D-Glucitol, 1-deoxy-1-(methylamino)-, N-C8-10 acyl derivs.

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 January - 11 February 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C8-10(even numbered) acyl] derivs.

EC Number:

940-284-1

Cas Number:

1591782-62-5

Molecular formula:

C15H31NO6 C17H35NO6

IUPAC Name:

D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C8-10(even numbered) acyl] derivs.

Constituent 2

Reference substance name:

Synergen GA (50% Glucamide 810 in water)

IUPAC Name:

Synergen GA (50% Glucamide 810 in water)

Test material form:

other: liquid

Details on test material:

- Substance type: Clear slightly yellow liquid
- Physical state: liquid
- Purity: 50% active in water
- Storage condition of test material: In refrigerator (2-8°C) in the dark
- Purity/composition correction factor required: No
- General information: Liquid is self-preserving. If diluted with water, the substance is stable for 24 hours at room temperature, after that, the sample should not be used anymore as microbial growth may happen.
- Hygroscopic: No
- Volatile: No
- Specific Gravity / Density: 1.08
- pH: 8-10 at concentration of 1% in water
- Stability at higher temperatures: Yes, maximum temperature: 50°C, maximum duration: 72 hours
- Stability in Water: At least for 24 hours
- Solubility in Water: Miscible

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals (approximately 12 weeks old) were selected.
- Weight at study initiation: Body weight variation did not exceed +/- 20% of the sex mean.
- Housing: Group housing of three animals per sex per cage in labelled Makrolon cages (type IV; height 18 cm) containing sterilised sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) 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.
- Acclimatization period: at least 5 days before start of treatment under laboratory conditions.
- Health inspection: A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health.

ENVIRONMENTAL CONDITIONS
Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, at least 10 air changes/hour and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.

IN-LIFE DATES: 23 January - 11 February 2014

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

other: Water, the test substance could not be aerosolized undiluted.

Details on inhalation exposure:

The design of the exposure chamber is based on the flow past nose-only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983). The chamber consisted of three animal sections with eight animal ports each. Each animal port had its own atmosphere inlet and exhaust outlet. The animals were placed in restraining tubes and connected to the animal ports. The number of animal sections and number of open inlets were adapted to the air flow in such a way that at each animal port the theoretical air flow was at least 1 L/min, which ensures an adequate oxygen supply to the animals. The main inlet of the test atmosphere was located at the top section and the main outlet was located at the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal.
All components of the exposure chamber in contact with the test material were made of stainless steel, glass, rubber or plastic. To avoid exposure of the personnel and contamination of the laboratory the exposure chamber was placed in a fume hood, which maintained at a slight negative pressure.

Analytical verification of test atmosphere concentrations:

no

Duration of exposure:

4 h

Concentrations:

5.1 and 1.1 mg/L

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

The test substance was mixed with water (1:1 v/v). No correction was made for the specific gravity or the purity of the test substance. The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. The formulations were administered as an aerosol by inhalation for 4 hours to two groups of three male and three female Wistar rats each. Animals were subjected to daily observations and determination of body weights on Days 1, 2, 4, 8 and 15 and at death. Macroscopic examination was performed on the day of death or after terminal sacrifice (day 15).

For the 5 mg/L exposure group, the mean actual concentration was 5.1 ± 0.13 mg/L. The nominal concentration was 9.1 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 56%. The concentration measurements equally distributed over time showed that the substance was sufficiently stable. The variations were caused by adjustments to the generation equipment and were considered not to have affected the exposure level. The generation was interrupted twice during one minute in order to clean the nebulizer. The generation time was elongated with 2 minutes in order to achieve an actual exposure time of 240 minutes.

For the 1 mg/L exposure group, the mean actual concentration was 1.1 ± 0.03 mg/L. The nominal concentration was 1.9 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 56%. The concentration measurements equally distributed over time showed that the substance was sufficiently stable. The variations were caused by adjustments to the generation equipment and were considered not to have affected the exposure level. The generation was interrupted for 2 minutes in order to clean the nebulizer. The generation time was elongated with 2 minutes in order to achieve an actual exposure time of 240 minutes.

Statistics:

No statistical analysis was performed.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

other: LC50 cut-off

Effect level:

5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

At 5 mg/L, two males and one female were found dead within 3 days post-treatment. No further mortality occurred.

Clinical signs:

other: At 5 mg/L, the animals showed slow breathing and one animal was restless during the exposure. After exposure, lethargy, hunched posture, abnormal gait, laboured respiration, rales, chromodacryorrhoea and/or ptosis was noted among the animals between Days

Body weight:

Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity.

Gross pathology:

Macroscopic post mortem examination of the males found dead revealed several dark red foci on the longs or foamy contents in the lungs. Macroscopic post mortem examination of the female found dead and of the surviving animals at termination did not reveal any abnormalities. Incidental findings included advanced autolysis which was considered not related to treatment.

Applicant's summary and conclusion

Interpretation of results:

harmful

Remarks:

Migrated information category 4 Criteria used for interpretation of results: OECD GHS

Conclusions:

The inhalatory LC50 cut-off, 4h value of Synergen GA (50% Glucamide 810 in water) in Wistar rats was established to be 5 mg/L.
According to the OECD 436 test guideline, the LC50, 4h cut-off value was considered to be 5 mg/L. Based on these results and according to the:
- Globally Harmonized System of Classification and Labelling of Chemicals (GHS), United Nations, New York and Geneva (2011) (including all amendments),
Synergen GA (50% Glucamide 810 in water) should be classified in Class 4 for acute toxicity by the inhalation route.
- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments),
Synergen GA (50% Glucamide 810 in water) should be classified as Category 4 and should be labeled as H332: Harmful if inhaled.

Executive summary:

The assessment of acute inhalation toxicity of a 50% solution (in water) of Glucamide 810 (Test material Synergen GA) was performed using the acute toxic class method according to OECD TG 436. For technical reasons a 50% formulation of Glucamide 810 in water was used for the experiment. The test material was administered as aerosol by inhalation for 4 hours to two groups of 3 male and 3 female Wistar rats at analytical verified exposure concentration of 1.1 mg/L and 5.1 mg/L per group. The subsequent observation period was 14 days (terminal sacrifice on Day 15). At 5 mg/L, unspecific clinical signs of toxicity consisting of lethargy, hunched posture, abnormal gait, laboured respiration, chromodacryorrhoea and/or ptosis were observed between Days 1 and 9. Two male and one female animal were found dead within 3 days post-treatment. No further mortality occurred. At 1 mg/L neither clinical signs nor mortality was noted. The inhalatory LC₅₀ cut-off value of a 50% Glucamide 810 in water in Wistar rats was 5 mg/L.  As such, it was concluded to be category 4 via the inhalation route on the GHS scale.