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

Developmental toxicity / teratogenicity

Administrative data

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

Data source

Reference

Reference Type:

publication

Title:

Studies on the Teratogenicity of Food Additives (4) Effects of Food Dye on the Preand Postnatal Development in Rats in Relation to Fetal Distribution

Author:

Shinsuke NAKAURA,

Year:

1975

Bibliographic source:

Food Hyg. Soc. 1975

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

242-355-6

EC Name:

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

Cas Number:

18472-87-2

Molecular formula:

C20H4Br4Cl4O5.2Na

IUPAC Name:

Sodium 2,3,4,5-tetrachloro-6-(2,4,5,7-tetrabromo-6-oxido-3-oxo-3H-xanthen-9-yl)benzoate

Test material form:

solid: particulate/powder

Details on test material:

IUPAC name: 3,4,5,6-Tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid
Molecular weight: 829.6388
Molecular formula: C20-H2-Br4-Cl4-O5.2Na
Smiles: [Na+].[Na+].[O-]c1c(Br)cc2c(Oc3c(Br)c([O-])c(Br)cc3C24OC(=O)c5c(Cl)c(Cl)c(Cl)c(Cl)c45)c1Br
Inchl: InChI=1S/C20H4Br4Cl4O5.2Na/c21-5-1-3-17(9(23)15(5)29)32-18-4(2-6(22)16(30)10(18)24)20(3)8-7(19(31)33-20)11(25)13(27)14(28)12(8)26;;/h1-2,29-30H;;/q;2*+1/p-2

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

Test animals

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

Administration / exposure

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

Doses / concentrations

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

Examinations

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

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

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

Maternal developmental toxicity

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.

Effect levels (maternal animals)

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.

Maternal abnormalities

Abnormalities:

no effects observed

Results (fetuses)

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.

Effect levels (fetuses)

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.

Fetal abnormalities

Abnormalities:

no effects observed

Overall developmental toxicity

Developmental effects observed:

no

Applicant's summary and conclusion

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.