Registration Dossier

Administrative data

Description of key information

Rat oral LD50 > 5000 mg/kg bw

Rat ihalatory LC50 > 1895 mg/m3

Rat dermal LD50 > 2000 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:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted according to internationally accepted testing guidelines and performed according to GLP. Nevertheless the test substance purity is low. Justification for Read Across is reported in the endpoint summary and in the Category Justification Report attached to Section 13 of this dossier.
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd., Manston, Kent, U.K.
- Age at study initiation: approximately five to eight weeks old.
- Weight at study initiation: at the start of the main study the males weighed 122 - 140 g, and the females 120 - 126 g.
- Fasting period before study: not access to drink and food an overnight fast immediately before dosing and for approximately two hours after dosing.
- Housing: animals were housed in groups of up to five by sex in solid-floor polypropylene cages with sawdust bedding.
- Diet: free access to food (Rat and Mouse Expanded Diet No. 1, Special Diet Services Limited, Witham, Essex, U.K.).
- Water: free access to mains drinking water.
- Acclimation period: minimum acclimatisation period of at least five days.

ENVIRONMENTAL CONDITIONS
- Temperature: 19-21°C
- Humidity: 43-55%
- Air changes: 15 changes per hour.
- Photoperiod: lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
All animals were dosed once only by gavage using a metal cannula attached to a graduated syringe.
The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
Group 1: 1 male and 1 female (observation period of 5 days)
Group 2: 5 males and 5 females (observation period of 14 days).
Details on study design:
- Duration of observation period following administration: a first group (group 1) was observed for deaths and over signs of toxicity for 5 days; the second group (group 2) was observed for deaths and over signs of toxicity for 14 days.
- Frequency of observations and weighing: individual bodyweights were recorded on the day of dosing in the group 1, prior to dosing on day 0 and on days 7 and 14, or at death in group 2.
- Necropsy of survivors performed: no in the group 1; in the group 2 at the end of the study the surviving animals were killed by cervical dislocation.
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: all animals in the group 2 were subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Group 1: there were no deaths.
Group 2: one male was killed one hour after dosing having sustained a broken leg. This was done for humane reasons in accordance with current U.K. Home Office guidelines. The death was therefore not attributed to the toxicity of the test material.
Clinical signs:
Group 1: there were no clinical signs of toxicity.
Group 2: no signs of systemic toxicity were noted during the study.
Body weight:
Group 2: surviving animals showed expected gain in bodyweight during the study.
Gross pathology:
Group 2: no abnormalities were noted at necropsy.

Individual clinical observations and mortality data

Dose level (mg/kg bw) Group Animal number & sex Effects noted after dosing (hours) Effects noted after dosing (hours)
1/2 1 2 3 4 1 2 3 4 5 6 7 8 9 10 11 12 13 14
2000 1 1-0 M 0 0 0 0 0 0 0 0 0 0 - - - - - - - - -
2-0 F 0 0 0 0 0 0 0 0 0 0 - - - - - - - - -
2000 2 3-0 M 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
3-1 M 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
3-2 M 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
3-3 M 0 0X - - - - - - - - - - - - - - - - -
3-4 M 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
4-0 F 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
4-1 F 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
4-2 F 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
4-3 F 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
4-4 F 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

0: no signs of systemic toxicity

X: animal killed due to broken leg

Interpretation of results:
not classified
Remarks:
Migrated information According to the CLP regulation. Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg body-weight.
Executive summary:

Method

A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines No. 401 "Acute Oral Toxicity" referenced as Method B1 in Commission Directive 84/449/EEC.

Following a range-finding study (group 1), a group of ten fasted animals (group 2) was given a single oral dose of undiluted test material, at a dose level of 2000 mg/kg bodyweight. The surviving animals were observed for fourteen days after the day of dosing and were then killed for gross pathological examination.

One male was killed one hour after dosing having sustained a broken leg. This was done for humane reasons in accordance with current U.K. Home Office guidelines. The death was therefore not attributed to the toxicity of the test material.

Results

No signs of systemic toxicity were noted during the study. Surviving animals showed expected gain in bodyweight during the study. No abnormalities were noted at necropsy.

The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg body-weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Particle size not specified. The analogous substance, part of the Stilbene Fluorescent Withening Agents, group 3, is the acid form of the disulphonated derivative dihydroxyethyl derivative, therefore contains the same organic functional groups, but due to the sulphonation/salification degree it is less soluble. This property makes of Read Across substance 4404-43-7 a conservative representative because of the potential higher bioavailability. Justification for Read Across is reported in the endpoint summary and in the Category Justification Report attached to Section 13 of this dossier.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
GLP compliance:
no
Remarks:
Pre GLP.
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Borchen, Germany.
- Weight at study initiation: 180 - 200 g.
Route of administration:
inhalation: dust
Type of inhalation exposure:
not specified
Vehicle:
air
Analytical verification of test atmosphere concentrations:
yes
Remarks:
gravimetric with membrane filter.
Duration of exposure:
1 - 4 h
Concentrations:
163.3, 375, 1225 and 1895 mg/m³ air at 4 hour exposure.
1820 mg/m³ air at 1 hour exposure.
No. of animals per sex per dose:
10
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.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 1 895 mg/m³ air (nominal)
Based on:
test mat.
Exp. duration:
4 h
Sex:
male/female
Dose descriptor:
LC50
Effect level:
1 820 mg/m³ air (nominal)
Based on:
test mat.
Exp. duration:
1 h
Mortality:
No mortality occuerred.
Clinical signs:
other: At concentrations of 1225 and 1895 mg/m³ air at the 4 hour exposure the animals showed a decreased general condition for about 4 - 6 hours.
Gross pathology:
No abnormalities detected.
Interpretation of results:
other: not applicable.
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
LC50 (4 h): > 1.895 mg/l air
Executive summary:

Method

Acute inhalation toxicity was assessed following the method described into the OECD guideline 403.

Results

LC50 (4 h): > 1.895 mg/l air

LC50 (1 h): 1.820 mg/l air

Conclusion

According to CLP Regulation (EC 1272/2008), 3.1 Acute toxicity section, substances can be allocated to one of four toxicity categories based on acute toxicity by the oral, dermal or inhalation route according to the numeric criteria. Acute toxicity values are expressed as (approximate) LD50 (oral, dermal) or LC50 (inhalation) values or as acute toxicity estimates (ATE).

The inhalation LD50 value classification limit for dust is 5.0 mg/l (category 4: 1.0 < ATE ≤ 5.0 mg/l). In the current test an LD50 was non identified; considering the fact that no mortality occurred, a classification category can not be assigned. Thus, a classification according to the CLP Regulation (EC 1272/2008) is not applicable.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Value:
1 895 mg/m³

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted according to internationally accepted testing guidelines and performed according to GLP. Justification for Read Across is reported in the endpoint summary and in the Category Justification Report attached to Section 13 of this dossier.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wblferstrasse 4, CH-4414 Fullinsdorf.
- Age at study initiation: 10 weeks (males), 12 weeks (females).
- Weight at study initiation: 225 - 247 g (males), 200 - 222 g (females).
- Housing: individually in Makrolon type-2 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz).
- Diet: pelleted standard Kliba 343, Batches 77/90 and 78/90 rat maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst); ad libitum.
- Water: community tap water from Itingen; ad libitum.
- Acclimation period: one week.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 40-70 %
- Air changes: 10 - 15 air changes per hr.
- Photoperiod: 12 / 12 hrs dark / hrs light.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: backs of the animals.
- Preparation: clipping 24 h before application.
- % coverage: 10% of the total body surface.

REMOVAL OF TEST SUBSTANCE
- Washing: with lukewarm tap water.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount applied: 4 ml
- Concentration: 100%
- For solids, paste formed: yes
Duration of exposure:
24 h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: observations four times during test day 1, and daily during days 2 - 15; weighing on test days 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: gross pathology.
Statistics:
The LOGIT-Model could not be applied to the observed rate of death. The toxicity was estimated without use of a statistical model.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality.
Clinical signs:
No systemic signs were observed in the animals during the entire observation period.
Skin observations: males/females: scales (back) (10/10); general erythema (back) (3/10); males: erythema focal (back) (3/5). All animals had recovered from the local signs after 7 observation days.
Body weight:
Mean body weight (g): days 1, 8, 15
- males: 236.2, 265.2, 294.8
- females: 207.3, 214.0, 217.8
Gross pathology:
No findings noted.
Interpretation of results:
not classified
Remarks:
Migrated information According to he CLP Regulation. Criteria used for interpretation of results: EU
Conclusions:
LD50 > 2000 mg/kg bw
Executive summary:

Method

Upon an acute oral single administration (2000 mg/kg bw) and a 15 day post-treatment observation period, the dermal LD50 was determined for the test substance, according to the OECD guideline 402.

Results

LD50 > 2000 mg/kg bw

No deaths accorred and no systemic signs were observed in the animals during the entire observation period. Skin reactions are all recovered after 7 observation days.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

No specific tests on the registering substance have been performed, but the substance is part of the Stilbene Fluorescent Whitening Agent category, goup 3, in which all members share the common organic functional group dihydroxyethylamino derivative, with different sulphonation degrees: acid form (CAS 4404-43-7), disulphonated sodium salt (CAS 4193-55-9), disulphonated sodium/potassium form (CAS 70942-01-7) tetrasulphonated (CAS 16470-24-9) and hexa sulphonated (CAS 68971 49-3).

For three representative substances valid tests have been performed according to OECD guidelines which resulted in no effect up to the highest tested dose (5000 mg/Kg bw). The result for the registering substance can be well interpolated between results for the other substances: the less soluble CAS 4193-55-9 /CAS 70942-01-7 and the more soluble CAS 16470-24-9.

Within the whole category ten over fourteen substances have been tested and none of the existing tests arised any concern for acute oral toxicity

Skin adsorption has been evaluated and calculated for all members of the category (See Category Justification Report, Section 13)

As it can be noted, the influence of the variability in functional group is very low, more related to the variability in the polarity of the substance than on potential reactivity that can arise a concern. From a metabolic point of view an estimation with OECD Toolbox of the dermal metabolism has been performed in order to verify if breakdown products could be formed. in the category 7 members have been tested for acute oral toxicity, representing several subgroups and results indicated a LD50 > 2000 (the highest tested dose) for all members.

A good GLP Klimish 1 study for acute dermal toxicity has been reported for CAS 16470-24-9 (Ciba-Geigy Ltd., 1991), performed at 2000 mg/Kg bw with no effect.

Because of the physical state and the trade forms, inhalation is not an appropriate route of exposure. Acute toxicity results for the other two exposure routes indicate no concern therefore no testing has been performed. The result of a test performed on an analogous substance at the maximum allowed concentration of 1890 mg/m3has been reported, where no effect is indicated. The analogous substance, part of the Stilbene Fluorescent Withening Agents, group 3, is the acid form of the disulphonated derivative dihydroxyethyl derivative, therefore contains the same organic functional groups, but due to the sulphonation/salification degree is less soluble. This property makes of Read Across substance 4404-43-7 a conservative representative because of the potential higher bioavailability

Because of the physical state and the trade forms, inhalation is not an appropriate route of exposure. Acute toxicity results for the other two exposure routes indicate no concern therefore no testing has been performed. The result of a test performed on an analogous substance at the maximum allowed concentration of 1890 mg/m3has been reported, where no effect is indicated.

As a conclusion it can be stated that the substance is not acutely toxic for all the three exposure ways

Justification for selection of acute toxicity – oral endpoint

Study conducted according to internationally accepted testing guidelines

Justification for selection of acute toxicity – dermal endpoint

Study conducted according to internationally accepted testing guidelines, in GLP. Furthermore the active ingredient tested was 83%.

Justification for classification or non-classification

According to the CLP Regulation (EC1272/2008), 3.1 Acute toxicity section, substances can be allocated to one of four toxicity categories based on acute toxicity by the oral, dermal or inhalation route according to the numeric criteria. Acute toxicity values are expressed as (approximate) LD50 (oral, dermal) or LC50 (inhalation) values or as acute toxicity estimates (ATE).

The oral LD50 value was established to be greater than 5000 mg/kg body weight, therefore the test substance is out of any classification limit for acute oral toxicity (oral acute toxicity category 4: 300 < ATE ≤ 2000 mg/kg bw).

The dermal LD50 value was established to exceed 2000 mg/kg body weight, which exceeded the highest CLP classification limit (dermal acute toxicity category 4: 1000 < ATE ≤ 2000 mg/kg bw).

In conclusion, the available experimental data is adequate for classification and labelling and

the test substance is non classified for oral/dermal acute toxicity, according to the CLP Regulation (EC 1272/2008).