Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010-04-12 to 2010-04-26
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Sorbitan Laurate
- Physical state: yellow turbid viscous liquid
- Analytical purity: >80%
- Lot/batch No.: 0000357933
- Expiration date of the lot/batch: 27 October 2011
- Storage condition of test material: stable at RT in the dark

Note: Due to analytical test results the substance is defined as UVCB. For further details see IUCLID Chapter 1.2.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: young adult animals were selected (approximately 12 weeks old)
- Weight at study initiation: Animals used within the study were of approximately the same age and body weight variation did not exceed +/- 20% of the sex mean
- Housing: Group housing of five animals per sex per cage in labelled Macrolon cages (type IV; height 18 cm) containing sterilised sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom)
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) except during exposure to the test substance
- Water (e.g. ad libitum): Free access to tap water except during exposure to the test substance
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 (actual range: 19.8 – 21.5)
- Humidity (%): 40-70 (actual range: 34 - 53)
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: flow past nose-only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983)
- Exposure chamber volume: nose only
- Method of holding animals in test chamber: The chamber consisted of three animal sections with eight animal ports each (appendix 1, figure 1). Each animal port had its own atmosphere inlet and exhaust outlet. The animals were placed in restraining tubes and connected to the animal ports.
- Source and rate of air: at each animal port the theoretical air flow was at least 1 L/min
- System of generating particulates/aerosols: stainless steel nebulizer (type 156.000.16.16, Lechler, Metzinger, Germany)
- Method of particle size determination: The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice.
Temperature, humidity, pressure in air chamber: The actual temperature of the atmosphere was between 20.9 and 21.2°C and relative humidity was between 30.1 and 32.9%.
TEST ATMOSPHERE
- Brief description of analytical method used: Samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of the middle section of the exposure chamber. Samples were drawn through a glass fiber filter (type APFC04700, Millipore, Billerica, MA, USA). The collected amount of the non-volatile test substance in the air sample was measured gravimetrically.
- Samples taken from breathing zone: yes

TEST ATMOSPHERE (if not tabulated)
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.):
The MMAD was 4.6 and 4.7 µm, respectively and the gsd was 2.0 and 2.1, respectively

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: The target concentration was based on the EC and UN classification guidelines.

Agglomeration of aerosol particles at this high concentration might have resulted in the MMAD values to fall outside the recommended range of 1 - 4 µm. The MMAD determined (4.6 and 4.7µm) just exceeded this range. Since the gsd (2.0 and 2.1) determined during the actual exposure indicated that the aerosol was polydisperse and since approximately 40% of the particles were smaller than 4 µm,it can be assumed that sufficient deposition in the lower respiratory tract occurred during the exposure. Further testing at a lower concentration was considered but for animal welfare reasons not conducted. The exposure to the limit concentration of 5 mg/L with a slightly larger MMAD was considered of greater value than an exposure to a lower level with a MMAD closer to or in the recommended range.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
The actual concentration was determined nine times during the exposure period. Samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of the middle section of the exposure chamber.
Duration of exposure:
4 h
Concentrations:
5 mg/L
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Three times during exposure for mortality, behavioural signs of distress and effects on respiration, days 1 (pre-treatment), 2, 4, 8 and 15 for body weight changes
- Necropsy of survivors performed: yes
- Other examinations performed: All animals were sacrificed at the end of the observation period by an intraperitoneal injection with Euthasol ® (AST Farma BV, Oudewater, The Netherlands) and subsequently exsanguinated. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded. Particular attention was given to any changes in the respiratory tract.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5 000 mg/m³ air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
No mortalities occurred.
Clinical signs:
During exposure, slow breathing was observed among several animals at approximately 3 hours after start of exposure. After exposure, clinical signs recorded among all animals between Days 1 and 5 included lethargy, ventro-lateral recumbency, hunched posture, rales, shallow respiration, piloerection and/or chromodacryorrhoea. No clinical signs were observed after day 5.
Body weight:
Overall body weight gain in males and females were within the range expected for rats of this strain and age used in this type of study.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU