Reaction mass of sodium 3-{[3-(diethylamino)propyl]carbamoyl}-2-(dodecylamino)propanoate and sodium 3-{[3-(diethylamino)propyl]carbamoyl}-3-(dodecylamino)propanoate

Endpoint summary

Description of key information

- 300 mg/kg < Oral LD50 CHIMEXANE HB < 2000 mg/kg

- Dermal LD50 CHIMEXANE HB> 2000 mg/kg

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:

01 March - 19 April 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP study conducted in compliance with OECD guideline 420 with a minor deviation that does not affect study reliability: no rationale was reported for testing additional animals at 2000 mg/kg bw, despite the mortality noted with the first animal in the sighting test.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

yes

Remarks:

no rationale was provided to test other animals at 2000 mg/kg bw, despite the death of the animal tested in the sighting test at this dose.

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

yes

Remarks:

no rationale was provided to test other animals at 2000 mg/kg bw, despite the death of the animal tested in the sighting test at this dose.

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: 8 weeks
- Weight at study initiation: 198 ± 8 g
- Fasting period before study: Animals were fasted for an overnight period of approximately 18 h before dosing, but had free access to water. Food was given back approximately 4 h after administration of the test item.
- Housing: Animals were housed in polycarbonate cages with stainless steel lid. Each cage contained 1-7 rats/sex during the acclimation period and 1 rat (sighting test) and 3 or 5 rats/sex/group (main test) during the treatment period.
- Diet (e.g. ad libitum): SsniffR/M-H pelleted diet (SSNIFF Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water (e.g. ad libitum): Drinking water filtered by a FG Millipore membrane (0.22 µ), ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 30-70 %
- Air changes: Approximately 12 cycles/h of filtered, non-recycled air
- Photoperiod: 12 h dark / 12 h light

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

- Amount of vehicle (if gavage): 10 mL/kg bw
- Rationale for the selection of the starting dose: A preliminary test (sighting test) preceded the main test in which the test item was administered at the dose level of 2000 mg/kg bw to one female. Despite the mortality noted with this first animal, a second female was treated at the concentration of 2000 mg/kg bw. As no mortality occurred, the test item was administered at 2000 mg/kg bw in the main test to three additional females.

Doses:

- Sighting test: 2000 mg/kg bw
- Main test: 300 and 2000 mg/kg bw

No. of animals per sex per dose:

- Sighting test: 2 females
- Main test: 3 or 5 females

Control animals:

other: historical control data

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical signs: Animals were observed frequently during the hours following administration of the test item; subsequently once daily up to 14 days
Body weight was recorded on Days 1 (just before administration), 8 and 15.
- Necropsy of survivors performed: Yes, all surviving animals were killed by carbon dioxide asphyxiation on Day 15.
- All animals were subjected to a macroscopic examination which consisted of opening the thoracic and abdominal cavities. The macroscopic appearance of all examined organs was recorded.

Statistics:

None

Preliminary study:

In the sighting test, the test item was first administered at the dose level of 2000 mg/kg bw to one female. Despite the mortality noted with this first animal, a second female was treated at the concentration of 2000 mg/kg bw. As no mortality occurred, the test item was administered at the dose-level of 2000 mg/kg bw in the main test to three additional females.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

- At 2000 mg/kg bw: 2/5 females were found dead on Day 2 or 3.
- At 300 mg/kg bw: No mortality was noted.

Clinical signs:

other: - At 2000 mg/kg bw: Hypoactivity, piloerection and dyspnea were noted in 2/5 females prior to the death, from Day 1. Soft faeces were also noted in one female on Day 1 only. Piloerection, concurrently with hypoactivity and/or dyspnea were observed in surv

Gross pathology:

- No macroscopic abnormalities were observed at necropsy.

Other findings:

None

None

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The oral LD50 for CHIMEXANE HB is greater than 300 mg/kg bw and less than 2000 mg/kg bw in rats therefore it is classified as ‘R22 Harmful if swallowed’ according to the Annex VI to the Directive 67/548/EEC and ‘Category 4’ according to the CLP Regulation (EC) N° (1272-2008).

Executive summary:

In a GLP acute oral toxicity study performed according to OECD Guideline 420, groups of female Sprague Dawley [Rj: SD (IOPS Han)] rats were given a single oral dose of CHIMEXANE HB at 300 and 2000 mg/kg bw. A preliminary test (sighting test) preceded the main test in which the test item was administered at the dose level of 2000 mg/kg bw to one female. Despite the mortality noted with this first animal, a second female was treated at the concentration of 2000 mg/kg bw. As no mortality occurred, the test item was administered at 2000 mg/kg bw in the main test to three additional females. All animals were observed for mortality, clinical signs and bodyweights for 14 days and were all sacrificed for macroscopic examination.

 

At 2000 mg/kg bw, 2/5 females were found dead on Day 2 or 3. Hypoactivity, piloerection and dyspnea were noted prior to the death, from Day 1. Soft faeces were also noted in one of them on Day 1 only. Piloerection, concurrently with hypoactivity and/or dyspnea were observed in surviving animals between Days 1 and 3. At 300 mg/kg bw, no mortality and no clinical signs were observed. When compared to historical control animals, a slightly reduced body weight gain was noted in 1/5 and 2/5 females at 300 and 2000 mg/kg bw, respectively. The overall body weight gain of the other treated animals was not affected by treatment with the test item. No macroscopic abnormalities were observed at study termination on Day 15. In this study, the oral LD50 of CHIMEXANE HB was considered to be 300-2000 mg/kg bw in female rats.

 

The oral LD50 for CHIMEXANE HB is greater than 300 mg/kg bw and less than 2000 mg/kg bw in rats therefore it is classified as ‘R22 Harmful if swallowed’ according to the Annex VI to the Directive 67/548/EEC and ‘Category 4’ according to the CLP Regulation (EC) N° (1272-2008).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

GLP-compliant study with a Klimish 1

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

Study period:

14-30 March 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted in compliance with OECD guideline 402 without any deviations.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

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: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: Approximately 8 weeks old
- Weight at study initiation: Males: 353 ± 12 g; females: 217 ± 9 g
- Housing: Housed individually in polycarbonate cages
- Diet: SsniffR/M-H pelleted diet (SSNIFF Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water: Drinking water filtered by a FG Millipore membrane (0.22 µm), ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 30-70 %
- Air changes: Approximately 12 cycles/h of filtered, non-recycled air
- Photoperiod: 12 h dark / 12 h light

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: Dorsal area, approximately 5 cm x 7 cm for males and 5 cm x 6 cm for females
- % coverage: Approximately 10 % of the total body surface area
- Undiluted test item was placed on a hydrophilic gauze pad and then applied on the clipped skin. The test item and the gauze pad were held in contact with the skin for 24 h by means of an adhesive hypoallergenic aerated semi-occlusive dressing and a restraining bandage.

REMOVAL OF TEST SUBSTANCE
- Washing: Residual test item was removed using a moistened cotton pad.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Constant volume or concentration used: Yes

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

- Sighting test: 1/sex
- Main test: 4/sex

Control animals:

other: historical control data

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical signs and mortality: Animals were observed frequently during the hours following administration of the test item, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day until Day 15. From Day 2, any local cutaneous reaction was recorded.
Bodyweight was recorded on Days 1 (prior to dosing), 8 and 15.
- Necropsy of survivors performed: Yes; all animals were killed by carbon dioxide asphyxiation on Day 15 and all animals were subjected to a macroscopic examination.

Statistics:

None

Preliminary study:

Not applicable

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed.

Clinical signs:

other: No clinical signs were observed.

Gross pathology:

- No macroscopic abnormalities were observed at necropsy.

Other findings:

Skin reactions:
- An erythema was observed in all animals from Day 2. This erythema persisted up to Day 4 in 1/5 males, Day 7 in 1/5 males and 1/5 females, Day 9 in 2/5 males and 1/5 females, Day 11 in 1/5 males and Day 12 in 3/5 females.
- An edema was observed in 4/5 males and 4/5 females from Day 2 up to Day 4.
- A dryness of the skin was observed in 4/5 males and 4/5 females from Day 5. It persisted up to Day 12 in 3/5 males and 4/5 females and up to Day 13 in 1/5 males.
- Crusts were noted in 3/5 males and 4/5 females from Day 5. They persisted up to Day 7 in 3/5 males and 2/5 females, Day 9 in 1/5 females and Day 12 in 1/5 females.

None

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the test conditions, CHIMEXANE HB should not be classified according to the criteria of the Annex VI of the Directive 67/548/EEC and the CLP Regulation (EC) N° (1272-2008).

Executive summary:

CHIMEXANE HB was tested for acute dermal toxicity in Sprague-Dawley [Rj: SD (IOPS Han)] rats in a GLP-compliant limit dose assay according to OECD guideline 402. Groups of rats (5/sex) were administered a single dermal dose of undiluted test material at 2000 mg/kg bw on clipped skin (approximately 10 % of the total body surface area) using a semi-occlusive patch held in place for 24 h. Residual test item was removed using a moistened cotton pad at the end of the 24 h exposure period. Examinations for mortality, clinical signs, body weight gain and dermal reactions were performed during a 14-day observation period. All surviving animals were necropsied at the end of the observation period.

 

No deaths and clinical signs occurred during the observation period. When compared to historical control animals, a slightly reduced body weight gain was recorded in 1/5 males between Days 1 and 8 and in 3/5 males and 2/5 females between Days 8 and 15. Overall body weight gain of the other animals was similar to that of historical control animals. At necropsy, macroscopic examination of main organs showed no abnormalities. The acute dermal combined LD50 was greater than 2000 mg/kg bw.

 

Some dermal changes were observed in most of the animals. They consisted of erythema, edema, skin dryness and later crusts. An erythema was observed in all animals from Day 2 up to Day 4 in 1/5 males, Day 7 in 1/5 males and 1/5 females, Day 9 in 2/5 males and 1/5 females, Day 11 in 1/5 males and Day 12 in 3/5 females. An edema was observed in 4/5 males and 4/5 females from Day 2 up to Day 4. A dryness of the skin was observed in 4/5 males and 4/5 females from Day 5 ; it persisted up to Day 12 in 3/5 males and 4/5 females, and up to Day 13 in 1/5 males. Crusts were noted in 3/5 males and 4/5 females from Day 5 up to Day 7 in 3/5 males and 2/5 females, Day 9 in 1/5 females and Day 12 in 1/5 females.

 

Under the test conditions, CHIMEXANE HB should not be classified according to the criteria of the Annex VI of the Directive 67/548/EEC and the CLP Regulation (EC) N° (1272-2008).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

GLP-compliant study with a Klimish 1

Additional information

In a GLP-compliant acute oral toxicity study performed in accordance with OECD guideline 420 in Sprague Dawley rats, a single dose of 2000 mg/kg bw of Chimexane HB induced the death of 2/5 animals. Due to these results, a second group of 5 animals/sex received a single oral dose of 300 mg/kg bw of Chimexane HB. No mortality and no clinical signs were observed in the animals that received this lower dose. Thus, the oral LD50 of Chimexane HB is in the range of 300 -2000 mg/kg bw.

As the submission substance was tested in an aqueous solution (Chimexane HB - submission substance 55.9% in water), the oral LD50 of the submission substance is in the range of 167.7 -1118 mg/kg bw. As mortality was observed in 2/5 animals at the dose equivalent to 1118 mg submission substance/kg bw, the submission substance oral LD50 is closer to 1118 mg/kg bw than to 167.7 mg/kg bw. Thus, submission substance oral LD50 should be considered to be in the range of 300 -2000 mg/kg bw.

In a GLP-compliant acute dermal toxicity study performed in accordance with OECD guideline 402, no mortality was observed in Sprague Dawley rats given a single dose of Chimexane HB at the limit test dose of 2000 mg/kg bw. Thus, Chimexane HB dermal LD50 is > 2000 mg/kg bw, corresponding to 1118 mg/kg bw of the submission substance.

As the oral LD50 of the submission substance is close to 1000 mg/kg bw, and as no further data could be generated, it is therefore conservatively considered that the submission substance dermal LD50 is in the range of 1000 -2000 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint

Only study available

Justification for selection of acute toxicity – inhalation endpoint

No study was available and it was not necessary to provide one because acute oral toxicity was studied for two other routes of administration and the inhalation route of exposure is irrelevant for the submission substance. Acute toxicity studies were performed for the oral and dermal route, and the submission substance showed low toxicity (LD50 between 1000 and 2000 mg/kg bw). Also, as  the submission substance has a low volatility (vapor pressure < 1 Pa), and is manufactured, marketed and used in water (as Chimexane HB),  constituting a gel, exposure to the submission substance by inhalation is not expected.

Justification for selection of acute toxicity – dermal endpoint

Only one study available

Justification for classification or non-classification

As the submission substance oral LD50 are in the range of doses of 300 -2000 mg/kg bw, and as its dermal LD50 is in the range 1000 -2000 mg/kg bw, it should be classified "R21/22 Harmful in contact with skin and if swallowed" according to the Annex VI to the Directive 67/548/EEC, and Category 4 for acute oral and dermal toxicity according to the CLP Regulation (EC) N° 1272/2008, with the hazard statements "H302: Harmful if swallowed" and "H312: Harmful in contact with skin".