Reaction mass of bis[4-(vinyloxy)butyl] hexane-1,6-diylbiscarbamate and 9,18-dioxo-3,8,19-trioxa-10,17-diazapentacos-1-en-25-yl [6-({[4-(vinyloxy)butoxy]carbonyl}amino)hexyl]carbamate

Endpoint summary

Description of key information

Acute studies are available with exposure via the oral, dermal and inhalation route. All studies were performed according to OECD/ EC guidelines and under GLP principles. None of the studies show acute toxicity.

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

Study period:

02 July, 2013 - 18 July, 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

(2001)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

(2008)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

(2002)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No 8147, April 2011, including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

other: Crl:WI (Han)

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals (approx. 8 weeks old)
- Weight at study initiation: 142 - 169 g
- Fasting period before study: overnight prior to dosing and until 3-4 hours after administration of the test substance.
- Housing: Group housing of 3 animals per cage in labeled Makrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): approx 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

GAVAGE METHOD: plastic feeding tubes.

Frequency: single dosage, on Day 1.

VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg body weight.

DOSAGE PREPARATION: The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test substance.

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

6 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily.
Body weights: Days 1 (pre-administration), 8 and 15.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy.
- Other examinations performed: none.

Statistics:

No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Hunched posture was seen only on the day of treatment for three of the six animals.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

not classified

Conclusions:

In an acute oral toxicity study with rats, performed according to OECD 423 test guideline and GLP principles, an LD50 >2000 mg/kg bw was determined.

Executive summary:

The test substance was assessed for toxicity in an acute oral toxicity study with female Wistar rats, performed according to OECD 423 test guideline and GLP principles. No mortality occurred. Hunched posture was seen only on the day of treatment for three of the six animals. No abnormalities were found at macroscopic post mortem examination of the animals. Based on the results, the oral LD50 value of URACROSS ZW7672P, Product ID 021116/000 in Wistar rats was established to exceed 2000 mg/kg body weight.

According to Regulation (EC) No 1272/2008, URACROSS ZW7672P, Product ID 021116/000 does not have to be classified for acute oral toxicity.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

A reliable study is available (Klimisch 1 study).

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

Study period:

03 October 2016 - 17 October 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The study was performed as a data requirement for a registration in China.

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

September 2009

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

May 2008, including most recent amendments

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Version / remarks:

August 1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147

Version / remarks:

November 2000; including the most recent partial revisions

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

d.d. 03 November 2015

Test type:

fixed concentration procedure

Limit test:

yes

Species:

rat

Strain:

other: Crl: WI(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 10-11 weeks old
- Weight at study initiation: males: 293-314 g; females: 193-211 g
- Fasting period before study: no
- Housing: Group housing of 5 animals in Makrolon cages (type IV; height 18 cm) containing sterilised sawdust as bedding material and paper as cage-enrichment
- Diet: pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany), ad libitum except during exposure to the test item
- Water: tap water ad libitum except during exposure to the test item
- Acclimation period: 5 days

Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed according to facility standard procedures. There were no findings that could interfere with the study.

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

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

> 3.8 - < 5.2 µm

Geometric standard deviation (GSD):

1.8

Remark on MMAD/GSD:

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice during the exposure period. The MMAD was 5.2 μm (gsd 1.8) and 3.8 μm (gsd 1.8). The first measurement just felt outside the recommended range of 1 - 4 μm. There was no evidence for test item deposition in the upper airways due to this larger MMAD. Since the second measurement felt within the recommended range and since it is generally known that good distribution throughout the lung requires particles with an aerodynamic diameter between 1 and 5 μm, it can be assumed that sufficient deposition in the lower respiratory tract occurred during the exposure.

Details on inhalation exposure:

TEST ITEM PREPARATION:
Before use the test item was grinded with an automatic grinder (Retsch PM100 Planetary Ball Mill, Retsch GmbH, Haan, Germany) and additionally with a small automatic grinder. The test item was passed through a 250 μm steel mesh sieve before use.

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: exposure chamber based on the directed flow nose only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983)
- Exposure chamber volume: ca. 150 mL
- Test chamber: polycarbonate restraining tubes
- Source and rate of air: at least 1 L/min (theoretical air flow in each animal port).
- System of generating aerosols: The test item was fed to a stream of pressurized air (mean air flow 11 L/min) by means of a spiral feeder (Randcastle Extrusion Systems, Cedar Grove, NJ, USA) and a micronizing jet mill (Bernstein, D.N., Aerosols, pp 721- 723, 1984). The aerosol was passed through a series of three cyclones, allowing larger particles to settle, before it entered the exposure chamber.
From the exposure chamber the test atmosphere was passed through a filter before it was released to the exhaust of the fume hood.
- Method of particle size determination: The particle size distribution was characterized twice during the exposure period. The samples were drawn (2 L/min) from the test atmosphere through a tube mounted in one of the free animal ports. The samples were collected with an 8 stage Marple personal cascade impactor containing fiber glass filters and a fiber glass back-up filter. Amounts of test item collected were measured gravimetrically. Subsequently the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation (GSD) were determined.
- System stability: An indication of the stability of the test atmosphere was obtained from the concentration measurements, which were equally distributed over time.
- Temperature and humidity in air chamber: 21.1-21.6 °C and 54-60% respectively

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. The collected amount of the test item in the air sample was measured gravimetrically. Sample volumes were measured by means of a dry gas meter. The time-weighted mean concentration with the standard deviation was calculated.
- Samples taken from breathing zone: Yes, the actual concentration was determined twenty-one times during the exposure period.
- The temperature and relative humidity were measured with a humidity and temperature indicator (E+E Elektronik, Engerwitzdorf, Austria) and were recorded after the animals were placed in the experimental set-up and at 30 minute intervals after initiation of the exposure.
CLASS METHOD
- Rationale for the selection of the starting concentration: Target concentrations were based on the cut off concentration values specified in the UN and EC classification guidelines.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

5 mg/L

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:
- During exposure: 3 times
- After exposure:
Mortality: Twice daily
Clinical signs: On day 1, one and three hours after exposure and once daily thereafter until day 15
Body weights (individually): on day 1(pre-administration), 2, 4, 8 and 15.

- Necropsy of survivors performed: yes

Statistics:

No statistical analysis was performed.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 4.7 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortality occurred.

Clinical signs:

other: One male showed slow breathing during exposure. One male showed piloerection one hour after exposure. No further clinical signs were noted.

Body weight:

No abnormalities observed. 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

Gross pathology:

No abnormalities were found at necropsy

Other findings:

- Actual exposure concentrations: The time-weighted mean actual concentration was 4.7 ± 0.13 mg/L. The nominal concentration (amount of test item used divided by the volume of pressurized air used) was 513 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 0.9%.

- Particle size: The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice during the exposure period.(see table 1)

Table 1 Aerodynamic particle size distribution in the test atmosphere

Measurement 1:

Stage	Cut point (mm)		Mass sampled (mg)	Relative mass (%)	Cumulative mass (% of total sampled)
1	21.0		0.06	0.93	99.07
2	15.0		0.14	2.17	96.90
3	10.0		1.01	15.66	81.24
4	6.0		0.91	14.11	67.13
5	3.5		3.00	46.51	20.62
6	2.0		1.11	17.21	3.41
7	0.9		0.22	3.41	0.00
8	0.5		0.00	0.00	0.00
Back up	0.25		0.00	0.00	0.00
MMAD^1(μm):		5.2
gsd^2:		1.8

Measurement 2:

Stage	Cut point (mm)		Mass sampled (mg)	Relative mass (%)	Cumulative mass (% of total sampled)
1	21.0		0.05	0.63	99.37
2	15.0		0.00	0.00	99.37
3	10.0		0.42	5.26	94.11
4	6.0		2.08	26.07	68.05
5	3.5		3.56	44.61	23.43
6	2.0		1.20	15.04	8.40
7	0.9		0.36	4.51	3.88
8	0.5		0.12	1.50	2.38
Back up	0.25		0.19	2.38	0.00
MMAD^1(μm):		3.8
gsd^2:		1.8

¹ Mass Median Aerodynamic Diameter;² Geometric standard deviation

Interpretation of results:

GHS criteria not met

Remarks:

Not class

Conclusions:

In an acute inhalation study, performed according to OECD guideline 403 and GLP principles, the LC50 of URALAC P 1920C in rats was found to exceed 5 mg/L. Based on these results, the test item is not classified according to GHS and Regulation (EC) No. 1272/2008.

Executive summary:

An acute inhalation study was performed according to OECD/EC guidelines and GLP principles. Five males and five females were exposed to an atmosphere with a time-weighted mean actual concentration of 4.7 ± 0.13 mg/L and observed for 14 days. No mortality occurred. One male showed slow breathing during exposure. One male showed piloerection one hour after exposure. No further clinical signs were noted, no unexpected effects on weight gain were observed and no abnormalities were seen at necropsy. These results show that the the LC50 of URALAC P 1920C in rats exceeds 5 mg/L.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

5 000 mg/m³ air

Quality of whole database:

A reliable study is available (Klimisch 1 study).

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:

19 December 2016 - 03 January 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The study was performed as a data requirement for a registration in China.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Version / remarks:

May 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

August 1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF Guidelines

Version / remarks:

November 2000, including the most recent revisions

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

d.d. 3 November 2015

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

Adjustment was made for specific gravity (1.036) of the vehicle.

In order to obtain homogeneity, the test item (formulations) were heated in a water bath with a maximum temperature of 76.1ºC for a maximum of 38 minutes. The test item (formulations) were allowed to cool to a temperature of maximally 40ºC prior to dosing.

The test item preparations were stirred on a magnetic stirrer during application.

Species:

rat

Strain:

other: Crl: WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant:yes
- Age at study initiation: approx. 10 weeks old
- Weight at study initiation: Males: 258 - 295 g, Females: 191 - 216 g.
- Housing: During the acclimatization period the animals were group housed in Makrolon cages (MIV type, height 18 cm) containing sterilized sawdust as bedding material and paper as cage-enrichment. During the study the animals were individually housed in Makrolon cages (MIII type, height 18 cm.).
- Diet: Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days


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

Type of coverage:

occlusive

Vehicle:

propylene glycol

Remarks:

Specific gravity: 1.036

Details on dermal exposure:

TEST SITE
- Area of exposure: 18 cm^2 for females, 25 cm^2 for males
- % coverage: approx. 10% of the total body surface
- Type of wrap if used: a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing: tap water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 5000 mg/kg bodyweight
- Constant volume or concentration used: yes, 20 mL/kg bodyweight

Duration of exposure:

24 hours

Doses:

5000 mg/kg bodyweight

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations:
- Mortality: twice daily
- Clinical observations: at periodic intervals on the day of dosing and once daily thereafter.
- Body weights: on day 1 (pre-administration), day 8 and day 15.
- Necropsy of survivors performed: yes

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Mortality:

No mortality occurred.

Clinical signs:

other: Red nose was noted for three males and four females on day 2 only.

Gross pathology:

No abnormalities were seen at necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of an acute dermal study (limit study), performed according to OECD/EC guidelines and GLP principles, The dermal LD50 value of URALAC P 1920C in Wistar rats was determined to exceed 5000 mg/kg body weight. As a consequence, URALAC P 1920C is not classified according to GHS and CLP criteria.

Executive summary:

An acute dermal study (limit study) was performed according to OECD/EC guidelines and GLP principles. Five male and five female rats were exposed to 5000 mg/kg bw and observed for 14 days. No mortality occurred. A red nose was noted for three males and four females on day 2 only. No unexpected changes in body weight gain occurred, no abnormalities were seen at necropsy. These reuslts demonstrate that the dermal LD50 value of URALAC P 1920C in Wistar rats exceeds 5000 mg/kg body weight. As a consequence, URALAC P 1920C is not classified according to GHS and CLP criteria.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Quality of whole database:

A reliable study is available (Klimisch 1 study).

Additional information

Oral:

The test substance was assessed for its acute toxicity in an acute oral toxicity study with female Wistar rats, performed according to OECD 423 test guideline and GLP principles (limit test). No mortality occurred. Hunched posture was seen only on the day of treatment for three of the six animals. No abnormalities were found at macroscopic post mortem examination of the animals. Based on the results, the oral LD50 value of URACROSS ZW7672P, Product ID 021116/000 in Wistar rats was established to exceed 2000 mg/kg body weight.

Dermal:

An acute dermal toxicity study with male and female Wistar rats was performed according to the OECD 402 test guideline and GLP principles (limit test). No mortality occurred. Scales, scabs and/or chromodacryorrhoea were noted for three males between Days 1 and 12.

Scales, scabs and/or erythema maculate were noted for all females between Days 5 and 15. The dermal LD50 value of URACROSS ZW7672P, Product ID 021116/000 in Wistar rats was established to exceed 2000 mg/kg body weight.

A second acute dermal study (limit study) was performed according to OECD/EC guidelines and GLP principles. Five male and five female rats were exposed to 5000 mg/kg bw and observed for 14 days. No mortality occurred. A red nose was noted for three males and four females on day 2 only. No unexpected changes in body weight gain occurred, no abnormalities were seen at necropsy. These results demonstrate that the dermal LD50 value of URALAC P 1920C in Wistar rats exceeds 5000 mg/kg body weight.

Inhalation:

An acute inhalation study was performed according to OECD/EC guidelines and GLP principles. Five males and five females were exposed to an atmosphere with a time-weighted mean actual concentration of 4.7 ± 0.13 mg/L and observed for 14 days. No mortality occurred. One male showed slow breathing during exposure. One male showed piloerection one hour after exposure. No further clinical signs were noted, no unexpected effects on weight gain were observed and no abnormalities were seen at necropsy. These results show that the LC50 of URALAC P 1920C in rats exceeds 5 mg/L.

Justification for classification or non-classification

URACROSS ZW7672P, Product ID 021116/000 is not classified for acute toxicity via the oral, dermal and inhalation route and has no obligatory labelling requirement for acute toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.