1-ethyl-3-methyl-3H-imidazolium dicyanidoamide(1-)

Endpoint summary

Description of key information

The LD50 through oral exposure was >2000 mg/kg bw in a GLP-compliant OECD 423 study.

The LD50 through dermal exposure was >2000 mg/kg bw in a GLP-compliant OECD 402 study.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

- Name of the test substance (as cited in study report): EMIM Dicyanamid
- Purity: 97.4 g/100 g
- Density [g/mL]: 1.099 (determined by Bioassay)

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

- Age/weight: Young adult animals of a comparable weight were used.
- Acclimatization period: at least 5 days before the beginning of the experimental phase; during the acclimatization period, the animals were accustomed to the environmental conditions of the study and to the diet.
- Individual animal identification: by cage cards and tail marking.
- Conditions: the animals were housed in fully air-conditioned rooms. Central air-conditioning guaranteed a range of 22°C ± 3°C for temperature and of 30 – 70% for relative humidity. The day/night rhythm was 12 h light and 12 h darkness.
- Housing: single housing in Makrolon cages, type III.
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany. Feed was withdrawn from the animals at least 16 hours before administration.
- Water: Tap water ad libitum

Route of administration:

oral: capsule

Vehicle:

other: unchanged and water

Details on oral exposure:

For the high dose, the liquid test item was administered unchanged. For the lower dose, an administration volume of 2 mL/kg bw of suitable test item preparations was used to facilitate application. The test item preparation was produced for the 500 mg/kg application group shortly before application by stirring with a magnetic stirrer. Concentration used: 25 g/100 mL (500 mg/kg dose level). Form of administration: solution.

Doses:

500 and 2000 mg/kg

No. of animals per sex per dose:

- 500 mg/kg: 3
- 2000 mg/kg: 6

Control animals:

no

Details on study design:

- Observation period: 14 days
- Individual body weight determination shortly before administration (day 0), weekly thereafter and on the last day of observation.
- Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals.
- A check for any dead or moribund animals was made at least once each workday.
- Necropsy with gross-pathology examination on the last day of the observation period after sacrifice by CO2 inhalation in a chamber with increasing concentrations over time. Necropsy of all animal that died before as early as possible after death.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

- 2000 mg/kg (first test group): 1/3 animals
- 2000 mg/kg (second test group): no deaths

Clinical signs:

other: - 2000 mg/kg (first test group): Impaired and poor general state, dyspnoea, piloerection, tremor, staggering, abdominal position, tonic convulsions, exsiccosis, reduced feces, none feces - 2000 mg/kg (second test group): impaired and poor general state, d

Gross pathology:

Animal that died: liquid clear content in the stomach, red discoloration of the glandular stomach, dark red discoloration of the liver. There were no macroscopic pathological findings in the surviving animals sacrificed at the end of the observation period.

Interpretation of results:

Category 5 based on GHS criteria

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

February 24, 1987

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

August 1998

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Test Guideline of 12 Nosan No. 8147

GLP compliance:

yes (incl. QA statement)

Remarks:

Bioassay, Labor für biologische Analytik GmbH, 69120 Heidelberg

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

- Name of test material (as cited in study report): EMIM Dicyanamid
- Test item No.: 12/0071-2
- Batch identification: 0013141983
- Content: 97.4 g/100 g determined by 1H-NMR spectroscopy
- Homogeneity: the test item was homogeneous by visual inspection
- Storage stability: the stability of the test item under storage conditions over the study period was guaranteed by the sponsor
- Expiry date: February 19, 2017
- Storage conditions: room temperature, under N2
- Physical state / color: liquid / colorless to brown, clear

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI (Han) SPF

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: young adult animals (male animals approx. 8 weeks, female animals approx. 12 weeks)
- Weight at study initiation: males: 222 - 242 g; females: 207 - 215 g
- Housing: single housing in Makrolon cages type III, including bedding (H 15005-29; Ssniff, Spezialdiäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany) and enrichment (Wooden gnawing blocks (Type NGM E-022) ; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria)
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
- Water: tap water ad libitum
- Acclimation period: acclimatization period of at least 5 days before the beginning of the experimental phase; during the acclimatization period, the animals were accustomed to the environmental conditions of the study and to the diet.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12 h / 12 h (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.)

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE/EXPOSURE
- Clipping of the fur: about 24 hours before application
- Route of application: single application to the clipped epidermis (dorsal and dorsolateral parts of the trunk); covering of the application site with a semi-occlusive dressing (the test item was covered with an air-permeable dressing; 4 layers of absorbent gauze (Ph. Eur. supplied by Lohmann GmbH & Co., KG) and stretch bandage (Fixomull® Stretch (adhesive fleece) supplied by Beiersdorf AG) for 24 hours. Afterwards removal of the semi-occlusive dressing, rinsing of the application site with warm water.
- Application area: about 40 cm² (corresponds to at least 10% of the body surface)
- Time of day of application: in the morning

TEST MATERIAL
- Amount(s) applied: 1.80 mL/kg bw

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Observation period: 14 days
- Body weight determination: individual body weights shortly before application (day 0), weekly thereafter and on the last day of observation.
- Clinical observations: clinical signs for each animal were recorded several times on the day of application and at least once during each workday thereafter.
- Scoring of skin findings: individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), weekly thereafter and on the last day of observation.
- Mortality: a check for any dead or moribund animals was made at least once each workday.
- Pathology: necropsy with gross-pathology examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually increasing concentrations.
- Assessment of skin reactions: the evaluation of skin reactions was performed according to Draize, J. H. "Dermal toxicity." Appraisal of the safety of chemicals in foods, drugs and cosmetics (1959): 46-59. Appraisal of the safety of chemicals in foods, drugs and cosmetics. The association of food and drug officials of the United States Austin, Texas.

Statistics:

Calculations were performed using Microsoft Excel 2003 and checked with a calculator.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No mortality occurred.

Clinical signs:

other: No systemic clinical signs or local effects were observed during clinical examination.

Gross pathology:

No macroscopic pathologic abnormalities were noted in the animals (5 males and 5 females) examined on the last day of observation.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Additional information

Acute oral toxicity was determined in a study according to OECD 423 and in compliance with GLP criteria. In this study, 3 male and 3 female Wistar rats were exposed to the liquid test substance diluted with water in a single dose of 500 mg/kg mg/kg bw (2 mL/kg bw administration volume). Additionally, in total 6 males and 6 females (divided over 2 test groups) were exposed to the liquid undiluted test substance in a single dose of 2000 mg/kg bw. The highest dose was administered orally with a capsule, whereas the lower dose was applied via solution. In addition to daily survival inspections, individual body weights were determined weekly, clinical signs were recorded daily and at the end of the study period of 14 days, necropsy was performed. Overall, in the first test group of the 2000 mg/kg bw dose 1 animal died, whereas in the second group of that dose no deaths were observed. Clinical signs that were noted include impaired and poor general state, dyspnoea, piloerection, tremor, staggering, abdominal position, tonic convulsions, exsiccosis, reduced feces, none feces in the high dose group (2000 mg/kg bw) and impaired general state, dyspnoea and piloerection in the low dose group (500 mg/kg bw). All animals gained weight during the study period with the exception of one animal of both 2000 mg/kg dose groups respectively, which showed stagnation of body weights either during the first post-exposure week or during the second post-exposure week. The animal that died showed liquid clear content in the stomach, red discoloration of the glandular stomach and dark red discoloration of the liver. There were no macroscopic pathological findings in the surviving animals sacrificed at the end of the observation period. Based on these results, the LD50 was determined at >2000 mg/kg bw.

Furthermore, the acute dermal toxicity was determined in a limit test according to OECD 402 and in compliance with GLP criteria. In this study, 5 male and 5 female Wistar rats were exposed to undiluted test substance in a dose of 2000 mg/kg bw (1.80 mL/kg bw) by single application to the clipped epidermis, under semi-occlusive conditions for 24 hours. After, the application site was rinsed with warm water. Animals were observed for a period of 14 days. Clinical observations and survival were determined daily, whereas skin findings (scored according to Draize) and body weight were determined weekly. At the end of the study period, gross pathology was performed. Overall, no mortality occurred and no systemic clinical signs or local effects were observed during clinical examination. All animals gained weight during the study, although 2 females stagnated in weight gain in the first week; the body weight stagnation observed is considered to be unspecific. Based on these results, the LD50 was determined to be >2000 mg/kg bw.

Justification for classification or non-classification

Based on the available information classification for acute oral or dermal toxicity is not warranted in accordance with EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008.

However, according to GHS UN (ST/SG/AC.10/30Rev.6, 2015) , the substance is classified for acute oral toxicity: Acute Tox. Cat. 5; H303: May be harmful if swallowed.