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

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Effects on fertility

Description of key information

NOAEL was considered to be 920 mg/kg/day for F0 and F1 generation when Wistar female rats treated with 3,4,5,6-tetrachloro-2- (1,4,5,8-tetra bromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2).

Link to relevant study records

Reference

Endpoint:

one-generation reproductive toxicity

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:

The study was conducted to determine the reproductive toxicity potential of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen -9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.- 18472-87-2) when administered orally to wistar pregnant rats.

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): PHLOXINE B
- Molecular formula: C20H4Br4Cl4O5.2Na
- Molecular weight: 829.64 g/mol
- Substance type: Organic
- Physical state: Red-brown powder

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Nihon Rat Co. Ltd., Tokyo.
- Age at study initiation: 11 to 12 weeks
- Weight at study initiation:207.1-211.3g
- Fasting period before study: No data available
- Housing: Housed individually
- Diet (e.g. ad libitum): basal laboratory chow (NMF,Oriental Yeast Co., Tokyo) ad libitum
- Water (e.g. ad libitum): No data available
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25 ± 2 degC
- Humidity (%): 50-60 %
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available

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

Route of administration:

oral: feed

Vehicle:

other: basal laboratory chow

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Phloxine certified for food dye was obtained as powder from Hodogaya Chem. Co. Phloxine diet was prepared to contain 0, 0.3, 1.0 and 3.0% in a basal laboratory chow.

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Basal laboratory chow
- Concentration in vehicle: 0, 0.3, 1.0 and 3.0% (0, 280, 920, 2870 mg/kg/day)
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available

Details on mating procedure:

- M/F ratio per cage: No data available
- Length of cohabitation: Overnight
- Proof of pregnancy: Next morning those with vaginal plug or sperm in the vaginal smear were taken to be in day zero of pregnancy.
- After … days of unsuccessful pairing replacement of first male by another male with proven fertility. No data available
- Further matings after two unsuccessful attempts: No data available
- After successful mating each pregnant female was caged (how): No data available
- Any other deviations from standard protocol: No data available

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

19 days

Frequency of treatment:

Daily

Details on study schedule:

Food dye Red No.104(phloxin) was administered to Wistar pregnant rats at levels of 0, 0.3, 1, and 3% in diet during pregnancy and teratogenic effects and fetal and maternal organ distribution was examined. Suppression in maternal body weight gain and growth retardation in foetuses was observed in the highest dose group. No evidences of increase in fetal death and malformation were obtained in all groups. The postnatal development was maintained well without any adverse effects. However all newborn from 4 out of 5 dams in 3% group was killed by cannibalism within 2 days after birth.

Remarks:

0, 0.3, 1.0 and 3.0% (0, 280, 920, 2870 mg/kg/day)

No. of animals per sex per dose:

Total animals – females 80
0%(0 mg/kg/day): 20 female
0.3%(280 mg/kg/day): 20 female
1%(920 mg/kg/day): 20 female
3%(2870 mg/kg/day): 20 female

Control animals:

yes, concurrent vehicle

Parental animals: Observations and examinations:

Clinical sign, body weight and body weight gain, Food and Water Consumptions and food efficiency was examined.

Oestrous cyclicity (parental animals):

Any irregularity in estrous cyclicity was observed. Number of corpora lutea, inplantations and rate of nidations are also examined.

Sperm parameters (parental animals):

No data available

Litter observations:

Fetal mortality, litter size, body weight, body length, fetal resorption, sex ratio and tail length was observed. Motor activities, righting reflex, pinna reflex, pain response and startle response were also examined.

Postmortem examinations (parental animals):

Organ weight and gross pathology were observed.

Postmortem examinations (offspring):

Organ weight, visceral and skeletal anomalies, internal organ anomalies was examined.

Statistics:

No data available

Reproductive indices:

Gestation index, total implants index, birth index, weaning index and delivery index were examined

Offspring viability indices:

Viability in day 0, 4 and 8 week were examined.

Clinical signs:

no effects observed

Description (incidence and severity):

No toxic symptoms were observed in treated dams as compared to control.

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

At the highest dose level of 3%, a slight depression of weight gain were observed, but any other toxic symptoms were not observed throughout the period of pregnancy. No marked changes in 0.3 and 1% groups were noted.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

No effect was observed in food consumption of treated rats as compared to control.
The net amounts of phloxine ingested daily during pregnancy were 2.87g/kg for 3% group, 0.92 g/kg for 1% group and 0.28g/kg for 0.3% group, respectively.

Food efficiency:

effects observed, treatment-related

Description (incidence and severity):

At the highest dose level of 3%, decrease in food efficiency were observed.

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:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

No significant effect were observed in the number of corpora lutea or implantations and the rate of nidation of treated female rats as compared to control.

Key result

Dose descriptor:

NOAEL

Effect level:

920 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

clinical signs

body weight and weight gain

food consumption and compound intake

food efficiency

organ weights and organ / body weight ratios

gross pathology

histopathology: non-neoplastic

reproductive performance

other: No adverse effects was observed.

Critical effects observed:

no

Remarks on result:

not measured/tested

Critical effects observed:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

no mortality observed

Description (incidence and severity):

No evidences of increase in fetal mortality.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Significantly decrease in body weight was observed in female offspring at 2870 mg/kg/day dose groupe as compared to control.

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:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Significant effect were observed in lung weight of male rats at 920 mg/kgbw/day as compared to control. The observed changes were not remarkable.

Gross pathological findings:

no effects observed

Description (incidence and severity):

No gross pathological anomalies and skeleton were observed in treated offsprings as compared to control.

Histopathological findings:

effects observed, treatment-related

Description (incidence and severity):

No evidences of fetuses with external and internal malformations were obtained in all groups.
Prenatal development In 2870 mg/kg bw/day, slight increase in nucleus with deformed shapes and low stainability insternebrae and decreases in the number of metacarpus and coccygeal vertebrae were observed. These findings may suggest a retardation of ossification in fetal stage, but such changes were not observed in rats examined after weaning.

Other effects:

effects observed, treatment-related

Description (incidence and severity):

No significant difference between the phloxine-treated and control groups was found in the mean litter size, delivering rate or body weight of newborn at birth. In 3% group, however, all newborn from 4 out of 5 dams examined were killed by cannibalism of the dams within 2 days after birth. The mean litter sizes at the 4th and 8th weeks after birth were almost same among 0.3%, 1% and control groups. With the female of 3% group, significantly low body weight was obtained, but no appreciable failures.

Body lenght: Decrase in body lenght was observed in 2870 mg/kg/day treated offsprings as compared to control.

Tail lenght: No effect were observed on tail lenght of treated offspring as comparted to control.

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

Neurobehavioural parameters: No effect were observed on motor activities, righting reflex, pinna reflex, pain response and startle response of treated offsprings as compared to control.

Developmental immunotoxicity:

not specified

The postnatal development was maintained well without any adverse effects.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

920 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effect on survival, body weight, body lenght, tail lenght, organ weight, gross pathology and histopathology

Critical effects observed:

no

Remarks on result:

not measured/tested

Critical effects observed:

not specified

Reproductive effects observed:

no

Conclusions:

NOAEL was considered to be 920 mg/kg/day for F0 and F1 generation when Wistar female rats treated with 3,4,5,6-tetrachloro-2- (1,4,5,8-tetra bromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2).

Executive summary:

In a one generation reproductive toxicity study, wistar female rats treated with 3,4,5,6-tetrachloro-2- (1,4,5,8-tetra bromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2) in the concentrations of 0, 280, 920, 2870 mg/kg/day (0, 0.3, 1.0 and 3.0 %) during gestation orally by feed. No toxic symptoms were observed in treated rats. No effect on food consumption and water consumptions were observed in treated rats. Slight decrease in body weight gain and decrease in food efficiency and decrase in body lenght was observed in 2870 mg/kg/day treated offsprings as compared to control.No significant effects were observed in number of corpora lutea or implantations and the rate of nidation, organ weights and gross pathology in treated female rats as compared to control. No effect were observed on litter size and fetal mortality. Significantly decrease in body weight was observed in female offspring at 2870 mg/kg/day. Significant effect were observed in lung weight of male rats at 920 mg/kgbw/day as compared to control.The observed change in lung weight were not remarkalbe. No effect on tail lenght and gross pathology were observed in offspring. During prenatal development , slight increase in nucleus with deformed shapes and low stainability in sternebrae and decreases in the number of metacarpus and coccygeal vertebrae were observed. These findings may suggest a retardation of ossification in fetal stage, but such changes were not observed in rats examined after weaning.Therefore, NOAEL was considered to be 920 mg/kg/day for F0 and F1 generation when Wistar female rat treated with 3,4,5,6-tetrachloro-2- (1,4,5,8-tetra bromo-6-hydroxy-3- oxoxanthen-9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2) orally by feed.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

920 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The data is Klimicsh 2 and from peer-reviewed journal.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Various experimental studies from peer-reviewed journals has been investigated for reproductive toxicity to a greater or lesser extent for the test 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). The studies are summarized as below:

The one generation reproductive toxicity study was conducted by Shinsuke NAKAURA et al. (J. Food Hyg. Soc. Vol. 16, No. 1, pg 34-40, 1975). Wistar female rats treated with 3,4,5,6-tetrachloro-2- (1,4,5,8-tetra bromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2) in the concentrations of 0, 280, 920, 2870 mg/kg/day (0, 0.3, 1.0 and 3.0 %) during gestation orally by feed. No toxic symptoms were observed in treated rats. No effect on food consumption and water consumptions were observed in treated rats. Slight decrease in body weight gain and decrease in food efficiency and decrase in body lenght was observed in 2870 mg/kg/day treated offsprings as compared to control. No significant effects were observed in number of corpora lutea or implantations and the rate of nidation, organ weights and gross pathology in treated female rats as compared to control. No effect were observed on litter size and fetal mortality. Significantly decrease in body weight was observed in female offspring at 2870 mg/kg/day. Significant effect were observed in lung weight of male rats at 920 mg/kgbw/day as compared to control. The observed change in lung weight were not remarkalbe. No effect on tail lenght and gross pathology were observed in offspring. During prenatal development , slight increase in nucleus with deformed shapes and low stainability in sternebrae and decreases in the number of metacarpus and coccygeal vertebrae were observed. These findings may suggest a retardation of ossification in fetal stage, but such changes were not observed in rats examined after weaning. Therefore, NOAEL was considered to be 920 mg/kg/day for F0 and F1 generation when Wistar female rat treated with 3,4,5,6-tetrachloro-2- (1,4,5,8-tetra bromo-6-hydroxy-3- oxoxanthen-9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2) orally by feed.

 

Further, the study was published in a SCCNFP report (COLIPA n° C53, SCCNFP/0788/04, 23 April 2004), in which the reproductive toxicity potential of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy- 3-oxoxanthen-9-yl) benzoic acid (Phloxine) (CAS No.- 18472-87-2) was determined when administered orally to HanBrl: WIST(SPF) rats. Range finding study was conducted using 5 females (mated) HanBrl: WIST(SPF) rats. No toxic effects was observed on a range-finding study. 10, 50 and 250 mg/kg bw/day were selected as dose levels for a subsequent prenatal developmental toxicity study. The female HanBrl: WIST(SPF) rats were given Phloxine at concentrations of 0, 10, 50 and 250 mg/kg bw/day from day 6 through day 20 of gestation.

In the main study 1 low- and 1 mid-dose female died due to dosing error but no test-substance related effects were observed. Reddish faeces were observed in all test groups. Transient slight reductions inbody weight gainand food intake at 250 mg/kg bw/day were observed.No treatment related effects were observed on reproductive parameters tested. Reddish stomach/ intestinal content in mid- and high-dose group was observed.No treatment related effects were observed ongeneral foetal data, foetal visceral examination and foetal skeletal examination. 

The test substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy- 3-oxoxanthen-9-yl) benzoic acid (Phloxine) (CAS No.- 18472-87-2)elicited slight maternal toxicity at 250 mg/kg bw/day but was not embryotoxic or teratogenic at the doses tested. The NOAEL for maternal toxicity was considered to be 50 mg/kg bw/day whereas NOAEL for teratogenicity study was considered to be 250 mg/kg bw/day.

 

Moreover, the present study was conducted by M. SENO et al., (Fd Chem. Toxic. Vol. 22, No. 1 pp. 55-60, 1984) to determine the reproductive toxicity potential of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy- 3-oxoxanthen-9-yl) benzoic acid (Phloxine B) (CAS No.- 18472-87-2) when administered orally to pregnant Jcl:ICR mice. Pregnant Jcl:ICR mice were given Phloxine B in the diet at concentrations of 0, 1, 3 and 5%(0, 2000, 6000 and 10000 mg/kg/day)from the morning of day 6 through day 16 of gestation. The mice were killed on day 18 and foetuses were examined for external, visceral and skeletal anomalies. A significant decrease in body-weight gain was observed in all of the treated groups. Among the dams in the high-dose group, two maternal deaths, one abortion and a significant increase in liver weight were observed; none of these effects occurred at the lower dose levels. However, the 3% group, in which maternal toxicity was found only as a decrease in maternal bodyweight gain. A dose-related incidence of splitting of the cervical vertebral arches (nos 3-6) was noted in all of the treated groups, but this anomaly was not found in the controls. The total incidence of skeletal anomalies was also dose related and was significantly increased at the 3 and 5% dose levels. Therefore, NOAEL for maternal toxicity study (F0 -generation) was considered to be 2000.0 mg/kg/day (1%) and LOAEL for F1 generation was 2000.0 mg/kg bw/day (1%) when Jcl:ICR female mice were treated with 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) (CAS No. 18472-87-2). orally by feed from day 6 to 16 of gestation.

 

So, based on the above mentioned studies for target 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), it was found that no adverse effects on sexual function and fertility was observed. Therefore, according to CLP criteria, 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) cannot be classified as reproductive toxicant.

Effects on developmental toxicity

Description of key information

NOAEL was considered to be 920 mg/kg/day for F0 and F1 generation when Wistar female rats treated with 3,4,5,6-tetrachloro-2- (1,4,5,8-tetra bromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2).

Link to relevant study records

Reference

Endpoint:

developmental toxicity

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:

Effects of Food Dye Red No.104 (Phloxine) on the Pre and Postnatal Development in Rats in Relation to Fetal distribution were observed.

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): PHLOXINE B
- Molecular formula: C20H4Br4Cl4O5.2Na
- Molecular weight: 829.64 g/mol
- Substance type: Organic
- Physical state: Red-brown powder

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Nihon Rat Co. Ltd., Tokyo.
- Age at study initiation: 11 to 12 weeks
- Weight at study initiation:207.1-211.3g
- Fasting period before study: No data available
- Housing: Housed individually
- Diet (e.g. ad libitum): Basal laboratory chow (NMF,Oriental Yeast Co., Tokyo) ad libitum
- Water (e.g. ad libitum): No data available
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25 ± 2 degC
- Humidity (%): 50-60 %
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available

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

Route of administration:

oral: feed

Vehicle:

other: basal laboratory chow

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Phloxine certified for food dye was obtained as powder from Hodogaya Chem. Co. Phloxine diet was prepared to contain 0, 0.3, 1.0 and 3.0% in a basal laboratory chow.

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Basal laboratory chow
- Concentration in vehicle: 0, 0.3, 1.0 and 3.0% (0, 280, 920, 2870 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

Details on mating procedure:

- M/F ratio per cage: No data available
- Length of cohabitation: Overnight
- Proof of pregnancy: Next morning those with vaginal plug or sperm in the vaginal smear were taken to be in day zero of pregnancy.
- After … days of unsuccessful pairing replacement of first male by another male with proven fertility. No data available
- Further matings after two unsuccessful attempts: No data available
- After successful mating each pregnant female was caged (how): No data available
- Any other deviations from standard protocol: No data available

Duration of treatment / exposure:

19 days

Frequency of treatment:

Daily

Duration of test:

Upto day 57 after litter birth

Remarks:

0, 0.3, 1.0 and 3.0% (0, 280, 920, 2870 mg/kg/day)

No. of animals per sex per dose:

Total animals – females 80
0%(0 mg/kg/day): 20 female
0.3%(280 mg/kg/day): 20 female
1%(920 mg/kg/day): 20 female
3%(2870 mg/kg/day): 20 female

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Maternal examinations:

Clinical sign, body weight and body weight gain, Food and Water Consumptions and food efficiency was examined.

Ovaries and uterine content:

Number of corpora lutea, inplantations and rate of nidations are also examined.

Fetal examinations:

Fetal mortality, litter size, body weight, body length, fetal resorption , sex ratio and tail length, Motor activities, righting reflex, pinna reflex, pain response and startle response, Organ weight, visceral, skeletal anomalies and internal organ anomalies was examined.

Statistics:

No data available

Indices:

Gestation index, total implants index, birth index, weaning index and delivery index were examined.

Historical control data:

No data available

Clinical signs:

no effects observed

Description (incidence and severity):

No toxic symptoms were observed in treated dams as compared to control.

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

At the highest dose level of 3%, a slight depression of weight gain were observed, but any other toxic symptoms were not observed throughout the period of pregnancy. No marked changes in 0.3 and 1% groups were noted.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

No effect was observed in food consumption of treated rats as compared to control.
The net amounts of phloxine ingested daily during pregnancy were 2.87g/kg for 3% group, 0.92 g/kg for 1% group and 0.28g/kg for 0.3% group, respectively.

Food efficiency:

effects observed, treatment-related

Description (incidence and severity):

At the highest dose level of 3%, decrease in food efficiency were observed.

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:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

No significant differences between the phloxine-treated and control groups were found in the uterus and placental weight.

Gross pathological findings:

no effects observed

Description (incidence and severity):

No significant gross pathological changes were observed in treated female rats as compared to control.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Number of abortions:

not specified

Pre- and post-implantation loss:

not specified

Total litter losses by resorption:

not specified

Early or late resorptions:

not specified

Dead fetuses:

not specified

Changes in pregnancy duration:

not specified

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified

Changes in number of pregnant:

not specified

Other effects:

not specified

Details on maternal toxic effects:

Reproductive performance:No significant effect were observed in number of corpora lutea or implantations and the rate of nidation of treated female rats as compared to control.

Dose descriptor:

NOAEL

Effect level:

920 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

food efficiency

gross pathology

histopathology: non-neoplastic

organ weights and organ / body weight ratios

other: No adverse effect on reproductive performance was observed.

Abnormalities:

no effects observed

Fetal body weight changes:

not specified

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): effects observed, treatment-related

Reduction in number of live offspring:

not specified

Changes in sex ratio:

not specified

Changes in litter size and weights:

not specified

Changes in postnatal survival:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Visceral malformations:

no effects observed

Other effects:

not specified

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
Mortality: No effect were observed on litter size and fetal mortality of treated rats as compared to control.

Body weight:Significantly decrease in body weight was observed in female offspring at 2870 mg/kg/day dose group as compared to control.

Body lenght: Decrase in body lenght was observed in 2870 mg/kg/day treated offsprings as compared to control.

Tail lenght: No effect were observed on tail lenght of treated offspring as comparted to control.

Neurobehaveal parameters: No effect were observed on Motor activities, righting reflex, pinna reflex, pain response and startle response of treated offsprings as compared to control.

Organ weights:Significant effect were observed in lung weight of male rats at 920 mg/kgbw/day as compared to control. The observed change were not remarkalbe.

Gross pathology:No gross pathological anomalies were observed in treated offsprings as compared to control.

Histopathology:Prenatal development In 2870 mg/kg bw/day, slight increase in nucleus with deformed shapes and low stainability insternebrae and decreases in the number of metacarpus and coccygeal vertebrae were observed. These findings may suggest a retardation of ossification in fetal stage, but such changes were not observed in rats examined after weaning.

Postnatal development:No skeleton anomalies were observed in treated offsprings as compared to control.

Dose descriptor:

NOAEL

Effect level:

920 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effect on survival, body weight, body lenght, tail lenght, organ weight, gross pathology and histopathology.

Abnormalities:

no effects observed

Developmental effects observed:

no

Conclusions:

NOAEL was considered to be 920 mg/kg/day for F0 and F1 generation when Wistar female rats treated with 3,4,5,6-tetrachloro-2- (1,4,5,8-tetra bromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2).

Executive summary:

In a developmental toxicity study, wistar female rats treated with 3,4,5,6-tetrachloro-2 -(1,4,5,8-tetra bromo-6-hydroxy-3-oxoxanthen-9 -yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2) in the concentrations of 0, 280, 920, 2870 mg/kg/day (0, 0.3, 1.0 and 3.0 %) during gestation orally by feed. No toxic symptoms were observed in treated rats. No effect on food consumption and water consumptions were observed in treated rats. Slight decrease in body weight gain and decrease in food efficiency and decrase in body lenght was observed in 2870 mg/kg/day treated offsprings as compared to control. No significant effects were observed in number of corpora lutea or implantations and the rate of nidation, organ weights and gross pathology in treated female rats as compared to control. No effect were observed on litter size and fetal mortality. Significantly decrease in body weight was observed in female offspring at 2870 mg/kg/day. Significant effect were observed in lung weight of male rats at 920 mg/kgbw/day as compared to control.The observed change in lung weight were not remarkalbe. No effect on tail lenght and gross pathology were observed in offspring. During prenatal development , slight increase in nucleus with deformed shapes and low stainability in sternebrae and decreases in the number of metacarpus and coccygeal vertebrae were observed. These findings may suggest a retardation of ossification in fetal stage, but such changes were not observed in rats examined after weaning.

Therefore, NOAEL was considered to be 920 mg/kg/day for F0 and F1 generation when Wistar female rat treated with 3,4,5,6-tetrachloro-2- (1,4,5,8-tetra bromo-6-hydroxy-3 -oxoxanthen-9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2) orally by feed.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

920 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The data is Klimicsh 2 and from peer-reviewed journal.

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

The effect of oral 3,4,5,6-tetrachloro-2 -(1,4,5,8-tetra bromo-6-hydroxy-3-oxoxanthen-9 -yl)benzoic acid (CAS No.18472-87-2) exposure on mammalian development has been fully investigated. The studies are summarized as below:

In a developmental toxicity study conducted by Shinsuke NAKAURA et al. (J. Food Hyg. Soc. Vol. 16, No. 1, pg 34-40, 1975), wistar female rats treated with 3,4,5,6-tetrachloro-2 -(1,4,5,8-tetra bromo-6-hydroxy-3-oxoxanthen-9 -yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2) in the concentrations of 0, 280, 920, 2870 mg/kg/day (0, 0.3, 1.0 and 3.0 %) during gestation orally by feed. No toxic symptoms were observed in treated rats. No effect on food consumption and water consumptions were observed in treated rats. Slight decrease in body weight gain and decrease in food efficiency and decrase in body lenght was observed in 2870 mg/kg/day treated offsprings as compared to control. No significant effects were observed in number of corpora lutea or implantations and the rate of nidation, organ weights and gross pathology in treated female rats as compared to control. No effect were observed on litter size and fetal mortality. Significantly decrease in body weight was observed in female offspring at 2870 mg/kg/day. Significant effect were observed in lung weight of male rats at 920 mg/kgbw/day as compared to control.The observed change in lung weight were not remarkalbe. No effect on tail lenght and gross pathology were observed in offspring. During prenatal development , slight increase in nucleus with deformed shapes and low stainability in sternebrae and decreases in the number of metacarpus and coccygeal vertebrae were observed. These findings may suggest a retardation of ossification in fetal stage, but such changes were not observed in rats examined after weaning.

Therefore, NOAEL was considered to be 920 mg/kg/day for F0 and F1 generation when Wistar female rat treated with 3,4,5,6-tetrachloro-2- (1,4,5,8-tetra bromo-6-hydroxy-3 -oxoxanthen-9-yl)benzoic acid (Food dye Red No.104 / phloxine) (CAS No.18472-87-2) orally by feed. 

 

Moreover, the study published in a SCCNFP report (COLIPA n° C53, SCCNFP/0788/04, 23 April 2004). reveals that the developmental toxicity potential of 3,4,5,6-tetrachloro-2 -(1,4,5,8-tetrabromo-6-hydroxy- 3-oxoxanthen-9-yl) benzoic acid (Phloxine) (CAS No.- 18472-87-2) was determined when administered orally to HanBrl: WIST(SPF) rats. Range finding study was conducted using 5 females (mated) HanBrl: WIST(SPF) rats. No toxic effects was observed on a range-finding study. 10, 50 and 250 mg/kg bw/day were selected as dose levels for a subsequent prenatal developmental toxicity study. The female HanBrl: WIST(SPF) rats were given Phloxine at concentrations of 0, 10, 50 and 250 mg/kg bw/day from day 6 through day 20 of gestation.

In the main study 1 low- and 1 mid-dose female died due to dosing error but no test-substance related effects were observed. Reddish faeces were observed in all test groups. Transient slight reductions in body weight gain and food intake at 250 mg/kg bw/day were observed. No treatment related effects were observed on reproductive parameters tested. Reddish stomach/ intestinal content in mid- and high-dose group was observed.No treatment related effects were observed ongeneral foetal data, foetal visceral examination and foetal skeletal examination. The test substance 3,4,5,6-tetrachloro-2-(1,4,5,8- tetrabromo-6-hydroxy- 3-oxoxanthen-9-yl) benzoic acid (Phloxine) (CAS No.- 18472-87-2) elicited slight maternal toxicity at 250 mg/kg bw/day but was not embryotoxic or teratogenic at the doses tested. The NOAEL for maternal toxicity was considered to be 50 mg/kg bw/day whereas NOAEL for teratogenicity study was considered to be 250 mg/kg bw/day.

 

Thus, on the basis of above available studies and by comparing these with the CLP criteria, it can be concluded that no adverse effects on develop ment of the offspring was observed. Therefore the substance 3,4,5,6-tetrachloro-2 -(1,4,5,8-tetrabromo-6-hydroxy- 3-oxoxanthen-9-yl) benzoic acid (CAS No.- 18472-87-2) can be regarded as not owing a significant potential for developmental toxicity.

Justification for classification or non-classification

Based on the available data for the assessment of reproductive toxicity and following Regulation (EC) 1272/2008 no classification of 3,4,5,6- tetrachloro- 2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2) as reproductive toxicant is warranted.