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EC number: 207-586-9 | CAS number: 482-89-3
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
![](https://echa.europa.eu./o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Toxicity to reproduction
Administrative data
- Endpoint:
- three-generation reproductive toxicity
- Remarks:
- based on test type (migrated information)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1972/73
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions; no GLP
Data source
Referenceopen allclose all
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 973
- Report date:
- 1973
- Reference Type:
- publication
- Title:
- Toxicology of Indigo - a review
- Author:
- Ferber KH
- Year:
- 1 987
- Bibliographic source:
- J. Environ. Pathol. Toxicol. Oncol. 7, 73 -84 (1987)
- Reference Type:
- publication
- Title:
- Multi-generation Reproduction Studies with Certified Colors in Rats
- Author:
- Pierce E, Agersborg H, Borzelleca J, Burnett C, Eagle E, Ebert A, Kirschman J, Scala R
- Year:
- 1 974
- Bibliographic source:
- Toxicol Appl Pharmacol. 1974;29:121-122
Materials and methods
- Principles of method if other than guideline:
- Final version of Kirschman and Carpenter "A multi-generation study rats", dated January 7, 1972, amended on January 15 by Kirschman
- GLP compliance:
- no
- Limit test:
- no
Test material
- Reference substance name:
- 2-(1,3-dihydro-3-oxo-2H-indol-2-ylidene)-1,2-dihydro-3H-indol-3-one
- EC Number:
- 207-586-9
- EC Name:
- 2-(1,3-dihydro-3-oxo-2H-indol-2-ylidene)-1,2-dihydro-3H-indol-3-one
- Cas Number:
- 482-89-3
- Molecular formula:
- C16H10N2O2
- IUPAC Name:
- 2-(1,3-dihydro-3-oxo-2H-indol-2-ylidene)-1,2-dihydro-3H-indol-3-one
- Test material form:
- solid: particulate/powder
- Remarks:
- migrated information: powder
- Details on test material:
- - Name: D&C Blue #6
- Substance type: active ingredient
- Physical state: solid
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Wistar
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Harlan
Age at arrival: (P) 75 days
- Age at study initiation: (P) 11 - 12 weeks; (F1) 4 weeks
- Weight at study initiation: (P) Males: approx. mean 320 g; Females: approx. mean 238 g ; (F1) Males: approx. mean 164 g; Females: approx. mean 140 g
- Housing: before mating: 2/cage; during mating: 1 male:2 females; thereafter - females: single
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet: Purina Formulab TM Chow ad libitum
- Water: ad libitum
- Acclimation period: 8 days
Administration / exposure
- Route of administration:
- oral: feed
- Details on exposure:
- DIET PREPARATION
- Rate of preparation of diet (frequency): every 2 weeks
- Mixing appropriate amounts with (Type of food): Purina Formulab TM Chow
- Concentration in food: percentage color in diet = (dose level [mg/kg bw/day] x mean body weight [kg]) / (10 x mean food consumption [g/rat/day]) - Details on mating procedure:
- - M/F ratio per cage: 1:2
- Age at mating: (P) 1 - ca. 99 days; 2 - ca 169 days
(F1b) 1 - ca 100 days; 2 - ca 169 days; 3 - ca. 242 days
(F2b) 1 - ca. 102 days
- Treatment prior mating: 2 weeks
- Length of cohabitation: 15 days
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy - Analytical verification of doses or concentrations:
- not specified
- Duration of treatment / exposure:
- F0: 2-weeks prior mating until death - approx. 100 to 130 days
F1b/2b: from weaning until death - approx. 265 to 295/ 125 to 155 days - Frequency of treatment:
- continuously
- Details on study schedule:
- - F1 parental animals not mated until 10 weeks after selected from the F1 litters.
- Selection of parents from F1 generation when pups were 28 days of age.
- Age at mating of the mated animals in the study: (P) 1 - ca. 99 days; 2 - ca 169 days
(F1b) 1 - ca 100 days; 2 - ca 169 days; 3 - ca. 242 days
(F2b) 1 - ca. 102 days
Doses / concentrations
- Remarks:
- Doses / Concentrations:
5;50;150;500 mg/kg bw/day
Basis:
nominal in diet
- No. of animals per sex per dose:
- 10 males + 20 females
- Control animals:
- yes, concurrent no treatment
- Details on study design:
- 3 parallel control groups from different color testings were used as comparators
- Positive control:
- no
Examinations
- Parental animals: Observations and examinations:
- BEHAVIOUR AND APPEARENCE: Yes
- Time schedule: daily
BODY WEIGHT: Yes
- Time schedule for examinations: weekly
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/rat/day: Yes
- Compound intake calculated from the consumption and body weight gain data: Yes
-OTHER:
- Male and female fertility
- Gestation length
- F1b - third mating: half of dams per group were killed on Day 19: examination of
- uterus and contents examined: no of embryos, empty implantation sites, late and early resorption sites, any abnormal condition
- ovaries: no of corpora lutea
- necropsy in 5 rats/sex/group: (also true for F3a litter)
- marcoscopic examination and preservation of all tissues listed below for all dose groups
- adrenals, colon, heart, ileum, jejunum, kidney, liver, lung, ovary, uterus, spleen, stomach, testes, thyroid, urinary bladder, aorta, esophagus, eyes, mesenteric lymph node, pancreas, peripheral nerve, prostate, salivary gland, seminal vesicles, skeletal muscle, thymus, trachea
- microscopic examination of selected tissues in rats from the control and high dose groups: adrenals, colon, heart, ileum, jejunum, kidney, liver, lung, ovary, uterus, spleen, stomach, testes, thyroid, urinary bladder - Oestrous cyclicity (parental animals):
- no data
- Sperm parameters (parental animals):
- no data
- Litter observations:
- STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 10 pups/litter randomized; excess pups were killed and discarded.
PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring: number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities, other: necropsy see above
GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead. - Postmortem examinations (parental animals):
- see above: Observations and examinations
- Postmortem examinations (offspring):
- - F1b - third mating: half of dams per group were killed on Day 19: examination of
- uterus and contents examined: no of embryos, empty implantation sites, late and early resorption sites, any abnormal condition
- ovaries: no of corpora lutea
- necropsy in 5 rats/sex/group: (also true for F3a litter)
- marcoscopic examination and preservation of all tissues listed below for all dose groups
- adrenals, colon, heart, ileum, jejunum, kidney, liver, lung, ovary, uterus, spleen, stomach, testes, thyroid, urinary bladder, aorta, esophagus, eyes, mesenteric lymph node, pancreas, peripheral nerve, prostate, salivary gland, seminal vesicles, skeletal muscle, thymus, trachea
- microscopic examination of selected tissues in rats from the control and high dose groups: adrenals, colon, heart, ileum, jejunum, kidney, liver, lung, ovary, uterus, spleen, stomach, testes, thyroid, urinary bladder
SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at 21 days of age (except those of F2c generation delivered by cesarean section on Day 19 of pregnancy).
- These animals were subjected to postmortem examinations macroscopic and/or microscopic examination as follows:
NECROPSY
- Gross necropsy consisted of external and internal examinations
- F3a litter: necropsy in 5 rats/sex/group:
- marcoscopic examination and preservation of all tissues listed below for all dose groups
- adrenals, colon, heart, ileum, jejunum, kidney, liver, lung, ovary, uterus, spleen, stomach, testes, thyroid, urinary bladder, aorta, esophagus, eyes, mesenteric lymph node, pancreas, peripheral nerve, prostate, salivary gland, seminal vesicles, skeletal muscle, thymus, trachea
- microscopic examination of selected tissues in rats from the control and high dose groups: adrenals, colon, heart, ileum, jejunum, kidney, liver, lung, ovary, uterus, spleen, stomach, testes, thyroid, urinary bladder - Statistics:
- Quantitative variables of 3 control and 4 dosage groups: intercomparison with Bartlett's homogeneity of variances, analysis of variance, Duncan's multiple range tests
In case of inhomogeneity: F-test --> if not significant: Student's t-test
--> if significant: Cochran t-test or sum of ranks
Levels of significance: a - 0.05 > P > 0.01
b - 0.01 > P > 0.001
c - P < 0.001
Statistical methods used are described in "Selection of the valid number of sampling units and a consideration of their combination in toxicological studies involving reproduction, teratogenesis or carcinogenesis! Weil CS. Food and Cosmetics Toxicology 8:177-182; 1970 - Reproductive indices:
- Fertility index
Gestation index
Gestation survival index - Offspring viability indices:
- 4-day survival index
14-day survival index
21-day survival index
Lactation index
Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
- Clinical signs:
- no effects observed
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Organ weight findings including organ / body weight ratios:
- not examined
- Histopathological findings: non-neoplastic:
- no effects observed
- Other effects:
- no effects observed
Reproductive function / performance (P0)
- Reproductive function: oestrous cycle:
- not specified
- Reproductive function: sperm measures:
- not examined
- Reproductive performance:
- no effects observed
Details on results (P0)
Effect levels (P0)
open allclose all
- Dose descriptor:
- NOEL
- Effect level:
- 500 mg/kg bw/day
- Sex:
- male/female
- Basis for effect level:
- other: see 'Remark'
- Dose descriptor:
- NOEL
- Effect level:
- 500 mg/kg bw/day
- Sex:
- male/female
- Basis for effect level:
- other: clinical signs; mortality; body weight; gross pathology; histopathology
- Remarks on result:
- other: Generation: F3 (migrated information)
Results: F1 generation
General toxicity (F1)
- Clinical signs:
- no effects observed
- Mortality / viability:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Sexual maturation:
- no effects observed
- Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- no effects observed
- Histopathological findings:
- no effects observed
Details on results (F1)
Effect levels (F1)
- Dose descriptor:
- NOEL
- Generation:
- F1
- Effect level:
- 500 mg/kg bw/day
- Sex:
- male/female
- Basis for effect level:
- other: see 'Remark'
Results: F2 generation
Effect levels (F2)
- Dose descriptor:
- NOEL
- Generation:
- F2
- Effect level:
- 500 mg/kg bw/day
- Sex:
- male/female
- Basis for effect level:
- other: see 'Remark'
Overall reproductive toxicity
- Reproductive effects observed:
- not specified
Applicant's summary and conclusion
- Conclusions:
- No test item-related effects were observed, only random, not deleteriuous effects were associated with the inclusion of indigo in the diet of rats for 3 generations.
Indigo does neither have adverse effects on the reproductive system and fertility of parental rats and theit offspring, nor on the development of the offspring. - Executive summary:
10 males and 20 females of each generation were dosed with 0, 0, 0, 5, 50, 150, 500 mg/kg bw/day of D&C Blue #6 and mated. The parent generation (F0) rats were bred twice, the F1b were bred thrice, and the F2b were bred once. The criteria of effect included the following indices: fertility, gestation, gestation survival, 4 -/14 -/21 -day survival. Furthermore, the diet consumption and body weights of the parent rats and their progeny data were examined as were the number of resorption sites and corpora lutea at the 19-day kill of the third mating of half of the F1b dams.
The differences between dosed and control groups were neither dose-related, nor progressive, nor were they indicative of mean or median values exceeding those of the controls. It is therefore concluded that only random, not deleteriuous effects were associated with the inclusion of indigo in the diet of rats for 3 generations.
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