Succinonitrile

Endpoint summary

Description of key information

An inhalation toxicity study and an acute dermal toxicity study were performed in accordance with the current OECD guideline. Both studies are reliable with restrictions.

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

Study period:

2015

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)

Deviations:

yes

Remarks:

Animals where housed with 2 in a cage during acclimatisation. The animals showed normal during the acclimatization period, it was considered not to affect the animals and study results.

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: DSM Engineering Plastics B.V. lotnumber SN0620140520
- Expiration date of the lot/batch: 2016.07.01
- Purity test date: 99.93%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store in a cool area (IO-20°C). Containers that have been opened must be filled with dry nitrogen for at least 1min and then sealed. Be careful not to import any water or impurities during the filling and sealing course.
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: Easily soluble in cold water


FORM AS APPLIED IN THE TEST white waxy solid

Species:

rat

Strain:

other: Sprague Dawley (SD)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Supplier: Beijing Vital River Laboratory Animal Technology Co., Ltd.
License No.: SCXK (Jing) 2012—0001, Qualification: 11400700112907.

- Age at study initiation: 51-60 days on arrival, in the range of 8-12 weeks at the commencement of each
animal’s dosing.
- Weight at study initiation: 182-222g
- Housing:Animals were raised
in suspended, stainless steel cages (L32.0cm >(L167.0cmXW70.0cm>Animals were housed individually during the test.
- Diet : diet with complete nutrition supplied by Beijing keaoxieli Feed Co., Ltd. Product License No: SCXKUing) 2012-0001, Batch NO. 15063223).
- Water (e.g. ad libitum): purified water (by HT-R01000 purity system.)
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.5-24.5 °C
- Humidity (%): 45%-70%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): lighting sequence was 12 hours light, 12 hours dark.

Route of administration:

oral: gavage

Doses:

300 mg/kg BW and 2000 mg/kg BW

No. of animals per sex per dose:

3

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 30 min and at l, 2 and 4 hours after dosing and then once each day for up to 14 days.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems, and somatomotor activity and behaviour pattern. Attention had been directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Preliminary study:

LD50 is 488.7 mg/kg b.w. for rats was available. Based on that value the test doses were selected.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 300 - <= 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Mortality
Dose Level-The first dosing (300mg/kg): One animal was dead on Day 2 after dosing. Dose Level-The second dosing (300mg/kg): There were no deaths or moribund during the test. Dose Level-The third dosing (2000mg/kg): All animals were dead in 4h after dosing. Mortality summary results were given in appendix Table 1.

Clinical signs:

other: Dose Level-The first dosing (300mg/kg): Animals showed decrease in locomotor activity, emaciation, and soiled perinea region after dosing. Dose Level-The second dosing (3 00mg/kg): There were no abnormal findings after dosing from the first day until the end

Gross pathology:

Necropsy
No abnormalities were found at necropsy.
Necropsy results see appendix table 4.

Table 1. Mortality summary results
Dosing sequence	Dose (mg/kg)	sex	Animal amounth	Time (d)															Mortality
				0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
1	300	Female	3	0	0	1	0	0	0	0	0	0	0	0	0	0	0	0	1/3
2	300	Female	3	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0/3
3	2000	Female	3	3	0	0	0	0	0	0	0	0	0	0	0	0	0	0	3/3

Table 2. Individual Clinical observations
Dose (mg/kg)	Animal ID	Symptons After Dosing (Hours)				Symptoms During Period After Dosing (Days)
		0.5	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
300	2100	N	N	N	N	S	E1D	-	-	-	-	-	-	-	-	-	-	-	-
	2101	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N
	2102	N	N	N	N	H	N	N	N	N	N	N	N	N	N	N	N	N	N
300	2200	N	N	N	N	N	E1D	-	-	-	-	-	-	-	-	-	-	-	-
	2201	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N
	2202	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N	N
300	2300	N	SE	SE Se	FD	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	2301	N	SE	SE SeW	FD	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	2302	N	N	SE	FD	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Note: N=no sign; S=decrease in locomotor activity; E1=emaciation, E=secretion around mouth and nouse, Se-semi closed eyes, W=weak breath, D=death, F=soiled fur around the mouth and nouse, H=soiled perinea region.

Table 3. Individual and Mean Body Weights
Dose (mg/kg)	Dose (mg/kg)	Animal ID	Body Weight (g) at Day					Body Weight Gain (g) During Week
			Grouping	Day0	Day7	Day14	At Death	1	2
300	Female	2100	204	191	-	-	149	-	-
		2101	233	212	246	268	-	34	22
		2102	226	209	255	263	-	46	8
		Mean +/- SD	221 +/- 15	204 +/- 11	251 +/- 6	266 +/- 4	-	40 +/- 8	15 +/- 10
		N	3	3	2	2	1	2	2
300	Female	2200	201	183	-	-	149	-	-
		2201	219	212	246	268	-	34	22
		2202	225	209	255	263	-	46	8
		Mean +/- SD	215+/- 12	199 +/- 15	245 +/- 16	226 +/- 11	-	46 +/- 9	-20+/-6
		N	3	3	3	3	-	3	3
2000	Female	2300	196	191	-	-	184.53	-	-
		2301	216	212	-	-	199.86	-	-
		2302	222	209	-	-	203.18	-	-
		Mean +/- SD	211 +/- 14	198 +/- 7	-	-	-	-	-
		N	3	3	0	0	3	2	2
Note: body weight(g), N=Number of animals per group, SD= Standard deviation

Table 4. Indivudual Negroscopy Findings
Animal ID	sex	Dose (mg/kg)	Death time	Findings
2100	Female	300	Day2	No abnormalities
2101	Female	300	Sheduled Negroscopy	No abnormalities
2102	Female	300	Sheduled Negroscopy	No abnormalities
2200	Female	300	Sheduled Negroscopy	No abnormalities
2201	Female	300	Sheduled Negroscopy	No abnormalities
2202	Female	300	Sheduled Negroscopy	No abnormalities
2300	Female	2000	Day 0	No abnormalities
2301	Female	2000	Day 0	No abnormalities
2302	Female	2000	Day 0	No abnormalities

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Based on the results, the acute oral LDso in the rats for Succinonitrile was estimated to be among 300 mg/kg b.w. - 2000 mg/kg b.w..
According to the GHS’s classification criteria for acute oral toxicity, the test item was classified as Category 4.

Executive summary:

Introduction: The study was performed to assess the acute oral toxicity of Succinonitrile in Sprague Dawley rats. The method was designed to meet the OECD Guideline for Testing of Chemicals: Acute Oral Toxicity-Acute Toxic Class Method (TG423, 2001).

Method: The test item was tested using a stepwise procedure, each group using three female animals. The first step dosing and second step dosing were 300 mg/kg and the third step dosing was ZOOOmg/kg. Clinical observations and bodyweight were monitored during the study. All animals were subjected to a gross necropsy.

Results:

Mortality

Dose Level-The first dosing (300mg/kg): One animal was dead on Day 2 after dosing.

Dose Level-The second dosing (300mg/kg): There were no deaths or moribund during

the test.

Dose Level-The third dosing (2000mg/kg): All animals were dead in 4h after dosing.

Clinical observations

Dose Level-The first dosing (300mg/kg): Animals showed decrease in locomotor

activity, emaciation, and soiled perinea region after dosing.

Dose Level-The second dosing (3 00mg/kg): There were no abnormal findings after

dosing from the first day until the end of the test.

Dose Level-The third dosing (2000mg/kg): Animals showed decrease in locomotor

activity, emaciation, secretion around mouth and nose, semi-closed eyes, weak breath,

soiled fur around the mouth and nose after dosing.

Body Weight

All surviving animals showed a increase in body weight during the first week, but most of the surviving animals showed a decrease during the second week.

Necropsy

No abnormalities were found at necropsy.

Conclusion:

Based on the results, the acute oral LDso in the rats for Succinonitrile was estimated to be among 300 mg/kg b.w. - 2000 mg/kg b.w.. According to the GHS’s classification criteria for acute oral toxicity, the test item was classified as, Category 4.

Endpoint conclusion

Dose descriptor:

LD50

Value:

300 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Dose descriptor:

discriminating conc.

Value:

2 670 mg/m³ air

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

Study period:

2015

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:

Acute Dermal Toxicity (TG402, adopted 1987).

GLP compliance:

yes (incl. QA statement)

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: DSM Engineering Plastics B.V. lotnumber SN0620140520
- Expiration date of the lot/batch: 2016.07.01
- Purity test date: 99.93%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store in a cool area (IO-20°C). Containers that have been opened must be filled with dry nitrogen for at least 1min and then sealed. Be careful not to import any water or impurities during the filling and sealing course.
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: Easily soluble in cold water


FORM AS APPLIED IN THE TEST white waxy solid

Species:

rat

Strain:

other: Sprague Dawley (SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 56-65 days on arrival, in the range of 8-12 weeks at the commencement of each animal’s dosing.
- Weight at study initiation: Body weight range: Males: 23 5-270g. Females: 225-235g, +/- 20% of the mean body weight at grouping.
-
- Housing: Animals were housed individually during the test.
- Diet : provided sterilized diet with complete nutrition supplied by Beijing keaoxieli Feed
- Water : Purified water
- Acclimation period: 5 days prior to the test

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.0-24.5"C
- Humidity (%): 45%-70%
- Photoperiod (hrs dark / hrs light): light cycle was 12 hour light/ 12 hour dark.

Type of coverage:

not specified

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: fur removed from dorsal area by shaving, clear area 40cm2. A gauze 5cm x 5.50m is used

- Type of wrap if used: small piece of white medical tape(non-irritating)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): esidual test item was
removed by cotton wool soaked in water.
- Time after start of exposure: approximated 24 hour

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg
- For solids, paste formed: yes

VEHICLE
- Amount(s) applied (volume or weight with unit): test item was moistened with the vehicle
- Purity: purified water

Duration of exposure:

Approximately 24 hours

Doses:

2000

No. of animals per sex per dose:

5 Male
5 female

Control animals:

no

Details on study design:

- Duration of observation period following administration: Clinical observations were made during the first 30 min, and at 1, 2 and 4 hours after dosing and then once each day for up to 14 days.
- Frequency of observations and weighing: ndividual weights of animals were determined at grouping, Day 0(day of dosing), Day7 and Dayl4
- Necropsy of survivors performed: yes
- Other examinations performed: Careful observation and record of animal fur changes, eyes and mucosa, respiratory, circulatory, nervous system, particularly limb activity and behavior change were made. Particular attention was directed to observations of tremor, convulsions, salivation, diarrhea, lethargy, sleep and coma.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths or moribund during the test.

Clinical signs:

other: There were no abnormal findings in all animals after dosing from the first day until the end of the test.

Gross pathology:

No abnormalities were found at necropsy

Other findings:

Skin Reactions: There were no signs of dermal irritation.

Interpretation of results:

other: Category 5 or unclassified.

Conclusions:

Conclusion: Based on the results, the acute dermal LD50 in rats for Succinonitrile was as follows:
Male rats :>2000mg/kg b.w.
Female rats :>2000mg/kg b.w.
According to the GHS’s classification criteria of acute dermal toxicity, the test item was classified as Category 5 or unclassified.

Executive summary:

Introduction: The study was performed to assess the acute dermal toxicity of

Succinonitrile in Sprague Dawley rats. The method was designed to meet the OECD

Guideline for Testing of Chemicals: Acute Dermal Toxicity (TG402, adopted 1987).

Method: A limit test at one dose level of 2000mg/kg body weight was carried out in a

group of 10 animals (5 males and 5 females). Clinical observations were made during

the first 30 min, and at 1, 2 and 4 hours after dosing and then once each day for up to 14

days. Individual weights of animals were determined at grouping, Day 0(day of dosing),

Day7 and Dayl4. At the end of the test, a gross necropsy was performed on all animals

under test.

Results:

Mortality

There were no deaths or moribund during the test.

Clinical observations

There were no abnormal findings in all animals after dosing from the first day until the

end of the test.

Skin Reactions

There were no signs of dermal irritation.

Body Weight

One male and three female animals showed a decrease in body weight during the first

week. But overall also these animals showed growth during the second week. The

results indicated that the body weight gain of animals showed growth.

Necropsy

No abnormalities were found at necropsy.

Conclusion: Based on the results, the acute dermal LD50 in rats for Succinonitrile was

as follows:

Male rats :>2000mg/kg b.w.

Female rats :>2000mg/kg b.w.

According to the GHS’s classification criteria of acute dermal toxicity, the test item was

classified as Category 5 or unclassified.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

An inhalation toxicity study was performed with 5 rats/sex in accordance with the current OECD guideline. The rats were exposed whole body to the substance by aerosols at the maximal attainable concentration. None of the animals died during the exposure period, but three males died within 48 hours after termination of the exposure. It was concluded that the 4-hour LC50 was higher than 2.67 mg/l of air. Since the LC50 was found to be higher than the highest attainable concentration, the substance does not have to be classified for inhalation toxicity according to CLP Regulation (EC) 1272/2008.

Acute dermal toxicity study with Sprague Dawley:

A limit test at one dose level of 2000mg/kg body weight was carried out in a group of 10 animals (5 males and 5 females). Clinical observations were made during the first 30 min, and at 1, 2 and 4 hours after dosing and then once each day for up to 14

days. Individual weights of animals were determined at grouping, Day 0(day of dosing), Day7 and Dayl4. At the end of the test, a gross necropsy was performed on all animals under test.

Based on the results, the acute dermal LD50 in rats for Succinonitrile was as follows:

Male rats :>2000mg/kg b.w.

Female rats :>2000mg/kg b.w.

According to the CLP Regulation (EC) 1272/2008 the test item is unclassified .

Justification for classification or non-classification

Based on the above inhalation toxicity study, the substance does not have to be classified for inhalation toxicity.

Based on the above acute dermal toxicity study, the substance has to be classified for acute dermal toxicity in category 3 according to CLP Regulation (EC) No 1272/2008.