Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: Test method OECD 423, GLP study. The acute oral LD50 was found to be above 2000 mg/kg bw in female rats.

Acute inhalation toxicity: Acute toxicity information provided for oral and dermal route.

Acute dermal toxicity: Test method OECD 402, GLP study. The acute dermal LD50 was determined to be > 2000 mg/kg bw in male and female rats. Severe skin reaction were observed.

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:
23 April 2013 - 8 May 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test method according to OECD 423. GLP study.
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
other: CRL:(WI)
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld
- Age at study initiation: 8 weeks old
- Weight at study initiation: 196–209 g
- Fasting period before study: Yes, the night before administration.
- Housing: 3 animals/cage. Type II polypropylene/polycarbonate cage with lignoce bedding
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5 – 23.3 °C
- Humidity (%): 34 – 68 %
- Air changes (per hr): 15 – 20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark
Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Lot/batch no. (if required): BCBH5068V

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg bw (10 mL/kg bw)

DOSAGE PREPARATION (if unusual): The test item was freshly formulated at a concentration of 200 mg/mL in the vehicle. The formulation container was stirred continuously up to finishing the treatment.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: In the lack of any preliminary toxicological information, 2000 mg/kg bw was selected to be the starting dose.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6 animals, 3 animals/group
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: See below.
- Necropsy of survivors performed: yes
- Examinations performed:
Clinical signs: 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Observations: skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Body weight: Recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly thereafter.
Necropsy: External appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.
Statistics:
The method used was not intended to allow the calculation of a precise LD50 value.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortalities were observed.
Clinical signs:
The test substance did not cause any clinical signs during the 14 days observation period.
Body weight:
Body weight gains during the study showed no indication of a test item-related effect.
Gross pathology:
No macroscopic observations were seen in animals terminated on Day 14.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute oral LD50 value of the test item was found to be above 2000 mg/kg bw in female CRL:(WI) rats.
Executive summary:

The single-dose oral toxicity of the substance was performed according to the acute toxic class method OECD 423 in CRL: (WI) rats. Initially, three females were treated at a dose level of 2000 mg/kg bw. As no mortality was observed, a confirmatory group was treated at the same dose level. No mortality was observed in the confirmatory group, therefore no further testing was required. Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination. No test item related effects were observed during the study performance. The acute oral LD50 value was found to be above 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 001 mg/kg bw
Quality of whole database:
The study is a GLP compliant and has Klimisch score 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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to OECD Guidelines and to GLP.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
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: Harlan Laboratories UK Limited
- Age at study initiation: 8-12 w eeks
- Weight at study initiation: >200 g
- Housing: individually in suspended solid floor polypropylene cages
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25°C
- Humidity (%): 30-70%
- Air changes (per hr): 15 changes/hour
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- % coverage: 10%
- Type of wrap if used: surgical gauze

REMOVAL OF TEST SUBSTANCE
- Washing (if done): removal w ith cotton w ool moistened w ith arachis oil
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or w eight w ith unit): 2.16 ml/kg
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period follow ing administration: 14 days
- Frequency of observations and w eighing: 30 minutes, 1, 2 and 4 hours, once daily for fourteen days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body w eight, gross lesions
Statistics:
Use of statistics not indicated.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
Deaths did not occur during the study.
Clinical signs:
Systemic toxicity was not observed.
Body weight:
Three females showed bodyw eight decrease or no gain in bodyw eight during the first w eek with expected gain in bodyweight during the second week. One female showed expected gain in bodyweight during the first week w ith bodyweight decrease during the second week. All males and one female show ed expected gains in bodyweight over the study period.
Gross pathology:
No abnormalities w ere noted.
Other findings:
Very slight or well-defined erythema was noted at the test sites of all animals. Other signs of dermal irritation noted were thickening of the skin, haemorrhage of dermal capillaries, hardened light brown or dark brown/black coloured scab, small superficial scattered scabs, scab undulating,scab cracking and scab lifting at edges to reveal bleeding, dried blood, glossy skin or further deep scabbing. Adverse reactions prevented accurate evaluation of erythema and oedema at all testsites during the study.

Table 2: Number of animals dead [and w ith evident toxicity] [and time range w ithin w hich mortality occurred]

Dose

(mg/kg bw)

Conc. In vehicle (%)*

Mortality (# dead/total)

Time range of deaths (hours)

Number with evident toxicity (#total)

Male

Female

Combined

Male

Female

Combined

2000

 

0/5

0/5

0/10

-

5/5

5/5

10/10
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Deaths did not occur in an acute dermal toxicity test in rats at 2000 mg/kg bw , therefore CNSL is of low acute dermal toxicity. Severe skin reactions were observed in all animals at the test substance application site.
Executive summary:

In an acute dermal toxicity study, groups of Sprague-Dawley rats (5/sex) were given a single dermal dose of CNSL distilled at 2000 mg/kg bw and observed for 14 days.

No mortality occurred in this limit test, therefore an LD50 has not been determined.

This acute study is classified as acceptable. It satisfies the guideline requirement for an acute dermal study in the rat.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 001 mg/kg bw
Quality of whole database:
The study is a GLP compliant and has Klimisch score 1.

Additional information

Acute oral toxicity:

Key study: The single-dose oral toxicity of the substance was performed according to the acute toxic class method OECD 423 (GLP study) in CRL:(WI) rats. Initially, three females were treated at a dose level of 2000 mg/kg bw. As no mortality was observed, a confirmatory group was treated at the same dose level. No mortality was observed in the confirmatory group, therefore no further testing was required. No test item related effects (clinical signs, body weight gain, necropsy) were observed. The acute oral LD50 value was found to be above 2000 mg/kg bw.

Acute dermal toxicity:

Key study: An acute dermal toxicity study was performed according to OECD 402 (GLP study) in Sprague-Dawley rats. Five rats per sex and per dose were treated with 2000 mg/kg bw test item under semiocclusive conditions and observed for 14 days. No mortality occurred and therefore an LD50 has not been determined (LD50 > 2000 mg/kg bw). Severe skin reactions were observed in all animals at the test substance application site (hickening of the skin, haemorrhage of dermal capillaries, hardened light brown or dark brown/black coloured scab, small superficial scattered scabs, scab undulating, scab cracking and scab lifting at edges to reveal bleeding, dried blood, glossy skin or further deep scabbing).

Discussion:

Due to data sharing obligation and following ECHA’s indications, the registrant Wave S.L. has been forced to acquire robust study summaries for acute dermal, skin and eye irritation from CNSL Consortium. This consortium didn’t allow the registrant to get access to full study reports for quality evaluation and/or proper substance identification. Consequently the present REACH registration dossier contains the available information owned by CNSL Consortium.

Wave S.L. performed an acute oral toxicity study according to OECD Guideline 423 (GLP study) with its own substance. No mortalities were observed. The substance did not cause any clinical signs during the 14 days observation period. Body weight gains during the study showed no indication of a test item-related effect, and no macroscopic observations were seen at necropsy.

On the other hand, an acute dermal toxicity study was performed by CNSL Consortium according to OECD Guideline 402 (GLP study). No deaths occurred during the study and no systemic toxicity was observed. Nevertheless, changes in the body weight were noticed alongside with local effects. Thickening of the skin, haemorrhage of dermal capillaries, hardened light brown or dark brown/black coloured scab, small superficial scattered scabs, scab undulating, scab cracking and scab lifting at edges to reveal bleeding, dried blood, glossy skin or further deep scabbing were noted. Adverse reactions prevented accurate evaluation of erythema and oedema at all test sites during the study. Although no mortality was seen, the CNSL Consortium classified the substance as Acute Toxicity Category 4, H312, based in these local effects.

The effects observed in both acute toxicity studies are inconsistent. In the acute test performed by oral route no toxicity was observed, even at necropsy. Instead, dermal local effects were observed in rats exposed by dermal route. Therefore, it can be concluded that the test item from CNSL Consortium is toxicologically different from the one of Wave S.L.

Furthermore the studies performed by CNSL Consortium are not coherent among them as well. The local dermal effects seen in the acute dermal toxicity test are however not detected in the skin irritation test. Four different CNSL samples were tested for skin irritation according to FHSA standard 16 CFR 1500.41 (GLP study). Two of the analysed samples were irritating to the skin (mean 24-72h erythema and oedema scores > 2.3) and another two were determined to be not irritating. A difference on test sample composition is a very plausible explanation.

This registrant is not allowed to perform any of these studies in vertebrates without Authorities’ permission under risk to breaching Directive 2010/63/EU of The European Parliament and of The Council of 22nd of September 2010 regarding the protection of animals used for scientific purposes which prohibit duplicate testing. However under the light of acute oral toxicity test, no acute dermal effects can be anticipated for the substance object of the present REACH Registration dossier.

Justification for selection of acute toxicity – oral endpoint

Only one study available

Justification for selection of acute toxicity – inhalation endpoint

Data waiving: Acute toxicity information provided for oral and dermal route.

Justification for selection of acute toxicity – dermal endpoint

Only one study available

Justification for classification or non-classification

Based on the results, the substance is classified for dermal acute toxicity category 4, H312 (evident toxicity observed in rats) according to CLP Regulation (EU) no. 1272/2008.