Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02/0215 to 04/2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): BLANCOLEN HP Disodiumhydroxy(sulfonato)acetate
- Substance type: sulfonic acid derivative
- Physical state: solid
- Analytical purity: 98 %
- Lot/batch No.: 09102303
- Expiration date of the lot/batch: 09.12.2013
- Stability under test conditions: stable
- Storage condition of test material: room temperature
Specific details on test material used for the study:
Sample Name: FA8
Batch: 14102401
Supplier: Brüggemann Chemical, L. Brüggemann KG

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Supplier: Charles River C/Argenters 7, Local AB 08290 Cerdanyola del Vallés Barcelona – Spain.
Health status: Specific Pathogen Free (SPF).
Rationale for selection of species / strain: The Sprague Dawley rat is a suitable rodent strain, acceptable to regulatory authorities as a recommended rodent test system, and for which extensive background data are available.
Total number of animals: 8: 6 used in the study (3 males and 3 females, allocated to first dose level (5.16 mg/mL) and 2 spare animals.
Age of the animals at exposure: 8 weeks
Animals per cage (before / after distribution): 4 (before distribution), 3 (after distribution)
Bedding material: Capsumlab Lecho_10 (autoclavable)
Change of cages: Once a week.
Inclusion criteria on arrival: Veterinary inspection.
Acclimatisation period: 8 days.
Animal identification: Digit ink marks.

Light cycle: 12:12, 07.00 to 19.00 CET
Pressure gradient: Animal holding room positive to corridors
Temperature: 18.6-23.5ºC
Relative humidity: 20-47%

Food: Global Diet
Reference: 2914 C
Supplier: Harlan Teklad Station Road Blackthorn, Bicester Oxon, OX25 1TP United Kingdom
Nutritional and contaminant contents:Certificate of analysis for the batch used is included in annex II
Food availability: Ad libitum, except when animals were restrained in the exposure tubes
Drinking water: Tap Water
Watering: Bottles
Quality control: Certificate of analysis is included in annex III
Water availability: Ad libitum, except when animals were restrained in the exposure tubes

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
> 1 - < 4 µm
Geometric standard deviation (GSD):
> 1.5 - < 3
Details on inhalation exposure:
Inhalation exposure was performed using a flow-past, nose-only exposure system. The animals were confined separately in restraint tubes which were positioned radially around the exposure chamber. The exposure system ensured a uniform distribution and provided a constant flow of test material to each exposure tube. The flow of air at each tube was approximately 1 L/min, which was sufficient to minimize re-breathing of the test aerosol as it is more than twice the respiratory minute volume of rats.
Exposure chambers type EC-FPC-232 (anodised aluminium, volume inside compartment: approximately 3 L), equipped with glass exposure tubes were used. The rats were individually exposed in glass tubes matching their size. Before treatment start the homogeneity for the different levels of the exposure chamber was confirmed. Cross and longitudinal sections of the chamber are shown in Figure 1.
The temperature and relative humidity of the test atmosphere in the exposure chamber was maintained as required by experimental conditions. Air flow was monitored regularly.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
gravimetric
Duration of exposure:
4 h
Concentrations:
5.16 mg/l
No. of animals per sex per dose:
3 males, 3 females
Control animals:
no
Details on study design:
The test was based on a stepwise procedure. The starting dose (5 mg/L air, during 4 hours) was selected since the test item was expected to be non-toxic according to previous studies conducted by sponsor. No additional doses were required.

3 male and 3 female rats in group A (starting dose group) were exposed for 4 consecutive hours.
During an observation period of 14 days, clinical observations and body weight were collected in order to characterise the toxicological effects of the aerosol. All animals were subjected to a gross necropsy and descriptions of all macroscopic abnormalities were recorded.

Acclimatisation to the nose-only restraining tubes was performed for approximately 80 minutes immediately before the exposure.
Statistics:
No statistical analysis was performed.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5.16 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
none
Clinical signs:
other: Dirty fur, chromorrhinorrhea, chromodacryorrhea and piloerection were observed in all animals immediately after administration and 1 hour after exposure. These clinical signs were not longer present on day 2 of study and no other clinical signs were obser
Body weight:
A slight decrease in body weight (approximately 1-2% less than body weight at pre-treatment) was observed in all animals from day 1 of study to day 2 of study. Males recovered the body weight at pre-treatment at day 3 of study. Thereafter, a normal body weight gain was observed in males. By contrast, a stagnation of body weight was observed in females, which did not recover the body weight at pre-treatment until day 6 of study. Females gained weight normally from day of study 6 onwards.
Gross pathology:
No macroscopic findings were observed during necropsy.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
No classification according to (EU) No. 1272/2008 required.
Executive summary:

Treatment of Sprague Dawley rats with the test item resulted in no deaths. Clinical signs were considered to be mainly stress related to the nose-only exposure but a contribution of the treatment with the test item cannot be completely excluded. Additionally, body weight stagnation in females is considered to be related with the test item.

It is concluded that, under the experimental conditions:

- LC50 of the test item was greater than 5.16 mg/L air (gravimetric aerosol concentration)

- Based on the GHS classification criteria, the test item can be considered as not classified.