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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
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Endpoint summary
Administrative data
Key value for chemical safety assessment
Effects on fertility
Link to relevant study records
- 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
- 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
- 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 - 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 - 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
- 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 - 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: oestrous cycle:
- not specified
- Reproductive function: sperm measures:
- not examined
- Reproductive performance:
- no effects observed
- 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)
- 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
- Dose descriptor:
- NOEL
- Generation:
- F1
- Effect level:
- 500 mg/kg bw/day
- Sex:
- male/female
- Basis for effect level:
- other: see 'Remark'
- Dose descriptor:
- NOEL
- Generation:
- F2
- Effect level:
- 500 mg/kg bw/day
- Sex:
- male/female
- Basis for effect level:
- other: see 'Remark'
- Reproductive effects observed:
- not specified
- 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.
Reference
Effect on fertility: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 500 mg/kg bw/day
- Study duration:
- chronic
- Species:
- rat
Effect on fertility: via inhalation route
- Endpoint conclusion:
- no study available
Effect on fertility: via dermal route
- Endpoint conclusion:
- no study available
Additional information
Three Generation Study With Indigo In Rats:
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 deleterious effects were
associated with the inclusion of indigo in the diet of rats for
3 generations.
Short description of key information:
No effects on reproduction process observed
Effects on developmental toxicity
Description of key information
No effects on intra-uterine development observed
Link to relevant study records
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1971/1972
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Acceptable, well-documented study report which meets basic scientific principles
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Deviations:
- yes
- Remarks:
- no maternal toxicity observed
- GLP compliance:
- not specified
- Limit test:
- no
- Species:
- rat
- Strain:
- Sprague-Dawley
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River (obtained as weanlings)
- Age at study initiation: satisfactory age for mating (about 8 weeks)
- Weight at study initiation: -
- Fasting period before study: -
- Housing: single
- Diet: Purina Laboratory Chow ad libitum
- Water: tap ad libitum
- Acclimation period: ca. 5 weeks - Route of administration:
- oral: gavage
- Vehicle:
- other: methylcellulose 0.5%
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
suspension in 0.5% methylcellulose
VEHICLE
- Dow Methocel 90 HG, 15000 CPS
- Justification for use and choice of vehicle (if other than water): TS is insoluble in water
- Concentration in vehicle: 50 mg/mL
- Amount of vehicle: 10 mL/kg (control and high dose), 3.2 mL/kg, 1 mL/kg
- Lot/batch no.: MM 101012K
POSITIVE CONTROL
- Acetylsalicylic acid (Aspirin)
- Supplier: Mallinckrodt - Analytical verification of doses or concentrations:
- not specified
- Details on mating procedure:
- - Impregnation procedure: cohoused
- 1 M/1 F per cage
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy - Duration of treatment / exposure:
- Day 6 through 15 of gestation
- Frequency of treatment:
- daily
- Duration of test:
- 21 days
- Remarks:
- Doses / Concentrations:
50, 160, 500 mg/kg/day
Basis:
other: nominal in vehicle - No. of animals per sex per dose:
- 20 pregnant females
- Control animals:
- yes, concurrent vehicle
- other: positive control: aspirin
- Details on study design:
- 3 dosage levels were run concurrently with 2 negative (vehicle) and one positive (Aspirin) control groups
- Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: -
BODY WEIGHT: Yes
- Time schedule for examinations: Days 0, 6, 15, 20 of gestation
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: uterus - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: No data
- Number of implantations: Yes
- Number of resorptions: Yes
- Number viable and dead fetuses: Yes
- Distribution by sex: Yes
- Mean litter size: Yes - Fetal examinations:
- - Mean fetal weight
- Determination of sex
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: one-third per litter
- Skeletal examinations: Yes: two-third per litter - Statistics:
- Frequency of occurrence and seriousness of anomaly compared to control groups
- Indices:
- no data
- Historical control data:
- no data
- Details on maternal toxic effects:
- Maternal toxic effects:no effects
Details on maternal toxic effects:
- - Dose descriptor:
- NOEL
- Effect level:
- 500 mg/kg bw/day
- Based on:
- act. ingr.
- Basis for effect level:
- other: developmental toxicity
- Dose descriptor:
- NOEL
- Effect level:
- 500 mg/kg bw/day (nominal)
- Based on:
- act. ingr.
- Basis for effect level:
- other: maternal toxicity
- Details on embryotoxic / teratogenic effects:
- Embryotoxic / teratogenic effects:no effects
Details on embryotoxic / teratogenic effects:
- - Dose descriptor:
- NOEL
- Effect level:
- 500 mg/kg bw/day (nominal)
- Based on:
- act. ingr.
- Basis for effect level:
- other: teratogenicity
- Abnormalities:
- not specified
- Developmental effects observed:
- not specified
- Conclusions:
- D&C Blue No. 6 administered by gavage to groups of 20 female rats each during days 6 through 15 of gestation at dose levels of 50, 160, and 500 mg/kg/day was without effect on reproductive performance, maternal weight gain, and fetal development.
- Executive summary:
Groups of 20 pregnant female Charles River CD rats received suspensions of D&C Blue No. 6 by stomach tube at doses of 500, 160 and 50 mg/kg/day from day 6 through day 15 of gestation. Comparable groups received either the methylcellulose suspending vehicle as negative controls or aspirin, 250 mg/kg/day as positive controls. On day 20 of gestation Caesarean section was performed, the uterus examined and fetuses processed for subsequent examination for skeletal or visceral anomalies.
On the basis of numbers of viable and dead fetuses, resorption sites, mean fetal weight, distribution by sex, mean litter size, frequency of skeletal and visceral or structural anomalies; weight gain of pregnant females: Indigo was without effect on reproductive performance, maternal weight gain and fetal development.
Reference
Effect on developmental toxicity: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 500 mg/kg bw/day
- Study duration:
- subacute
- Species:
- rat
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
Teratogenicity Indigo Rat:
Groups of 20 pregnant female
Charles River CD rats received suspensions of D&C Blue No. 6 by stomach
tube at doses of 500, 160 and 50 mg/kg/day from day 6 through day 15 of
gestation. Comparable groups received either the methylcellulose
suspending vehicle as negative controls or aspirin, 250 mg/kg/day as
positive controls. On day 20 of gestation Caesarean section was
performed, the uterus examined and fetuses processed for subsequent
examination for skeletal or visceral anomalies.
On the basis of numbers of viable and dead fetuses, resorption sites,
mean fetal weight, distribution by sex, mean litter size, frequency of
skeletal and visceral or structural anomalies; weight gain of pregnant
females: Indigo was without effect on reproductive performance, maternal
weight gain and fetal development.
Teratogenicity Indigo Rabbit:
Groups of 10 pregnant female New
Zealand white rabbits received suspensions of D&C Blue No. 6 by stomach
tube at doses of 500, 160 and 50 mg/kg/day from day 6 through day 18 of
gestation. Comparable groups received either the methylcellulose
suspending vehicle as negative controls or thalidomide at 100 mg/kg/day
as positive controls. On day 29 of gestation Caesarean section was
performed, the uterus examined and fetuses autopsied for detection of
visceral anomalies. Subsequently, the eviscerated carcasses were
processed for examination of skeletal anomalies.
On the basis of numbers of viable and dead fetuses, resorption sites,
mean fetal weight, distribution by sex, mean litter size, frequency of
skeletal and visceral or structural anomalies; weight gain of pregnant
females: Indigo was without effect on reproductive performance, maternal
weight gain and fetal development.
Justification for classification or non-classification
No effects on reproduction observed, no classification necessary
Additional information
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
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