1-chloro-N,N-diethyl-1,1-diphenyl-1-(phenylmethyl)phosphoramine

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 September 2014 - 05 January 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP- and OECD TG-compliant study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000; including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

GM102E

IUPAC Name:

GM102E

Constituent 2

Reference substance name:

1-chloro-N,N-diethyl-1,1-diphenyl-1-(phenylmethyl)phosphoramine

IUPAC Name:

1-chloro-N,N-diethyl-1,1-diphenyl-1-(phenylmethyl)phosphoramine

Constituent 3

Reference substance name:

1-chloro-N,N-diethyl-1,1-diphenyl-1-(phenylmethyl)phosphoramine

EC Number:

411-370-1

EC Name:

1-chloro-N,N-diethyl-1,1-diphenyl-1-(phenylmethyl)phosphoramine

Cas Number:

82857-68-9

Molecular formula:

C23 H27 Cl N P

IUPAC Name:

benzyl(diethylamino)diphenylphosphanium chloride

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Identification: GM102E
Appearance: White powder
Molecular formula: C23H27ClNP
Molecular weight: 383.9
Batch No.: 177314
Test substance storage: At room temperature
Stable under storage conditions until: 31 August 2019 (expiry date)
Purity/composition correction factor: No correction factor required
Test substance handling: No specific handling conditions required
Stability at higher temperatures: Yes, maximum temperature: <250 °C

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: approximately 12 weeks old
- Weight at study initiation: Males: 312-374 g / Females: 210 - 235 g
- Fasting period before study: No
- Housing: Group housing of five 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
- Acclimation period: at least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24
- Humidity (%): 40 to 70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 18 September 2014 To: 24 October 2014

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

For the 0.4 mg/L exposure group, the test substance was administered to a stream of pressurized air using a combination of a brush feeder and a micronizing jet mill (Bernstein, D.N., Aerosols, pp 721- 723, 1984). A vibrator was attached to the brush feeder. The aerosol was passed through a series of three vertical cyclones, allowing larger particles to settle, and subsequently passed through the exposure chamber (Appendix 1, Figure 1). The total mean airflow was 79 L/min.

For the 0.06 mg/L exposure group, the test substance was administered to a stream of pressurized air using a combination of a spiral feeder (Randcastle Extrusion Systems, Cedar Grove, NJ, USA) and air mover (AIR-VAC, Milford, CT, USA). A vibrator was attached to the spiral feeder. The aerosol was passed through a series of three vertical and one horizontal cyclone, allowing larger particles to settle, and subsequently diluted with pressurized air before it entered the exposure chamber (Appendix 1, Figure 1). The mean total airflow was 106 L/min.

From the exposure chamber the test atmosphere was passed through a filter before it was released to the exhaust of the fume hood.

Analytical verification of test atmosphere concentrations:

yes

Remarks:

A total of 22 representative samples were taken for determination of the actual concentration during exposure at 0.4 and 0.06 mg/L, respectively.

Duration of exposure:

4 h

Concentrations:

0.06 and 0.4 mg/L

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

Mortality/Viability: Twice daily. Animals showing pain, distress or discomfort, which was considered not transient in nature or was likely to become more severe, were sacrificed for humane reasons based on OECD guidance document on humane endpoints (ENV/JM/MONO/ 2000/7). The time of death was recorded as precisely as possible.

Clinical signs - During exposure: Three times during exposure for mortality, behavioural signs of distress and effects on respiration.

Clinical signs - After exposure: On Day 1, one and three hours after exposure and once daily thereafter until Day 15. The symptoms were graded according to fixed scales and the time of onset, degree and duration were recorded: Maximum grade 4: grading slight (1) to very severe (4) Maximum grade 3: grading slight (1) to severe (3) Maximum grade 1: presence is scored (1).

Body weights: Days 1 (pre-administration), 2, 4, 8 and 15.

Necropsy: The moribund animals and animals surviving to the end of the observation period were sacrificed by an intraperitoneal injection with Euthasol® (AST Farma BV, Oudewater, The Netherlands). 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.

Statistics:

No statistical analysis was performed.

Results and discussion

Preliminary study:

Not applicable

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

>= 0.05 - < 0.5 mg/L air (nominal)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

At 0.4 mg/L, three males and two females were found dead during or within 1 hour after exposure. The remaining two males and three females were sacrificed for ethical reasons due to their poor clinical condition within 2.5 hours post-exposure.
At 0.06 mg/L, no mortality occurred.

Clinical signs:

other: At 0.4 mg/L, slow breathing, laboured respiration and excessive loss of faeces and urine was seen for the animals during exposure (not presented in the table). Lethargy, hunched posture, rales, gasping, laboured respiration, piloerection and hypothermia w

Body weight:

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

Gross pathology:

At 0.4 mg/L, macroscopic post mortem examination revealed abnormalities of the lungs (several dark red foci, dark red discolouration of the right cranial lobe, dark red colouration) for four animals sacrificed for ethical reasons. No abnormalities were seen for the remaining animals sacrificed or found dead.
At 0.06 mg/L, no abnormalities were found at macroscopic examination of the animals.

Any other information on results incl. tables

Test atmosphere characterization:

Concentration:

For the 0.4 mg/L exposure group, the time-weighted mean actual concentration was 0.42 ± 0.02 mg/L. The nominal concentration (amount of test substance used divided by the volume of pressurized air used) was 2.16 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 19%.

The concentration measurements equally distributed over time showed it was difficult to maintain a continued stable exposure level. The generation was interrupted several times in order to remove test substance deposits from the system. The generation time was elongated with 55 minutes resulting in an actual exposure time of 257 minutes. Since the actual exposure level was calculated based on the actual exposure, the achieved exposure was considered representative for one continued exposure to 0.42 mg/L for 4 hours.

For the 0.06 mg/L exposure group, the time-weighted mean actual concentration was 0.06 ± 0.002 mg/L. The nominal concentration (amount of test substance used divided by the volume of pressurized air used) was 2.64 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 2%.

The concentration measurements equally distributed over time showed it was difficult to maintain a continued stable exposure level. The generation was interrupted twice in order to remove test substance deposits from the air mover. The generation time was elongated with 8 minutes resulting in an actual exposure time of 251 minutes. Since the actual exposure level was calculated based on the actual exposure, the achieved exposure was considered representative for one continued exposure to 0.06 mg/L for 4 hours.

 

The actual concentrations deviated from the target concentrations of 0.5 and 0.05 mg/L but based on the results the interpretation and classifications were not affected.  

 

Particle size:

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice during the exposure period. At 0.4 mg/L, the MMAD was 3.1µm (gsd 1.9) and 3.6 µm (gsd 2.1). At 0.06 mg/L, the MMAD was 2.8 µm (gsd 1.5) and 3.0 µm (gsd 1.7).

Applicant's summary and conclusion

Interpretation of results:

Category 2 based on GHS criteria

Remarks:

Migrated information

Conclusions:

The inhalatory LC50, 4h value of GM102E in Wistar rats was established to be within the range of 0.05 – 0.5 mg/L.

Based on these results and according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) or Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (CLP), GM102E needs to be classified as Acute Tox. Category 2 by inhalation and labeled as H330: Fatal if inhaled.

Executive summary:

GM102E was administered as an aerosol by inhalation (nose only) for 4 hours to two groups of five male and five female Wistar rats at a concentration of 0.4 and 0.06 mg/L, respectively. Animals were subjected to daily clinical observations and determination of body weights on Days 1, 2, 4, 8 and 15.Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

For the 0.4 mg/L exposure group, the time-weighted mean actual concentration was 0.42 ± 0.02 mg/L. The nominal concentration(amount of test substance used divided by the volume of pressurized air used) was 2.16 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 19%.

For the 0.06 mg/L exposure group, the time-weighted mean actual concentration was 0.06 ± 0.002 mg/L. The nominal concentration was 2.64 mg/L resulting in a generation efficiency of 2%. The concentration measurements equally distributed over time showed that the concentrations were sufficiently stable in order to classify GM102E.  

 

The actual concentrations deviated from the target concentrations of 0.5 and 0.05 mg/L but based on the results the interpretation and classifications were not affected.  

 

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice during the exposure period.

At 0.4 mg/L, the MMAD was 3.1 µm (gsd 1.9) and 3.6 µm (gsd 2.1).

At 0.06 mg/L, the MMAD was 2.8 µm (gsd 1.5) and 3.0 µm (gsd 1.7)

 

At 0.4 mg/L, three males and two females were found dead during or within 1 hour after exposure. The remaining two males and three females were sacrificed for ethical reasons due to their poor clinical condition within 2.5 hours post-exposure.

At 0.06 mg/L, no mortality occurred.

 

At 0.4 mg/L, slow breathing, laboured respiration and excessive loss of faeces and urine was seen for the animals during exposure. Lethargy, hunched posture, rales, gasping, laboured respiration, piloerection and hypothermia were seen for the animals post-exposure on Day 1.

At 0.06 mg/L, fast breathing was seen for the animals during exposure. After exposure, hunched posture and piloerection were seen for the animals. One male showed laboured respiration. The animals had recovered from the clinical signs between Days 2 and 3.

 

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

At 0.4 mg/L, macroscopic post mortem examination revealed abnormalities of the lungs (several dark red foci, dark red discolouration of the right cranial lobe, dark red colouration) for four animals sacrificed for ethical reasons. No abnormalities seen for the remaining animals sacrificed or found dead.

At 0.06 mg/L, no abnormalities were found at macroscopic examination of the animals.

 

The inhalatory LC_{50, 4h}value of GM102E in Wistar rats was established to be within the range of 0.05 – 0.5 mg/L.

Based on these results and according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) or Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (CLP), GM102E needs to be classified as Acute Tox. Category 2 by inhalation and labeled as H330: Fatal if inhaled.