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Key value for chemical safety assessment

Effects on fertility

Description of key information
no data
Link to relevant study records
Reference
Endpoint:
toxicity to reproduction
Remarks:
other: 28 d repeated dose study
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Principles of method if other than guideline:
Reinblau RLW Tr. was administered by oral gavage to male and female SD rats for 28 days at dose levels of 0, 8, 40, 200, and 1000 mg/kg to asses its toxicity and reversibility. At the end of the treatment period or an additional revovery period all animals were killed at day 29 or day 43 and necropsied; gross and microscopic examination of all major organs, including reproductive organs.
GLP compliance:
yes
Species:
rat
Strain:
other: Crj:CD (SD) IGS rats (48 males and 48 females) were obtained from Charles Liver Laboratories Japan Ltd.
Sex:
male/female
Route of administration:
oral: gavage
Vehicle:
other: The test substance was suspended in the vehicle (0.5% CMC-Na solution)
Details on mating procedure:
not applicable
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
28 days
Frequency of treatment:
once daily in the morning for 28 days.
Details on study schedule:
6 male and 6 female rats of the control, 8, 40, 200 and 1000 mg/kg bw dose group were necropsied on day 29; additionally 6 male and 6 female rats of the control, 200, and 1000 mg/kg bw dose group were necropsied after the recovery period on day 43.
Remarks:
Doses / Concentrations:
0, 8, 40, 200, or 1000 mg/kg
Basis:
other: nominal value
No. of animals per sex per dose:
control: 6 male and 6 female rats in the control, 8, 40, 200 and 1000 mg/kg bw dose group
recovery group: 6 male and 6 female rats in the control, 200, and 1000 mg/kg bw dose group
Control animals:
yes, concurrent vehicle
Reproductive effects observed:
not specified

There were no test substance-related toxic changes in organs weights of testis and ovaries. No histopathological changes were found in testis, ovaries and all other examined organs. Additional no changes were seen relating to in clinical signs, body weight, food consumption, hematology, blood chemistry and urinalysis.

Conclusions:
Fertility is not affected at doses of 1000 mg/kg bw for 28 days
Executive summary:

Reinblau RLW Tr. was administered by oral gavage to male and female SD rats for 28 days at dose levels of 0, 8, 40, 200, and 1000 mg/kg to assess its toxicity and reversibility. At the end of the treatment period or an additional revovery period all animals were killed at day 29 or day 43 and necropsied; gross and microscopic examination of: all major organs, including reproductive organs.

There were no test substance-related toxic changes in clinical sign, body weight, food consumption, hematology, blood chemistry, urinalysis, organ weight, necropsy, or histopathology.

No evidence of toxicity to reproduction was observed in a subacute repeated dose study as no treatment-related changes were observed for any reproductive organ investigated during macroscopic and microscopic examination. On the basis of this study no effects on fertility were expected (NOEL, rat: 1000 mg/kg bw/day).

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
Study duration:
subacute
Species:
rat
Quality of whole database:
GLP guideline study
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

No evidence of toxicity to reproductive organs was observed in a subacute repeated dose study as no treatment-related changes were observed for any reproductive organ investigated during macroscopic and microscopic examination. On the basis of this study no effects on fertility were expected (NOEL, rat: 1000 mg/kg bw/day).


Short description of key information:
There was no fertility study with 1,4-bis[(2-ethyl-6-methylphenyl)amino]anthraquinone available. No effects on reproductive organs were observed in a 28 day study in rats. The pathologic evaluation consisted of organ weight, gross and microscopic examination of reproductive organs, incl. tetes and ovaries. No treatment-related changes were observed for any reproductive organ investigated during macroscopic and microscopic examination of all major organs (NOAEL, rat: 1000 mg/kg bw/day; males and females).

Justification for selection of Effect on fertility via oral route:
Key study is used (only available study)

Effects on developmental toxicity

Description of key information
A study for effects on embryo-fetal development according to OECD TG 414 and GLP was conducted in the Sprague-Dawley rat by oral gavage administration of 1000 mg Macrolex Blau 3R/kg/day (CAS no. 41611-76-1). 
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Principles of method if other than guideline:
One group of 22 females received Macrolex Blau 3R at the limit dose of 1000 mg/kg/day by daily oral gavage administration at a dose volume of 10 mL/kg/day from Day 6 to 19 after mating. A similarly constituted control group received the vehicle, 0.5% CMC-Na solution, according to the same dose volume and regimen. Animals were killed on Day 20 after mating for reproductive assessment and fetal examination.
Clinical observations, body weight and food consumption were recorded. Adult females were examined macroscopically at necropsy on Day 20 after mating with the weight of the gravid uterus recorded. All fetuses were weighed, subjected to an external macroscopic examination and then subsequently to detailed internal visceral or skeletal examination.
GLP compliance:
yes
Limit test:
yes
Species:
rat
Strain:
other: Crl:CD(SD)
Route of administration:
oral: gavage
Vehicle:
other: 0.5% CMC-Na solution
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
Mating
Male/female ratio: 1:1 with identified stock males.
Daily checks for evidence of mating: Ejected copulation plugs in cage tray and vaginal smears were checked for the presence of sperm.
Day 0 of gestation: When positive evidence of mating was detected.
A colony of stud males was maintained specifically for the purpose of mating; these animals were not part of the study and were maintained as stock animals.
Duration of treatment / exposure:
Females were treated from Day 6 to Day 19 (inclusive) after mating, once daily at approximately the same time each day.
Frequency of treatment:
Once daily
Duration of test:
21 days
Remarks:
Doses / Concentrations:
1000 mg/kg/day
Basis:
nominal conc.
No. of animals per sex per dose:
1000 mg/kg/day: 22 females
0 mg/kg/day: 22 females (control group)
Control animals:
yes, concurrent vehicle
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects
Abnormalities:
not specified
Developmental effects observed:
not specified

Treatment of pregnant female Sprague Dawley rats with Macrolex Blau 3R daily from Day 6 to Day 19 after mating, inclusive, at the limit dose of 1000 mg/kg/day was well tolerated and elicited no deaths and no toxicity related changes in clinical condition of the adult females.

At 1000 mg/kg/day a total of 17 females were recorded with the clinical sign of blue staining of the tail. At necropsy several females receiving 1000 mg/kg/day showed blue colouration of gastro-intestinal contents, three females showed blue colouration of rectal tissue, 5 females were recorded with blue coloration of stomach tissue and 17 females were confirmed as having blue colouration of the tail. The blue staining observed was considered discolouration caused by the test material itself (a blue dye) and is considered not to represent toxicity.

Body weight gain and food consumption during gestation were unaffected by treatment and there were no other maternal findings observed at macroscopic examination that were considered to relate to treatment.

There was no effect of treatment at 1000 mg/kg/day upon embryo-fetal survival or placental weights, and fetal development was also unaffected by treatment at this limit dose, with the incidence of major and minor abnormalities and skeletal variants showing no relationship to treatment.

Mean male, female and overall fetal weights at 1000 mg/kg/day were lower than Controls, but the difference was considered marginal (approx. 5-6%) and was clearly not adverse since embryo fetal survival and development were unaffected by treatment.

Based on the results of this embryo-fetal developmental toxicity study in rats, it was concluded that the No-Observed-Effect-Level (NOEL) of Macrolex Blau 3R for maternal toxicity was the limit dose of 1000 mg/kg/day.

For embryo-fetal development, the limit dose of 1000 mg/kg/day was considered to be the No-Observed-Adverse-Effect-Level (NOAEL), when administered during the organogenesis and fetal growth phases of gestation in the rat, as there was a marginal reduction in fetal weight which was considered not adverse due to there being no effect of treatment upon embryo-fetal survival or development.

Conclusions:
Based on the results of this embryo-fetal developmental toxicity study in rats, it was concluded that the No-Observed-Effect-Level (NOEL) of Macrolex Blau 3R for maternal toxicity was the limit dose of 1000 mg/kg/day.
For embryo-fetal development, the limit dose of 1000 mg/kg/day was considered to be the No-Observed-Adverse-Effect-Level (NOAEL), when administered during the organogenesis and fetal growth phases of gestation in the rat, as there was a marginal reduction in fetal weight which was considered not adverse due to there being no effect of treatment upon embryo-fetal survival or development.
Executive summary:

The objective of this study was to assess the influence of Macrolex Blau 3R, a blue powder, on embryo-fetal survival and development when administered during the organogenesis and fetal growth phases of pregnancy in the Sprague Dawley CD rat.

One group of 22 females received Macrolex Blau 3R at the limit dose of 1000 mg/kg/day by daily oral gavage administration at a dose volume of 10 mL/kg/day from Day 6 to 19 after mating. A similarly constituted Control group received the vehicle, 0.5% CMC-Na solution, according to the same dose volume and regimen. Animals were killed on Day 20 after mating for reproductive assessment and fetal examination.

Clinical observations, body weight and food consumption were recorded. Adult females were examined macroscopically at necropsy on Day 20 after mating with the weight of the gravid uterus recorded. All fetuses were weighed, subjected to an external macroscopic examination and then subsequently to detailed internal visceral or skeletal examination.

Treatment of pregnant female Sprague Dawley rats with Macrolex Blau 3R daily from Day 6 to Day 19 after mating, inclusive, at the limit dose of 1000 mg/kg/day was well tolerated and elicited no deaths and no toxicity related changes in clinical condition of the adult females.

At 1000 mg/kg/day a total of 17 females were recorded with the clinical sign of blue staining of the tail. At necropsy several females receiving 1000 mg/kg/day showed blue colouration of gastro-intestinal contents, three females showed blue colouration of rectal tissue, 5 females were recorded with blue coloration of stomach tissue and 17 females were confirmed as having blue colouration of the tail. The blue staining observed was considered discolouration caused by the test material itself (a blue dye) and is considered not to represent toxicity.

Body weight gain and food consumption during gestation were unaffected by treatment and there were no other maternal findings observed at macroscopic examination that were considered to relate to treatment.

There was no effect of treatment at 1000 mg/kg/day upon embryo-fetal survival or placental weights, and fetal development was also unaffected by treatment at this limit dose, with the incidence of major and minor abnormalities and skeletal variants showing no relationship to treatment.

Mean male, female and overall fetal weights at 1000 mg/kg/day were lower than Controls, but the difference was considered marginal (approx. 5-6%) and was clearly not adverse since embryo fetal survival and development were unaffected by treatment.

Based on the results of this embryo-fetal developmental toxicity study in rats, it was concluded that the No-Observed-Adverse- Effect-Level (NOAEL) of Macrolex Blau 3R for maternal toxicity and embryo-fetal development was the limit dose of 1000 mg/kg/day.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
Study duration:
subacute
Species:
rat
Quality of whole database:
GLP guideline study
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

A developmental toxicity study according to OECD TG 414 was conducted in rats. Test stubstance was administered during the organogenesis and fetal growth phases of pregnancy in the Sprague Dawley CD rat. One group of 22 females received Macrolex Blau 3R at the limit dose of 1000 mg/kg/day by daily oral gavage administration at a dose volume of 10 mL/kg/day from Day 6 to 19 after mating. A similarly constituted Control group received the vehicle alone. Animals were killed on Day 20 after mating for reproductive assessment and fetal examination. Clinical observations, body weight and food consumption were recorded. Adult females were examined macroscopically at necropsy on Day 20 after mating with the weight of the gravid uterus recorded. All fetuses were weighed, subjected to an external macroscopic examination and then subsequently to detailed internal visceral or skeletal examination.

Based on the results of this embryo-fetal developmental toxicity study in rats, it was concluded that the No-Observed-Adverse- Effect-Level (NOAEL) of Macrolex Blau 3R for maternal toxicity and embryo-fetal development was the limit dose of 1000 mg/kg/day.


Justification for selection of Effect on developmental toxicity: via oral route:
Key study is used (only available study)

Toxicity to reproduction: other studies

Additional information

no data

Justification for classification or non-classification

Based on the available study for repeated dose toxicity a non-classification for fertility is justified. Additional a developmental toxicity study according OECD TG 414 is available. The NOAEL in this study was 1000 mg/kg/bw of the test substance for maternal and developmental toxicity.

Overall, a classification for toxicity on reproduction is not justified for Macrolex Blau 3R (CAS no. 41611-76-1).