GASIR1

Endpoint summary

Description of key information

In an OECD Test Guideline 425 study, to GLP, the acute oral LD50 of Gasir 1 was found to be greater than 2000 mg/kg bw.

In an OECD Test Guideline 403 study, to GLP, the acute inhalation LC50 of Gasir 1 was found to be greater than 5.04 mg/l.

In an OECD Test Guideline 402 study, to GLP, the acute dermal LD50 of Gasir 1 was found to be greater than 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:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)

GLP compliance:

yes

Test type:

up-and-down procedure

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 194-223g
- Fasting period before study:overnight before dosing
- Diet (e.g. ad libitum):ad libitum
- 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
- Photoperiod (hrs dark / hrs light):12hrs dar/12hrs light

Route of administration:

oral: gavage

Vehicle:

peanut oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle:no data
- Amount of vehicle (if gavage):no data
- Justification for choice of vehicle:the test material did not dissolve/suspend in distilled water

MAXIMUM DOSE VOLUME APPLIED:
The volume administered to each animal was calculated according to the fasted bodyweight at the time of dosing.

Doses:

175, 550, 2000

No. of animals per sex per dose:

175mg/kg: 1
550mg/kg: 1
2000mg/kg: 3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 30min, 1, 2 and 4 hours after dosing and subsequently once daily for 14days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:
Body weight: prior to dosing and seven and fourteen days after treatment

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Female: 2000 mg/kg bw; Number of animals: 3; Number of deaths: 0
Female: 550 mg/kg bw; Number of animals: 1; Number of deaths: 0
Female: 175 mg/kg bw; Number of animals: 1; Number of deaths: 0

Clinical signs:

other: Hunched posture was noted in the animal treated at a dose level of 550mg/kg. Signs of systemic toxicity noted in two animals treated at a dose level of 2000mg/kg were hunched posture and pilo-erection. There were nog signs of systemic toxicity noted in th

Gross pathology:

No abnormalities were noted at necropsy.

Individual bodyweights and weekly bodyweight changes

Dose level mg/kg	Animal number and sex	Bodyweight (g) at day			Bodyweight gain (g) during week
		0	7	14	1	2
175	1 -0 Female	201	239	244	38	5
550	2 -0 Female	194	209	224	15	15
2000	3 -0 Female	197	239	249	42	10
2000	4 -0 Female	223	255	260	32	5
2000	5 -0 Female	203	246	272	43	26

Interpretation of results:

GHS criteria not met

Conclusions:

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

Executive summary:

See attached documents.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 2 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

The quality of the test atmosphere fully complied with criteria documented in the respective guidelines: OECD 403, EPA OPPTS 870.1300 and Council Regulation (EC) No 440/2008.

Deviations:

yes

Remarks:

The temperature in the animal room was slightly increased (up to 0.7°C higher than the upper limit) on two days during the acclimation period. This deviation was considered to have no impact on the outcome of the study and interpretation of the results.

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:Charles River Laboratories, Research Model and Services, Germany GmbH (Sandhofer Weg 7, D-97633, Sulzfeld)
- Age at study initiation: 8-9 weeks old (sighting exposure), 9-10 weeks old (main study)
- Weight at study initiation: male: 344g, female: 214g (sighting exposure), male: 360-394g, female: 222-244g (main study)
- Housing: Group caging (5 animals, by sex, per cage), during the sighting study individual caging
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period:6 days (sighting study) or 8 days (main study)

ENVIRONMENTAL CONDITIONS
- Temperature (°C):19.5-25.7°C
- Humidity (%):40-70%
- Air changes (per hr):15
- Photoperiod (hrs dark / hrs light): 12hrs dark/12hrs light

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:TSE Rodent Exposure System (TSE Systems GmbH, Bad Homburg, Germany).
- Exposure chamber volume: 3.85L
- Method of holding animals in test chamber: polycarbonate restraint tubes
- Source and rate of air: 0.5L/min
- Method of conditioning air:
- System of generating particulates/aerosols: Rotating brush dust generator Palas RBG 1000 (Palas GmbH, Karlsruhe, Germany)
- Method of particle size determination: From the animal breathing zone using a cascade impactor
- Treatment of exhaust air: through a suitable filter system
- Temperature: 27.5°C, humidity: 9.6%, pressure in air chamber: no data

TEST ATMOSPHERE
- Brief description of analytical method used: The test atmosphere was sampled at regular intervals during the exposure period. Samples were taken from an unoccupied exposure port (representing the animal's breathing zone) by pulling a suitable volume of test atmosphere through weighed GF10 glass fibre filters (Whatman, Germany, ref. no. 10370302). The difference in the pre and post sampling weights, divided by the volume of atmosphere sampled, was equal to the actual achieved test atmosphere concentration.
- Samples taken from breathing zone: yes. Filter samples were collected at the breathing zone (approx. every 10-20 minutes) during each 4-hour exposure period and analyzed.

VEHICLE
- Composition of vehicle (if applicable):
- Concentration of test material in vehicle (if applicable):
- Justification of choice of vehicle:
- Lot/batch no. (if required):
- Purity:

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: The particle size of the test atmosphere was determined three times during the exposure period using a 7-stage impactor of Mercer style (TSE systems GmbH, Bad Homburg, Germany).
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): 2.53µm (MMAD), 2.38 (GSD) (=sighting group); 2.41µm (MMAD), 2.32 (GSD) (=main study).

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

5.04mg/l

No. of animals per sex per dose:

1 male/female (sighting exposure)
5 male/female (main study)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Morbidity/mortality: 1 hour after exposure and twice daily during the 14-day observation period. Clinical signs: hourly intervals during exposure and twice a day the day of exposure and once daily for 14 days. Bodyweight: on day 1, 3, 7, 14.
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.04 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No animals died during the study.

Clinical signs:

other: In group 0.1 (sighting exposure): slightly to moderate laboured respiration was recorded up to 1 hour after exposure. In group 1 (main study): slight laboured respiration was observed in all animals during the exposure and shortly thereafter when removed

Body weight:

In Group 0.1 as sighting group, slight body weight loss was observed in the male animal. The body weight returned to the initial values on Day 3.
In Group 1, slight body weight loss (1-8%) was observed in all animals, except two female rats (no.: 8236, 8239) where moderate body weight loss (11-12%) was recorded. The body weight of 4 males (no.: 8207, 8210, 8211, 8212) and one female (no.: 8242) returned to the initial values on approximately on Day 3, while a single male (no.: 8209) and 4 female animals (no.: 8236, 8239, 8240, 8241) gained their initial body weight back on approximately Day 7.

Gross pathology:

There was no evidence of any macroscopic observations at a concentration of 5.04 mg/l in animals terminated on day 14.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions of this study, no lethality occurred in Group 0.1 or Group 1 when exposed to an aerosol concentration of 5.04 mg/L each for 4 hours. The acute inhalation median lethal concentration (LC50) of GASIR1 in Wistar Crl:WI rats was therefore considered to be above 5.04 mg/L.

Executive summary:

1.1. STUDY DESIGN

Wistar Crl:WI rats were exposed to a test atmosphere of GASIR1 at concentration of 5.04 mg/L. The test item was administered as supplied by the Sponsor without any preparation.

The study was performed in two steps. A sighting exposure was performed first, where test atmospheres at concentrations of 5.04 mg/L was tested on single animals of both sexes. No lethality was observed at this concentration. Thereafter the main study was performed at a concentration of 5.04 mg/L. Five male and five female rats were used in the main group (Group 1). In both study phases, the animals were exposed to the test atmosphere for 4 hours using a nose-only exposure system. Aerosol concentration was measured gravimetrically 18 times during each 4-hour exposure in both parts of the study and the particle size distribution of the test aerosol was determined 3 times. The day of exposure was designated Day 0 in both parts of the study and a 14-day observation period followed the exposures.

Clinical observations were performed for all animals during exposure at hourly intervals, following removal from restraint, approximately 1hour following the end of the exposure, and daily for 14 days thereafter. Body weight was measured on Days 0 (before the exposure), 1, 3, 7 and 14. Gross necropsy was performed on the animal that died and on all animals sacrificed on Day 14.

No control group was exposed in this study.

1.2. RESULTS

The quality of the test atmosphere fully complied with criteria documented in the respective guidelines: OECD 403, EPA OPPTS 870.1300 and Council Regulation (EC) No 440/2008.

The mean achieved atmosphere concentration in the study was 5.04 mg/L in both Group 0.1 and Group 1. The mass median aerodynamic diameter (MMAD) was 2.53 μm and 2.41μm with geometric standard deviation (GSD) 2.38 and 2.32 in the Sighting Group and the Main Group, respectively.

Sighting Exposure

Initially, a single male and female rats (Group 0.1) were exposed to the aerosol concentration of 5.04 mg/L. Both animals survived.

The main clinical sign was laboured respiration, slight in the male rat and slight to moderate in the female animal. A slight body weight loss was also observed on Days 0-3 in the male of Group 0.1. The body weight returned to the initial value on Day 3.

At the necropsy, no external or internal finding was recorded in Group 0.1.

Main Study

Mortality

No mortality was noted in Group 1, either during the exposure or during the 14 day observation period.

Clinical findings

In Group 1, slight laboured respiration was observed in all animals during the exposure and shortly thereafter when removed from the restraint tubes. Increased respiratory rate (slight) was also recorded in 3 out of 5 males and in 2 out of 5 females on Day 0. Slightly weak body condition was noted in 2/5 males and 4/5 females and persisted for up to Day 2 in males and up to Day 6 in females. In a single female, moderately weak body condition was observed on Day 1 and Day 2. From Day 7 all animals were symptom-free.

Additionally, fur stained by test item was commonly observed in all animals following the exposure and persisting for up to Day 5. Wet fur was also generally recorded in all animals on the day of exposure. This observation was considered to be related to the restraint and exposure procedures and was considered not to be toxicologically significant.

Body weight

In Group 1, slight body weight loss (1-8%) was observed in all animals, except two female rats where moderate body weight loss (11-12%) was recorded. The body weight of 4 males and one female returned to the initial values on approximately on Day 3, while a single male and 4 female animals gained their initial body weight back on approximately Day 7.

Necropsy

No external or internal finding was recorded at necropsy in Group 1.

1.3. CONCLUSION

Under the experimental conditions of this study, no lethality occurred in Group 0.1 or Group 1 when exposed to an aerosol concentration of 5.04 mg/L each for 4 hours. The acute inhalation median lethal concentration (LC50) of GASIR1 in Wistar Crl:WI rats was therefore considered to be above 5.04 mg/L.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

> 5.04 mg/L air

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:

2011-07-20 to 2011-08-03

Reliability:

1 (reliable without restriction)

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:Laboratoire Elevage Janvier, B.P. 4105, Route des Chênes Secs, 53940 Le Genest-St-Isle CEDEX France
- Age at study initiation: Young adult rats
- Weight at study initiation: between 210 and 272g
- Housing: individual caging
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.1-24 °C
- Humidity (%): 42-69%
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12hrs dark/12hrs light

Type of coverage:

semiocclusive

Vehicle:

other: none

Details on dermal exposure:

TEST SITE
- Area of exposure:10% of the total body surface
- % coverage: 100%
- Type of wrap if used: semi occlusive plastic wrap

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washed after exposure (24h) with water of body temperature
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000mg/kg bw
- Constant volume or concentration used: yes
- For solids, paste formed: no, the test item was placed onto a gauze pad and moistened with water

VEHICLE
- Amount(s) applied (volume or weight with unit): one gauze pad
- Lot/batch no. (if required): 102/26

Duration of exposure:

24h hours

Doses:

2000mg/kg bw

No. of animals per sex per dose:

5 animals/sex

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Clinical observation: 1, 5 hours after treatment, then daily for 14 days. Body weighing: day 0, 7, 14
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

approximate LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occured after a 24 hour dermal exposure to GASIR 1 followed by a 14-day observation period

Clinical signs:

other: No clinical signs were observed

Other findings:

Small testes or epididymides, pelvic dilatation of the left kidney and luminal dilatation of the uterine horns were incidentally seen at necropsy.
Local dermal signs: Russet staining was recorded on the skin in all animals after dosing. The discoloration of the skin lasted up to day 4 in the male and day 8 in the female animals. No other local dermal signs were observed.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute dermal median lethal dose (LD50) of the test item GASIR was found to be higher than 2000mg/kg body weight in male and female RjHan:(WI) Wistar rats.

Executive summary:

An acute dermal toxicity study was performed with test item GASIR1 in RjHan:(WI) Wistar rats, in compliance with OECD Guideline No.: 402.

A limit test was carried out at 2000mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as supplied, moistened with water, as a single dermal 24-hour exposure followed by a 14-day observation period.

Clinical observations were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. Rats were euthanized and a gross macroscopic examination performed at the end of the 2-week observation period (Day 14).

The results of the study were summarized as follows:

Mortality

No mortality occurred.

Systemic clinical signs

No clinical signs were observed after the treatment with the test item or during the 14-day observation period.

Local dermal signs

No local dermal signs were observed during the entire study period. However, russet staining was observed on the skin in all animals after dosing from Day 1 to Day 4 in the males and Day 1 to Day 8 in the female animals.

Body weight

The body weight and body weight gain of GASIR1 treated animals did not show any test item-related effect.

Necropsy

There was no evidence of the test item-related observations at a dose level of 2000 mg/kg bw at necropsy. Small testes or epididymides, pelvic dilatation of the left kidney and luminal dilatation of the uterine horns were incidentally seen.

Conclusions

The acute dermal median lethal dose (LD50) of the test item GASIR1 was found to be higher than 2000 mg/kg bw in male and female RjHan:(WI) Wistar rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 2 000 mg/kg bw

Justification for classification or non-classification

Based on the results of the available and reliable acute oral, dermal and inhalation studies, no classification is required for Gasir 1 according to EU CLP.