m-toluidine

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Short description of key information:
In a OECD TG 422 Study male and female rats received 0,30, 100 and 300 mg/kg bw/day via gavage. The LOAEL (general toxicity is 30 mg/kg bw/day based on deposit pigmentation and extramedullary hematopoiesis in all examind animals. m-Toluidine induced implantation loss at 100 and 300 mg/kg bw/day . Any developmental toxicity including teratogenicity was not observed. The NOAELs for reproductive and developmental toxicity are considered to be 30 and 100 mg/kg bw/day, respectively (UNEP 2003)

Justification for selection of Effect on fertility via oral route:
This is the only reliable repeated dose toxicity study because it was performed according to the respective OECD Guideline under GLP condition and therefore evaluated with Klimisch score 1. Furthermore, the study meets the requirements of REACh for the tonnage band.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

initial bw (females): 200-247 g
initial bw (males): 267-361g
acclimisation: 5 days
temperature of the animal room: 24 °C
relative humidity: 55 %
lighting 12 hours light 12 hours dark
food ad libitum,
water ad libitum
pregnant females should be caged individually

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

Dosing of both sexes should begin 2 weeks prior to mating, continued through mating period.
Males: dosing continued up to the day when females are killed.
Females. dosing continued throughout pregnancy and up to day 4 of lactation.

Details on mating procedure:

One female to one male until pregnancy occurs; day 0 of pregnancy is defined as the day sperm is found.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Details not given.

Duration of treatment / exposure:

Males 42 days; females: from day 14 prior to mating to day 3 of lactation ( ca.42 days).

Frequency of treatment:

daily

Details on study schedule:

- Age at mating of the mated animals in the study: 10 weeks

Remarks:

Doses / Concentrations:
0, 30, 100, 300 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

13

Control animals:

yes, concurrent vehicle

Details on study design:

As requested by OECD TG 422.

Positive control:

No

Parental animals: Observations and examinations:

At least once per day:
--behavioual changes , signs of difficult or prolonged parturition, mortality and all signs of toxicity
--cage side observations: changes in skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system function
--food cinsumption, males and females should be weighed
--duration of gestation, examination of the litter as soon as possible: number and sex of pups, stillbirth, live birth, pup weight, and the presence of ggross anomalies
clinical examinations: hematology, clinical chemistry, urinalysis
pathology: gross necropsy, histopathology

Oestrous cyclicity (parental animals):

no data

Sperm parameters (parental animals):

no data

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: Number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities.

GROSS EXAMINATION OF DEAD PUPS:
yes, for external abnormalities

Postmortem examinations (parental animals):

Pathology: gross necropsy, histopathology

Postmortem examinations (offspring):

External malformation

Statistics:

yes, but methods not given

Reproductive indices:

number of mated pairs,
number of copulated pairs
copulation index,
number of pregnant animals,
fertility index,
pairing days undit copulation
implantation index
gestation index,
delivery index

Offspring viability indices:

number of pups born
number of pup alive
birth index
live birth index
sex ratio
number of pups alive on day 4
viability index
body weight of F1 pups up to day 4

Clinical signs:

effects observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Other effects:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

CLINICAL SIGNS AND MORTALITY
no deaths occurred during the study,
clinical signs included: brownish urine, increased salivation in males and females receiving 100 mg/kg bw/day and more
BODY WEIGHT AND WEIGHT GAIN
mean body weight gain during week 1 of the 300 mg/kg bw/day males and of the 100 and 300 mg/kg bw/day females was significantly lower than those of the controls
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
mean food consumption of the 300 mg/kg bw/day males and females was significantly reduced at week 1
HAEMATOLOGY (significant changes only)
no female data were mentioned
males
in the 100 and 300 mg/kg bw/day males values for erythrocyte count, hemoglobin concentration and hematocrit were decreased and mean corpuscular volume was increased.
At 300 mg/kg bw/day males mean corpuscular hemoglobin was significantly increased, mean corpuscular hemoglobin concentration was decreased.At 300 mgU/kg bw/day: in the differential blood picture segmented neutrophils and monocytes were significantly reduced as well as lymphocytes and platelet count
CLINICAL CHEMISTRY
no female data were mentioned
males
in the 100 and 300 mg/kg bw/day males the A/G ratio, , serum bilirubin, potassium and chloride levels significantly increased with the dose
in the 300 mg/kg bw/day males significant increases in serum glucose and total cholesterol levels
in the 300 mg/kg bw/day males significant decreases inalbumin, phospherous and GOT levek NEUROBEHAVIOUR
low motor activity and pale extremities in males and females given 300 mg/kg bw/day
ORGAN WEIGHTS
relative kidney weights of the 300 mg/kg bw7day males and females were significantly greater than those of the controls
PATHOLOGY
HISTOPATHOLOGY: NON-NEOPLASTIC
--from30 mg/kg bw/day onwards a slight increase in severity of pigmentation or extramedullary hematopoiesis in the spleen in males and females were noted
--Pigment deposites and extramedullary hematopoiesis in the liver and spleen and congestion in the spleen in males and females that received 100 mg/kg bw/day and more.
--Swelling of hepatocytes in centrilobular zone in 100 mg/kg bw/day males and in the 300 mg/kg bw/day males and females;
--deposits of pigment in the renal tubular epithelium in 300 mg /kg bw/day males and females
--eosiophilic droplets in the renal tubular epithelium in males given 100 mg/kg bw/day and more;
--regenerated renal tubules in females given 100 mg/kg bw/day and more
--focal hyperplasia of epithelium in the urinary bladder in the 300 mg/kg bw/day males
no effects on copulation index
no effect on fertility index
no effect on gestation index up to 100 mg/kg bw/day
gestation lenght no effect up to 100 mg/kg bw/day
total implantation loss at 300 mg/kg bw/day
implantation loss in 2/10 pregnant females at 100 mg/kg bw
no effect of delivery index up to 100 mg/kg bw/day

Dose descriptor:

NOAEL

Effect level:

30 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: based on implantation losses in all animals at 300 mg/kg bw/day and in 2 of 10 at 100 mg/kg bw/day

Dose descriptor:

LOAEL

Effect level:

30 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: Based on hemolytic anemia with deposit pigmentation and extramedulary hematopoiesis observed in all dosed rats

Remarks on result:

other: Generation: general toxicity

Clinical signs:

not specified

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:

not examined

VIABILITY (OFFSPRING)
up to and including 100 mg/kg bw/day : no effect

CLINICAL SIGNS (OFFSPRING)
no findings mentioned

BODY WEIGHT (OFFSPRING)
up to day 4 of lactation: no effect

GROSS PATHOLOGY (OFFSPRING)
up to and including of 100 mg/kg bw/day no findings

OTHER FINDINGS (OFFSPRING)
live birth index : no effects
viability index: no effect
sex ratio: no effects

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

100 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: no significant differences in the pup viability, pup weights and incidence of morphological abnormalities of pups

Reproductive effects observed:

not specified

Executive summary:

In the available OECD combined reated dose and reproductive /developmental toxicity screening study according to OECD TG 422 and GLP male and female rats received doses of 0, 30, 100 and 300 mg/kg bw/day 14 days before mating to 14 days after mating in males and for 14 days before mating to day 3 of lactation in females (MHLW 1995).

The LOAEL (general toxicity) is 30 mg/kg bw/day based on deposit pigmentation and extramedullary hematopoiesis in all examined animals (UNEP 2003). Reproductive organs were not affected by treatment.

No compound-related adverse effects were detected with regard to the mating performances at and dose levels. However, 2/10 pregnant females receiving 100 mg/kg bw/day and all eleven receiving 300 mg/kg bw/day showed total implant losses in utero. Therefore the NOAEL for reproductive toxicity is considered to be 30 mg/kg bw/day (UNEP 2003).

2/11 pregnant females receiving 30 mg/kg bw/day and 3/10 receiving 100 mg/kg bw/day did not show nursing activity obiously and 11 or more than half number of their pups died after birth, while all live offsprings of other dams in 30 and 100 mg/kg groups had normally developed up to 4 days. Therefore this death of pups is considered as a result of maternal toxicity, probably due to anemia. Furthermore, change of pup weights and incidence of morphologic abnormalities of pups were not significant in 30 and 100 mg/kg . The NOAEL for developmental toxicity is considered to be 100 mg/kg bw/day (UNEP 2003)

Effect on fertility: via oral route

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

30 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

This is the only reliable repeated dose toxicity study because it was performed according to the respective OECD Guideline under GLP condition and therefore evaluated with Klimisch score 1. Furthermore, the study meets the requirements of REACh for the tonnage band.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

In the available OECD combined repeated dose and reproductive /developmental toxicity screening study according to OECD TG 422 and GLP male and female rats received doses of 0, 30, 100 and 300 mg/kg bw/day 14 days before mating to 14 days after mating in males and for 14 days before mating to day 3 of lactation in females (MHLW 1995).

The LOAEL (general toxicity) is 30 mg/kg bw/day based on deposit pigmentation and extramedullary hematopoiesis in all examined animals (UNEP 2003).

Reproductive organs (testes, epididymides and ovar) are not affected by treatment.

No compound-related adverse effects were detected with regard to the mating performances at and dose levels. However, 2/10 pregnant females receiving 100 mg/kg bw/day and all eleven receiving 300 mg/kg bw/day showed total implant losses in utero. Therefore the NOAEL for reproductive toxicity is considered to be 30 mg/kg bw/day (UNEP 2003).

2/11 pregnant females receiving 30 mg/kg bw/day and 3/10 receiving 100 mg/kg bw/day did not show nursing activity obviously and 11 or more than half number of their pups diedafter birth, while all live offsprings of other dams in 30 and 100 mg/kg groups had normally developed up to 4 days. Therefore this death of pups is considered as a result of maternal toxicity , probably due to anemia. Furthermore, change of pup weights and incidence of morphologic abnormalities of pups were not significant in 30 and 100 mg/kg . The NOAEL for developmental toxicity is considered to be 100 mg/kg bw/day (UNEP 2003)

Effects on developmental toxicity

Description of key information

Short description of key information:

In a OECD TG 422 Study male and female rats received 0,30, 100 and 300 mg/kg bw/day via gavage. The LOAEL (general toxicity is 30 mg/kg bw/day based on deposit pigmentation and extramedullary hematopoiesis in all examined animals. m-toluidine induced implantation loss at 100 and 300 mg/kg bw/day . Any developmental toxicity including teratogenicity was not observed. The NOAELs for reproductive and developmental toxicity are considered to be 30 and 100 mg/kg bw/day, respectively (UNEP 2003).

 

Justification for selection of Effect on developmental toxicity: via oral route:
This is the only reliable repeated dose toxicity study bercause it was performed according to the respective OECD Guideline under GLP condition and therefore evaluated with Klimisch score 1. Furthermore, the study meets the requirements of REACh for the tonnage band.

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: OECD guideline 422 study and GLP

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: OECD TG 422

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Details on test animals or test system and environmental conditions:

Acclimatisation: 5 days
Temperature of the animal room: 22 °C
Relative humidity: 30-70 %
Lighting 12 hours light 12 hours dark
Food ad libitum,
Water ad libitum
Pregnant females should be caged individually

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

Dosing of both sexes should begin 2 weeks prior to mating , continued through mating period
Males: dosing continued up to the day when femalesare killed
Females. dosing continued throughout pregnancy and up to day 4 of lactation

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

method not mentioned

Details on mating procedure:

One female to one male until pregnancy occurs; day 0 of pregnancy is defined as the day sperm is found.

Duration of treatment / exposure:

Males 42 days;
Females: from day 14 prior to mating to day 3 of lactation ( ca.42 days).

Frequency of treatment:

daily

Duration of test:

Males 42 days; females: from day 14 prior to mating to day 3 of lactation ( ca.42 days).

No. of animals per sex per dose:

13

Control animals:

yes, concurrent vehicle

Details on study design:

As requested by OECD TG 422

Maternal examinations:

At least once per day:
--behavioual changes , signs of difficult or prolonged parturition, mortality and all signs of toxicity
--cage side observations: changes in skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system function
--food cinsumption, males and females should be weighed
--duration of gestation, examination of the litter as soon as possible: number and sex of pups, stillbirth, live birth, pup weight, and the presence of ggross anomalies
clinical examinations: hematology, clinical chemistry, urinalysis
pathology: gross necropsy, histopathology

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of resorptions: Yes

Fetal examinations:

- External examinations: Yes:

Statistics:

yes, but method not mentioned

Indices:

Copulation index
Fertility index
Gestation index
Implantation index
Delivery index
Birth index
Viability index

Historical control data:

No data

Details on maternal toxic effects:

Maternal toxic effects: yes

Details on maternal toxic effects:
no compound related effects adverse effects were detected with regared to the mating performance of any of the dosed rats. However, 2/10 pregnant females receiving 100 mg/kg bw/day and all the 11 females receiving 300 mg/kg bw/day showed toatal implantation losses in utero

Dose descriptor:

NOAEL

Effect level:

30 mg/kg bw/day (actual dose received)

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day (actual dose received)

Basis for effect level:

other: developmental toxicity

Dose descriptor:

LOAEL

Effect level:

30 mg/kg bw/day (actual dose received)

Basis for effect level:

other: other:

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects: no effects

Details on embryotoxic / teratogenic effects:
No significant differences in the pup viability, pup weights and incidence of morphological abnormalities of pups were apparent in the groups given 100 mg/kg bw/day or less when compared to controls.

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day (actual dose received)

Basis for effect level:

other: developmental toxicity

Dose descriptor:

LOAEL

Effect level:

30 mg/kg bw/day (actual dose received)

Basis for effect level:

other: other

Abnormalities:

not specified

Developmental effects observed:

not specified

Executive summary:

In the available OECD combined repeated dose and reproductive /developmental toxicity screening study according to OECD TG 422 and GLP male and female rats received doses of 0, 30, 100 and 300 mg/kg bw/day 14 days before mating to 14 days after mating in males and fron 14 days before mating to day 3 of lactation in females (MHLW 1995).

The LOAEL (general toxicity) is 30 mg/kg bw/day based on deposit pigmentation and extramedullary hematopoiesis in all examined animals (UNEP 2003).

No compound-related adverse effects were detected with regard to the mating performances at any dose level. However, 2/10 pregnant females receiving 100 mg/kg bw/day and all eleven receiving 300 mg/kg bw/day showed total implant losses in utero. Therefore the NOAEL for reproductive toxicity is considered to be 30 mg/kg bw/day (UNEP 2003).

2/11 pregnant females receiving 30 mg/kg bw/day and 3/10 receiving 100 mg/kg bw/day did not show nursing activity obviously and 11 or more than half number of their pups died after birth, while all live offsprings of other dams in 30 and 100 mg/kg groups had normally developed up to 4 days. Therefore this death of pups is considered as a result of maternal toxicity , probably due to anemia. Furthermore, change of pup weights and incidence of morphologic abnormalities of pups were not significant in 30 and 100 mg/kg . The NOAEL for developmental toxicity is considered to be 100 mg/kg bw/day (UNEP 2003).

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

100 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

This is the only reliable repeated dose toxicity study bercause it was performed according to the respective OECD Guideline under GLP condition and therefore evaluated with Klimisch score 1. Furthermore, the study meets the requirements of REACh for the tonnage band.

Additional information

In the available OECD combined repeated dose and reproductive /developmental toxicity screening study according to OECD TG 422 and GLP male and female rats received doses of 0, 30, 100 and 300 mg/kg bw/day 14 days before mating to 14 days after mating in males and for 14 days before mating to day 3 of lactation in females (MHLW 1995).

The LOAEL (general toxicity) is 30 mg/kg bw/day based on deposit pigmentation and extramedullary hematopoiesis in all examined animals (UNEP 2003).

No compound-related adverse effects were detected with regard to the mating performances at any dose level. However, 2/10 pregnant females receiving 100 mg/kg bw/day and all eleven receiving 300 mg/kg bw/day showed total implant losses in utero. Therefore the NOAEL for reproductive toxicity is considered to be 30 mg/kg bw/day (UNEP 2003).

2/11 pregnant females receiving 30 mg/kg bw/day and 3(10 receiving 100 mg/kg bw/day did not show nursing activity obviously and 11 or more than half number of their pups died after birth, while all live offsprings of other dams in 30 and 100 mg/kg groups had normally developed up to 4 days. Therefore this death of pups is considered as a result of maternal toxicity, probably due to anemia. Furthermore, change of pup weights and incidence of morphologic abnormalities of pups were not significant in 30 and 100 mg/kg . The NOAEL for developmental toxicity is considered to be 100 mg/kg bw/day (UNEP 2003)

Justification for selection of Effect on developmental toxicity: via oral route:
This is the only reliable repeated dose toxicity study bercause it was performed according to the respective OECD Guideline under GLP condition and therefore evaluated with Klimisch score 1. Furthermore, the study meets the requirements of REACh for the tonnage band.

Justification for classification or non-classification

Fertility assessment

Based on the available information fertility in females is only reduced in the presence of general toxicity mainly caused by hematotoxiciy and evidence of hemolytic anemia. Thus, the reduced fertility is most probably to be considered as a secondary effect.

Developmental assessment

Despite hematotoxicity in female and male rats all pups alive show normal development up to 4 days.

According to CLP classification criteria (Regulation (EC) No 1272/2008) a classification is not justified.