[2-[[2-cyano-3-[4-(diethylamino)phenyl]-1-oxoallyl]oxy]ethyl][3-[[2-cyano-3-[4-(diethylamino)phenyl]-1-oxoallyl]oxy]propyl]dimethylammonium chloride

Endpoint summary

Description of key information

In an acute oral toxicity study with a dye preparation containing 50% dye the LD50 value was determined to be 2700 mg/kg bw. In an acute dermal toxicity study the LD50 value of the dye was > 2000 mg/kg bw. The intraperitoneal injection with a dye preparation containing 50% dye indicated a LD50 value between 50 and 200 mg/kg bw.

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions (non-GLP)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

Remarks:

pre-GLP

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: WIGA & Hagemann
- Weight at study initiation: males 150 - 270 g, females 150 - 200 g
- Fasting period before study: 15 h - 20 h before administration
- Diet: Herilan MRH-Haltung; H. Eggersmann KG

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 50%, 31.6%, 26.1%, 21.5%, 14.7%, 10.0%

DOSE VOLUME APPLIED:
10 mL/kg bw

Doses:

5000, 3160, 2610, 2150, 1470, 1000 mg/kg bw

No. of animals per sex per dose:

5 rats

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 2 700 mg/kg bw

Based on:

test mat.

Remarks on result:

other: interpolated value

Mortality:

The mortality incidences (sexes combined) were:
5000 mg/kg bw: 9/10
3160 mg/kg bw: 10/10
2610 mg/kg bw: 3/10
2150 mg/kg bw: 3/10
1470 mg/kg bw: 1/10
1000 mg/kg bw: 1/10

For details, see table below

Clinical signs:

other: 5000 mg/kg bw: dyspnea, apathy, staggering, twitching, urine yellow, piloerection, exophthalmos, fascicular, twitching, compulsive gnawing, shortness of breath, poor general state 3160 mg/kg bw: dyspnea, apathy, abnormal position, staggering, salivation,

Gross pathology:

Animals that died: heart: acute dilatation; acute passive hyperemia; lungs: slightly edematized; liver: claycolored periphery involving about half of the area of the acinus, confluent in some animals; stomach: extensive bloody ulcerations in the glandular stomach (hemorrhagic corrosive gastritis); intestines: atony, diarrheal contents
Sacrificed animals: nothing abnormal found in the organs

Mortality:

Dose (mg/kg bw)	5000	3160	2610	2150	1470	1000
Males: Number of animals	5	5	5	5	5	5
Dead animals after:
1 h	4	4	2	0	1	0
1 d	5	5	2	0	1	0
2 d	5	5	2	0	1	0
7 d	5	5	2	0	1	0
14 d	5	5	2	0	1	0
Females: Number of animals	5	5	5	5	5	5
Dead animals after:
1 h	3	5	1	1	0	0
1 d	4	5	1	1	0	0
2 d	4	5	1	3	0	1
7 d	4	5	1	3	0	1
14 d	4	5	1	3	0	1

 

Mean weight (g):

Dose (mg/kg bw)		5000	3160	2610	2150	1470	1000
Males: Beginning of test		190	270	190	180	180	150
After	2-4 d			198	203	209	164
	7 d			244	241	241	198
	13 d			282	273	277	213
Females: Beginning of test		170	200	180	150	160	160
After	2-4 d	184		190	171	179	179
	7 d	210		214	187	194	193
	13 d	229		235	198	204	197

 

LD50 determination:

Doses (mg/kg bw)	No. of animals	Dead animals after 14 days	Mortality (%)	Doses used for calculation
1000	10	1	10
1470	10	1	10
2150	10	3	30
2610	10	3	30	*
3160	10	10	100	*
5000	10	9	90
LD50 about 2700 mg/kg bw (interpolated value)

 

Interpretation of results:

sligthly toxic

Remarks:

Migrated information

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 700 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:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Sulzfeld, Germany
- Age at study initiation: Young adult animals (male animals approx. 8 weeks old, female animals approx. 12 weeks old)
- Weight at study initiation: Animals of comparable weight (mean ± SD: males 233 ± 10 g, females 203 ± 9.8 g)
- Housing: single housing in Makrolon cages, type III
- Diet: VRF1(P) (SDS Special Diets Services, Altrip, Germany)
- Water: Tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12 / 12 (6.00 p.m. - 6.00 a.m. / 6.00 a.m. - 6.00 p.m.)

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: about 40 cm²
- % coverage: at least 10% of the body surface area
- Type of wrap if used: 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)

REMOVAL OF TEST SUBSTANCE
- Washing: rinsing of the application site with warm water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 1.75 mL/kg bw
- Concentration: undiluted

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 rats

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: A check for any dead or moribund animals was made at least once each workday. Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. Individual body weights were determined shortly before administration (day 0), weekly thereafter and on the last day of observation.
- Necropsy of survivors performed: yes
- Other examinations performed: Scoring of skin findings (assessment according to Draize; individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), several times until the last day of observation).

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality and no systemic clinical signs

Mortality:

No mortality occurred.

Clinical signs:

other: No systemic clinical signs were observed during clinical examination. Local effects: Due to the yellowish discoloration of the application area readings of skin redness could not be determined from study day 1 until study day 9 in all male animals, and fr

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:

practically nontoxic

Remarks:

Migrated information

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

Acute oral toxicity

In the key acute oral toxicity study (BASF AG, 1980, equivalent to OECD 401), groups of 5 Sprague-Dawley rats per dose level and sex were exposed by single oral gavage at dose levels of 1000, 1470, 2150, 2610, 3160 and 5000 mg/kg bw with a dye preparation (containing 50% dye and 50% acetic acid). Animals were observed for 14 days. Mortality incidence, clinical signs and body weights were assessed during the observation period. All survivors were necropsied at the end of the observation period and examined macroscopically. Oral administration of the dye preparartion resulted in 1/10, 1/10, 3/10, 3/10, 10/10 and 9/10 deaths at 1000, 1470, 2150, 2610, 3160 and 5000 mg/kg bw, respectively.

As general clinical signs dyspnea, apathy, urine yellow, shortness of breath and poor general state were noted for all groups. All animals surviving to the day of scheduled necropsy showed body weight gain. At necropsy, there were no macroscopic findings in survivors. The oral LD50 for the dye preparation was determined to be 2700 mg/kg bw (interpolated value) in male and female rats.

Acute dermal toxicity

In an acute dermal toxicity study (BASF SE/Bioassay GmbH, 2011, according to OECD 402) the potential of the test substance to exert systemic toxicity was examined in groups of 5 Wistar rats per sex that were semiocclusively exposed for 24 hours and then observed for 14 days. Mortality incidence, clinical signs and body weights were recorded at regular intervals. All survivors were necropsied and examined macroscopically at the end of the observation period. No animal died and no local or systemic clinical signs were observed. There was no test item related effect on body weight. No gross lesions were noted at necropsy. The identified LD50 value was > 2000 mg/kg bw in male and female rats.

 

Other route

In an acute toxicity study (BASF AG, 1980) a dye preparation (containing 50% dye and 50% acetic acid) was administered to 5 mice per sex via intraperitoneal injection at dose levels of 50, 200 or 700 mg/kg bw. Observation period was until day 14 following treatment. All animals of the mid and high dose group died. Clinical signs were observed like dyspnoe, apathy, staggering, tremor, exsiccosis twitching, convulsions, exophthalmos, shortness of breath and a poor general state. Gross pathology revealed staining of abdominal cavity in animals that died and slight intraabdominal agglutination in sacrificed animals. Based on the mortality incidences of 0/10, 10/10 and 10/10 noted at 50, 200 and 700 mg/kg bw, respectively, the LD50 for the i.p. route of exposure was considered to be between 50 and 200 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint
only available reliable study

Justification for selection of acute toxicity – dermal endpoint
only available reliable study

Justification for classification or non-classification

Based on the results of all relevant acute toxicity studies the test item does not need to be subjected to classification and labelling according to Directive 67/548/EEC (DSD) and Regulation (EC) No 1272/2008 (CLP/GHS).