3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid

Endpoint summary

Description of key information

Repeated dose toxicity: oral

In a repeated dose oral toxicity study, Fischer-344 (F-344) male rats were treated with3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydr oxy-3-oxoxanthen-9-yl) benzoic acid (D&C Red No. 28) (CAS Number: 18472-87-2) orally in diet in the concentration of 500 mg/kg/day. Increase in body weight gain was observed in treated rats. As there is no control in the study the effect were not supposed to be treatment related. Daily intake would be 500 mg/kg for 14 days. Therefore, NOAEL was considered to be 500 mg/kg/day on the basis of body weight when Fischer-344 (F-344) male rats were treated with3,4,5,6-tetrachloro-2-(1,4,5,8 -tetrabromo-6-hydr oxy-3-oxoxanthen-9-yl)benzoic acid (D&C Red No. 28) (CAS Number: 18472-87-2)for 14 days.

Repeated dose toxicity: Inhalation

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. Taking into account the low vapour pressure of the substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid, which is reported as 3.12E-021 Pa. Also considering the particle size distribution of the substance, the majority of the particles were found to be in the size of 75.0 micrometer which is much larger size range compared to the inhalable particulate matter. Thus, exposure to inhalable dust, mist and vapour of the chemical 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2) is highly unlikely. Therefore this study is considered for waiver.

Repeated dose toxicity: Dermal

The acute toxicity value for 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (as provided in section 7.2.3) is >2000 mg/kg body weight. The substance was also found to be not irrtating to skin. Also, given the use of the chemical as dye; repeated exposure by the dermal route is unlikely. Thus, it is expected that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no dermal absorption data as well as no data available that suggests that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from peer reviewed journal

Qualifier:

no guideline available

Principles of method if other than guideline:

Subacute repeated dose toxicity study of D&C Red No. 28 orally in rats was conducted.

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of test material: Acid Red 92
- Molecular formula: C20H2Br4Cl4O5Na2
- Molecular weight: 829.66 g/mol
- Substance type: Organic
- Physical state: Red powder
- Impurities (identity and concentrations):6-0.5 %

Species:

rat

Strain:

Fischer 344

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Sprague–Dawley Inc. (Indianapolis, IN).
- Age at study initiation: 7-weeks-old
- Weight at study initiation: 145g ± 8 g
- Fasting period before study: 12 hour
- Housing: Animals were housed in individual Nalgene metabolism cages or wire hanging cages.
- Diet (e.g. ad libitum): Teklad 4% Mouse-Rat Diet, ad libitum
- Water (e.g. ad libitum): Water, ad libitum
- Acclimation period: 5–7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 23.33 °C
- Humidity (%): 40-60%
- Air changes (per hr): 15 fresh filtered air changes per hour
- Photoperiod (hrs dark / hrs light): 12 h light/dark cycle

IN-LIFE DATES: From: To: No data available

Route of administration:

oral: feed

Vehicle:

other: Blended rat chow (Teklad 4% Mouse-Rat Diet)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: Diet was prepared by blending rat chow (Teklad 4% Mouse-Rat Diet) mixed with Red 28 for 14 days such that their daily intake would be 500 mg/kg.

DIET PREPARATION
- Rate of preparation of diet (frequency): Every three days
- Mixing appropriate amounts with (Type of food): Blending rat chow (Teklad 4% Mouse-Rat Diet)
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): blending rat chow (Teklad 4% Mouse-Rat Diet)
- Concentration in vehicle: 500 mg/kg/day
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

14 days

Frequency of treatment:

Daily

Dose / conc.:

500 mg/kg bw/day (nominal)

No. of animals per sex per dose:

500 mg/kg: 56 male rats

Control animals:

not specified

Details on study design:

No data available

Positive control:

No data available

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: No data available
- Time schedule: No data available
- Cage side observations checked in table [No.?] were included. No data available

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No data available

BODY WEIGHT: Yes
- Time schedule for examinations: Every other day

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY: No data available
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available
- Time schedule for examinations: No data available

OPHTHALMOSCOPIC EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available

HAEMATOLOGY: No data available
- Time schedule for collection of blood: No data available
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined. No data available

CLINICAL CHEMISTRY: No data available
- Time schedule for collection of blood: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined. No data available

URINALYSIS: Yes
- Time schedule for collection of urine: No data available
- Metabolism cages used for collection of urine: No data available
- Animals fasted: Yes, for 24 hours
- Parameters checked in table [No.?] were examined. Presences of D&C Red No. 28 in urine were examined.

NEUROBEHAVIOURAL EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data available

OTHER: No data available

Sacrifice and pathology:

No data available

Other examinations:

No data available

Statistics:

No data available

Clinical signs:

not specified

Mortality:

not specified

Body weight and weight changes:

no effects observed

Description (incidence and severity):

The animals gained weight over the 14-day feeding period on the Red 28 diet. As there is no control in the study the effect were not supposed to be treatment related.

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

no effects observed

Description (incidence and severity):

No dye was detected in the urine or cage rinse of the animals pretreated with Red 28 in the diet.

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

body weight and weight gain

other: No effects was observed

Critical effects observed:

no

Conclusions:

NOAEL was considered to be 500 mg/kg/day when Fischer-344 (F-344) male rats were treated with 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydr oxy-3-oxoxanthen-9-yl)benzoic acid (D&C Red No. 28) (CAS Number: 18472-87-2).

Executive summary:

In a repeated dose oral toxicity study, Fischer-344 (F-344) male rats were treated with 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydr oxy-3-oxoxanthen-9-yl) benzoic acid (D&C Red No. 28) (CAS Number: 18472-87-2) orally in diet in the concentration of 500 mg/kg/day. Increase in body weight gain was observed in treated rats. As there is no control in the study the effect were not supposed to be treatment related. Daily intake would be 500 mg/kg for 14 days. Therefore, NOAEL was considered to be 500 mg/kg/day on the basis of body weight when Fischer-344 (F-344) male rats were treated with 3,4,5,6-tetrachloro-2-(1,4,5,8 -tetrabromo-6-hydr oxy-3-oxoxanthen-9-yl)benzoic acid (D&C Red No. 28) (CAS Number: 18472-87-2) for 14 days.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

500 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The data is Klimicsh 2 and from peer reviewed journal.

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

Additional information

Repeated dose toxicity: oral

Various repeated dose toxicity studies has been investigated to observe the adverse general toxicological effects occurring as a result of repeated daily dosing with, or exposure, to a substance for a specified period up to the expected lifespan of the test species. Often are the studies based on experimental data in rodents for 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2). The studies are summarized as below:

 

In a repeated dose oral toxicity study by C.J. Sweeta et al. (Food and Chemical Toxicology 42 (2004) 641–648), Fischer-344 (F-344) male rats were treated with 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydr oxy-3-oxoxanthen-9-yl) benzoic acid (D&C Red No. 28) (CAS Number: 18472-87-2) orally in diet in the concentration of 500 mg/kg/day. Increase in body weight gain was observed in treated rats. As there is no control in the study the effect were not supposed to be treatment related. Daily intake would be 500 mg/kg for 14 days. Therefore, NOAEL was considered to be 500 mg/kg/day on the basis of body weight when Fischer-344 (F-344) male rats were treated with 3,4,5,6-tetrachloro-2-(1,4,5,8 -tetrabromo-6-hydr oxy-3-oxoxanthen-9-yl)benzoic acid (D&C Red No. 28) (CAS Number: 18472-87-2) for 14 days.

 

 

Further, in repeated dose toxicity study published in a SCCNFP study report (2004), male and female HanBrl: WIST (SPF) rats were exposed to Acid Red 92 in the concentration of 0, 10, 50 and 250 mg/kg bw/day orally for 4 weeks. No effect were observed on survival, food consumption and organ weight of treated rats. Dose related reddish faeces was observed at 10, 50 and 250 mg/kg bw/day. Decreased percentage of basophile and increased platelet count were observed in 250 mg/kg/day treated female rats. Stomach irritation in both male and female rats at 50 and 250 mg/kg bw/day and Focal spongiosis of the limiting ridge in male and female rat and dyskeratosis in female rat at 250 mg/kg/day and Dyskeratosis in female rat were observed at 50 mg/kg/day. Therefore, NOAEL was considered to be 10 mg/kg/day when male and female HanBrl: WIST (SPF) rats were exposed to 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo -6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid Red 92) orally for 4 weeks.

 

Similarly, in repeated dose toxicity study published in a SCCNFP study report (2004), male and female HanBrl: WIST (SPF) rats were exposed to 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy- 3-oxoxanthen -9-yl)benzoic acid (Acid Red 92) (CAS No. 18472-87-2) in the concentration of 0, 10, 50 and 250 mg/kg bw/day orally for 13 weeks. One male and one female rat died in 50 mg/kg bw/day dose group. Deaths are considered to be result of dosing errors. Reddish feces in all dose groups were observed. However, no effects were observed on body weight, food consumption and ophthalmoscopy. Increased absolute eosinophils count in female rat. Decreased triglycerides; protein levels and ALAT activity at 250 mg/kg/day in male rats were observed. Impaired concentrating ability (increased volume and decreased density) in male and female, Increased urinary pH in male and Reddish urine were observed at 250mg/kg bw/day. Reddish urine were observed at 50mg/kg bw/day. Myosisin males and female rats and decreased locomotor activity in male rats were observed at 250mg/kg bw/day. In addition, passive discoloration of various segments of the digestive tract in all 10, 50 and 250 mg/kg bw/day rats and stomach irritation in both sexes at 250 mg/kg bw/day (vacuolation limiting ridge epithelium, hyaline inclusions in glandular mucosa and squamous hyperplasia in most males and females, and submucosal cell infiltrate in all females). In males of the 50 mg/kg bw/day group slight stomach irritation was observed (squamous hyperplasia) in a number of males. Therefore, NOAEL was considered to be 10 mg/kg/day when male and female HanBrl: WIST (SPF) rats were exposed to 3,4,5,6-tetrachloro-2- (1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid Red 92) (CAS No. 18472 -87 -2) orally for 13 weeks.

 

On the basis of evidence from above studies for target substance in experimental animals, it can be presumed that there is no potential to be harmful to human health following repeated exposure. The substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2) is unclassified because no specific target organ toxicity was seen. Thus, on the basis of CLP classification criteria the substance is not classified.

 

Repeated dose toxicity: inhalation

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. Taking into account the low vapour pressure of the substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid, which is reported as 3.12E-021 Pa. Also considering the particle size distribution of the substance, the majority of the particles were found to be in the size of 75.0 micrometer which is much larger size range compared to the inhalable particulate matter. Thus, exposure to inhalable dust, mist and vapour of the chemical 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2) is highly unlikely. Therefore this study is considered for waiver.

 

Repeated dose toxicity: dermal

The acute toxicity value for 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (as provided in section 7.2.3) is >2000 mg/kg body weight. The substance was also found to be not irrtating to skin. Also, given the use of the chemical as dye; repeated exposure by the dermal route is unlikely. Thus, it is expected that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no dermal absorption data as well as no data available that suggests that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

Justification for classification or non-classification

Based on the available data for the assessment of repeated dose toxicity by oral, inhalation and dermal route and following CLP Regulation (EC) No 1272/2008, the substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2) is not classified.