, mean body weights of the 2.0% females began a steady downward trend that was statistically significant (P < 0.01) from week 102 until the end of the study; terminal body weights were 15% lower compared to control group animals. High dose females showed a reduced survival rate (10/70 versus 28/70 and 24/70 in two control groups) compared to the control group. A NOAEL of 631 mg/kg bw for female rats and of 1072 mg/kg bw for male rats were thus considered.

The same authors also published the results of a life-long feeding study in mice with 0.5, 1.5, and 5% (w/w) in the diet, but this study only included parameters related to carcinogenic properties. No adverse findings were noted in that study. Calculated for daily uptake per kg body weight, doses were 0, 661, 2064, 7354 mg/kg bw for males and 0, 819, 2562, 8966 mg/kg bw for females.

 

Another study has been reported in which Similar Substance 1 caused severe growth retardation in Wistar rats when added to a low fiber diet at a level of 5% for 21 days (Tsujita,1979). The authors showed that concurrent addition of dietary fiber from the roots of edible burdock completely protected against this toxic effect. That study was performed with five male rats per group and only investigated a few parameters. The growth retardation is considered to be the result of malnutrition as the ionic and non-absorbable substance has osmolytic activity and caused chronic diarrhea. The dose of 5% in the diet is excessive and above the limit doses for regulatory testing.

 

Limited investigations with beagle dogs were published by Hansen (1966). Five and three beagle dogs received a dietary level of 2% and 1% for one year, respectively. One dog at the top dose died after 17 days and another died after 46 weeks at the low dose, as a result of - in the view of the authors - intercurrent virus infections. No clinical signs, gross lesions or histological abnormalities observed were observed (Hansen,1966). The doses correspond to ca 500 and 250 mg/kg bw per day.

In conclusion, Similar Substance 1 has been investigated in several studies, most of them being done prior to the introduction of GLP and harmonized testing guidelines. The European Food Safety Agency (EFSA) re-evaluated all available experimental data in 2010 (EFSA Journal 2010) and for the use as a colorant of food stuff issued an acceptable daily intake of 6 mg/kg bw derived from a NOEL of 631 mg/kg bw reported in a chronic feeding study in rats (Borzelleca, 1990).

 

The high molecular weight and the octanol-water-partition coefficient indicate that the substance is unlikely to penetrate the skin.

 

An assessment of the inhalation route is not performed as the substance is not volatile and handled in a non-inhalable form (granules / water-based solutions).

 

References:

- Borzelleca, J.F. et al. 1990.Lifetime Toxicity/Carcinogenicity Studies of FD&C Blue no. 1 (Brilliant Blue FCF) in Rats and Mice.Fd. Chem. Toxicol.28, 221-234.

- EFSA 2010. Scientific Opinion on the re-evaluation of Brilliant Blue FCF (E 133) as a food additive1,EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS).

- Hansen et al.,1966. Chronic Toxicity of Two Food Colors, Brilliant BIue FCF and Indigotine.Toxicol. Appl. Pharmacol.8, 29-36.

- Tsujita et al., 1979. Comparison of protective activity of dietary fibre against the toxicities of various food colours in rats. Nutr. Rep. Intern.20, 635-642.

 

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), the alert value for classification in Category 2 is 100 mg/kg bw/day for repeated oral toxicity in rat. A NOEL of 631 mg/kg bw is reported in a chronic feeding study in rats (Borzelleca, 1990). This value is higher than the alert one. As a consequence, it can be conlcuded that the substance requires no classification for repeated oral toxicity.