3-methyl-1-vinyl-1H-imidazolium chloride

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2012-02-09 and 2013-05-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-conform study in accordance to OECD guideline and US EPA guideline. The test substance analogue was used. For read-across justification, please refer to section 13.

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3650 (Combined Repeated Dose Toxicity Study With the Reproduction/Developmental Toxicity Screening Test) 2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Age at study initiation: 12-13 weeks
- Weight at study initiation: F0 generation: male animals: 339.0 g - 372.4 g, female animals: 191.1 g - 225.1 g
- Housing:
Individually in Makrolon type M III cages, with the following exceptions: a) During overnight matings, male and female mating partners were housed together in Makrolon type M III cages. b) Pregnant animals and their litters were housed together until PND 4.
- Diet: ad libitum, Kliba maintenance diet mouse/rat “GLP” meal (Provimi Kliba SA, Kaiseraugst, Switzerland)
- Water: ad libitum
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: For the preparation of the administration solutions the test substance was weighed in a calibrated beaker depending on the dose group, topped up with drinking water and subsequently intensely mixed with a magnetic stirrer until it was completely dissolved. The analytical results indicated that the test substance is stable in drinking water over a period of 7 days.

DIET PREPARATION
- Rate of preparation of diet (frequency): at the beginning of the administration period and thereafter in intervals

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: overnight for a maximum of two weeks
- Proof of pregnancy: vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: individually
- Any other deviations from standard protocol: none

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The sample preparation were analytically verified by using HPLC method coupled with a DAD. Therefore, approx. 0.05 g of the test substance was dissolved to 100 mL usind drinking water. All determined concentrations were in the range between 90% and 110% of the nominal concentration, demonstrationg the correctness of the preparations.

Duration of treatment / exposure:

females: 49 days
males: 34 days

Frequency of treatment:

once daily

Remarks:

Doses / Concentrations:
0, 100, 300, 1000 mg/kg bw/d
Basis:
nominal in water

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Positive control:

not applicable

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once daily
- Cage side observations checked: any signs of morbidity, pertinent behavioral changes and signs of overt toxicity
- The parturition and lactation behavior of the dams was generally evaluated in the mornings in combination with the daily clinical inspection of the dams. Only particular findings (e.g. inability to deliver) were documented on an individual dam basis.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once prior to the first administration and at weekly intervals during the administration period

BODY WEIGHT: Yes
- Time schedule for examinations: once a week
- The following exceptions are notable for the female animals:
a) During the mating period the parental females were weighed on the day of positive evidence of sperm (gestation day 0) and on gestation days 7, 14 and 20.
b) Females with litter were weighed on the day of parturition (postnatal day 0) and on postnatal day 4.

FOOD CONSUMPTION: Yes
- once a week
- Exceptions:
a) Food consumption was not determined after the 2nd premating week (male parental animals) and during the mating period (male and female F0 animals).
b) Food consumption of the F0 females with evidence of sperm was determined on gestation days (GD) 0, 7, 14 and 20.
c) Food consumption of F0 females, which gave birth to a litter was determined on postnatal day 1
and 4.

WATER CONSUMPTION: not examined

OTHER: Haematology, clinical chemistry, urinalysis, neurobehavioural examination

Oestrous cyclicity (parental animals):

A vaginal smear was prepared after each mating and examined for the presence of sperm.

Sperm parameters (parental animals):

Parameters examined in male parental generations:
testis weight, epididymis weight

Litter observations:

STANDARDISATION OF LITTERS
- The pups were sexed and examined for macroscopically evident changes on postnatal day 0.
- They were weighed on postnatal day 1 and on postnatal day 4.

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

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):

SACRIFICE
- The female animals were sacrificed 49 days after the beginning of the administration.
- The male animals were sacrificed 34 days after the beginning of the administration.

GROSS NECROPSY
- adrenal glands, aorta, bone marrow (femur), brain, cecum, cervix, coagulating gland, colon, duodenum, eyes with optic nerve, esophagus, extraorbital lacrimal glands, epididymides, femur with knee joint, heart, ileum, jejunum, kidneys, larynx, liver, lungs, lymph nodes, mammary gland, nose, ovaries, oviducts, pancreas, parathyroid glands, pharynx, pituitary gland, prostate gland, rectum, salivary glands, sciatic nerve, seminal vesicles, skeletal muscle, spinal cord, spleen, sternum with marrow, stomach, testes, thymus, thyroid, glands, trachea, urinary bladder, uterus, vagina

HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were prepared for microscopic examination: adrenal glands, all gross lesions, bone marrow (femur), brain, cecum, cervix, coagulating glands, colon, duodenum, epididymes, heart, ileum, jejunum, kidneys, liver, lungs, lymph nodes, ovaries, oviducts, prostate gland, peyer's patches, rectum, sciatic nerve, seminal vesicles. The following organs were weighed: Epididymides, testes (in all animals sacrificed); adrenal glands, brain, heart, kidneys, liver, spleen, thymus (in 5 animals per sex/test group, females with litters only, same animals as used for clinical pathological examinations)

Postmortem examinations (offspring):

SACRIFICE
- Pups were sacrificed under isoflurane anesthesia with CO2 on day 4 of parturition.

NECROPSY
- All pups were examined externally and eviscerated; their organs were assessed macroscopically.

Statistics:

The following statistical tests were used (depending on the parameter): DUNNETT-test, KRUSKAL-WALLIS test, WILCOXON-test (one-side)

Reproductive indices:

male mating index (%), male fertility index (%), female mating index (%), female fertility index (%), gestation index (%), postimplantation loss (%)

Offspring viability indices:

Viability index (%), live birth 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:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Other effects:

not examined

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS):
No clinical signs or changes of general behavior, which may be attributed to the test substance, were detected in any male or female. Several male and female animals of dose group 3 (1000 mg/kg bw/d) showed salivation after treatment. This transient salivation for a few minutes immediately after each treatment was likely to be induced by the unpleasant taste of the test substance or by local irritation of the upper digestive tract. It is not considered to be a sign of systemic toxicity. There were no substance-related mortalities in any of the male and female parental animals in any of the groups. On premating day 12, one high dose F0 female (No. 137 - 1000 mg/kg body weight/day) was found dead after a gavage error.
BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS): Not affected.The fertility index varied between 88.9 % in test groups 3 and 1 and 100% in test group 2 and in control. These values reflect the normal range of biological variation inherent in the strain of rats used for this study. The gestation index was 100% in control and all dose groups.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS): Not affected. The male fertility index ranged between 80% and 100% without showing any relation to dosing.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS): No test substance-related adverse findings

ORGAN WEIGHTS (PARENTAL ANIMALS): When compared to the control group 0 (set to 100%), the mean relative and absolute liver weight was significantly increased in male animals of the high-dose group. All other mean relative and absolute weight parameters in males and all weight parameters in females did not show significant differences when compared to the control group 0. The increase in absolute and relative liver weights in test group 3 (1000 mg/kg bw/day) in male animals was considered to be treatment - related.

GROSS PATHOLOGY (PARENTAL ANIMALS): One female animal that died showed a ruptured esophagus. All other findings occurred individually. They were considered to be incidental or spontaneous in origin and without any relation to treatment.

HISTOPATHOLOGY (PARENTAL ANIMALS): One animal showed rupture of the esophagus confirming the macroscopic diagnosis. All other findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: reproductive performance and fertility

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: general, systemic toxicity

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

VIABILITY (OFFSPRING): The viability index indicating pup mortality during lactation (postnatal day 0 - 4) varied between 86.4% (1000 mg/kg bw/d), 100% (100 and 300 mg/kg bw/d) and 99.2% (control) without showing any association to the treatment.

CLINICAL SIGNS (OFFSPRING): There were no test substance-related adverse clinical signs observed in any of the F1 generation pups of the different test groups.

BODY WEIGHT (OFFSPRING): No test compound-related influence on F1 pup body weights and pup body weight change were noted in all dose groups. Two male runts were seen in test group 3 (1000 mg/kg bw/d). This incidence was considered as spontaneous in nature and not treatment related.

SEX DISTRIBUTION (OFFSPRING): The sex distribution and sex ratios of live F1 pups on the day of birth and postnatal day 4 did not show substantial differences between the control and the test substance-treated groups; slight differences were regarded to be spontaneous in nature.

GROSS PATHOLOGY (OFFSPRING): A few pups showed spontaneous findings at gross necropsy, such as post mortem autolysis, situs inversus, reddish discolored testis, hydroureter and hydronephrosis. Thus, all these findings were not considered to be associated to the test substance.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: developmental toxicity

Reproductive effects observed:

not specified

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

GLP-conform study according to OECD guideline. The test susbstance analogue was investigated.

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 an OECD 422 study, the test substance analogue 3-Methyl-1-vinyl-1 H-imidazolium methyl sulfate was administered daily by gavage to groups of 10 male and 10 female Wistar rats to screen for potential repeated dose, reproductive and developmental toxicity. The animals were dosed throughout the study. For females this included two weeks prior to mating, the variable time of conception, the duration of pregnancy and at least four days after delivery. Males were dosed for a minimum of four weeks. Thereby, both sexes were exposed up to and including the day before scheduled kill with 100, 300, and 1000 mg/kg bw/d. After 2 weeks of premating treatment the F0 animals were mated to produce F1 generation pups. Mating pairs were from the same test group. Mating was discontinued as soon as sperm was detected in the vaginal smear. F0 animals were examined for their reproductive performance including determination of the number of implantation sites and the calculation of post-implantation loss for all F0 females.

With regard to the reproductive performance, the male fertility index ranged between 80% and 100% without showing any relation to dosing. This reflects the normal range of biological variation inherent in the strain of rats used for this study. The fertility index varied between 88.9 % in test groups 3 and 1 and 100% in test group 2 and in control. These values reflect the normal range of biological variation inherent in the strain of rats used for this study. The gestation index was 100% in control and all dose groups. Implantation was not affected by the treatment since the mean number of implantation sites was comparable between all test substance-treated groups and the controls. There were no biologically significant differences in post-implantation loss between the groups and the mean number of F1 pups delivered per dam remained unaffected. The rate of liveborn pups was also not affected by the test substance, as indicated by live birth indices of 100% (control), 98.9 (100 mg/kg bw/d), 98.4 (300 mg/kg bw/d) and 88.5% (1000 mg/kg bw/d).

Concerning the F1 generation, the viability index indicating pup mortality during lactation (postnatal day 0 - 4) varied between 86.4% (1000 mg/kg bw/d), 100% (300 mg/kg bw/d and 100 mg/kg bw/d) and 99.2% (control) without showing any association to the treatment. The pups were sexed and examined for macroscopically evident changes on postnatal day 0. They were weighed on postnatal day 1 and on postnatal day 4. Their viability was recorded. At necropsy on postnatal day 4, all pups were sacrificed with CO2, under isoflurane anesthesia, and examined macroscopically for external and visceral findings. Concerning clinical examinations and gross findings no test substance-related adverse findings were observed in F1 pups.

The NOAEL for reproductive performance and fertility was1000 mg/kg bw/d for the F0 parental rats. The NOAEL for developmental toxicity in the F1 offspring was1000 mg/kg w/d.

 

Short description of key information:
No treatment related parental, reproduction and developmental toxicity was observed at any dose level (100, 300 and 1000 mg/kg bw/day) in an OECD 422 study. Thus, a parental, reproduction and developmental NOAEL of at least 1000 mg/kg bw/d was derived.

Justification for selection of Effect on fertility via oral route:
Only one reliable study.

Effects on developmental toxicity

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available experimental test data is reliable and suitable for the purpose of classification under Directive 67/548/EEC. Based on the data, classification for reproduction/developmental toxicity is not warranted under Directive 67/548/EEC, as amended for the 31st time in Directive 2009/2/EG.  

 

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data is reliable and suitable for the purpose of classification under Regulation (EC) No 1272/2008. Based on the data, classification for reproduction/developmental toxicity is not warranted under Regulation (EC) No 1272/2008, as amended for the sixth time in Regulation No 605/2014.