Cyclopropanecarboxylic acid, [(3S,4R,4aR,6S,6aS,12R,12aS,12bS)-3-[(cyclopropylcarbonyl)oxy]-1,3,4,4a,5,6,6a,12,12a,12b-decahydro-6,12-dihydroxy-4,6a,12b-trimethyl-11-oxo-9-(3-pyridinyl)-2H,11H-naphtho[2,1-b]pyrano[3,4-e]pyran-4-yl]methyl ester

Endpoint summary

Description of key information

Acute oral toxicity

The LD50 of the acute oral toxicity was determined to be > 2000 mg/kg bw.

 

Acute dermal toxicity

Based on the absence of mortality, the acute dermal LD50 was determined to be > 2000 mg/kg bw.

 

Acute inhalation toxicity

In the acute inhalation toxicity test, no mortality was observed, therefore the LC50 was found to be > 5.48 mg/L.

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:

From January 28, 2009 to October 5, 2009

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF No 12 Nosan No 8147

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

Lot number: 080722
Expiry: 29 July 2010

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Fuji production facility (Shizuoka, Japan)
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 8 weeks at dosing
- Weight at study initiation: 130 – 170 g
- Fasting period before study: 8 weeks at dosing
- Housing: Housed in groups of up to four rats in metal cages with wire mesh floors
- Diet: A certified pellet diet MF (Lot. No. 081015, Oriental Yeast Co., Ltd., Tokyo, Japan), ad libitum
- Water: Local tap water, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2°C
- Humidity: 50 ± 20%
- Air changes (per hr): 10 times or more
- Photoperiod: 12 hours of artificial light (07.00 – 19.00 GMT) in each 24-hour period

IN-LIFE DATES: From 10 Feb 2009 to 13 Mar 2009

Route of administration:

oral: gavage

Vehicle:

methylcellulose

Remarks:

administered in water with 0.5 w/v % methylcellulose solution

Details on oral exposure:

VEHICLE
- Concentration in vehicle: Administered in water with 0.5 w/v % methylcellulose solution
- Lot/batch no.: 080722
- Purity: 95.74%

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

DOSAGE PREPARATION
The test substance formulations were prepared on the day of dosing. The appropriate dose volume of the test substance was administered to each rat by stomach tube.

Doses:

300 mg/kg bw
2000 mg/kg bw

No. of animals per sex per dose:

2 x 3 (two dose groups per dose) females only

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed post dosing at 30 minutes, 3 and 6 hours, and then once daily thereafter until termination. Body weights were taken on days 7 and 14.
- Necropsy of survivors performed: Yes
- Other examinations performed: Macroscopic examination

Statistics:

Not applicable

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred after administration of 300 or 2000 mg/kg bw.

Clinical signs:

other: None

Gross pathology:

Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.

Other findings:

None

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

GLP and guideline compliant

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:

From October 6, 2009 to March 24, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

2009

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: MAFF in Japan, No 12 Nosan No 8147

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

Lot number: 080722
Description: Pale yellow green powder
Expiry: 29 July 2010

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Fuji (Shizuoka, Japan)
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 8 weeks
- Weight at study initiation: 229-247 g (males) and 149-167 g (females)
- Housing: Housed by sex in groups of 5 in holding cages made of stainless steel sheet and wire mesh that was suspended on a moveable rack. (240 mm wide x 400 mm depth x 180 mm height)
- Diet: SDS rat and mouse diet (RM1(E) SQC expanded pellet)
- Water: Tap water supplied by Anglian Water, ad libitum
- Acclimation period: At least 8 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3°C
- Humidity: 50 ± 20%
- Air changes (per hr): at least 100 changes
- Photoperiod (hrs dark / hrs light): 12 hours of artificial light (07.00 – 19.00 GMT) in each 24-hour period

IN-LIFE DATES: From: October 28, 2009 to: November 18, 2009

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose/head only

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

4.3 µm

Geometric standard deviation (GSD):

1.89

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Turn-table type dust feeder with compressed air
- Exposure chamber volume: 31.2 L
- Method of holding animals in test chamber: Snout-only exposure chamber
- Source and rate of air: Airflow, 20 L/min
- Method of particle size determination: Gravimetrically with an electronic balance
- Treatment of exhaust air: Chamber air was exhausted through an air filter system consisting of a bag filter and was emitted to the atmosphere using a blower.
- Temperature, humidity, pressure in air chamber: 22-23°C; 45-51%,- 20 and -5 mm H2O

TEST ATMOSPHERE
- Brief description of analytical method used: HPLC
- Samples taken from breathing zone: Yes, 6 L of chamber air was drawn from the chamber sampling port using an air sampler

TEST ATMOSPHERE
- Particle size distribution: Measured three times with an Andersen type personal sampler at 1, 2 and 3 hours after the initiation of exposure.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

5.48 mg/L

No. of animals per sex per dose:

5

Control animals:

yes

Remarks:

a vehicle control group (1.31 mg/L for white carbon) was conducted with 5 males and 5 females.

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of weighing: Shortly before exposure and on 1, 3, 7 and 14 days after exposure
- Necropsy of survivors performed: Yes
- Other examinations: All animals were checked at 2 hours during the exposure. Further clinical observations were made immediately post exposure as well as 1 and 4 hours post exposure and daily thereafter.

Statistics:

Statistical methods employed are not detailed in the final report.

Preliminary study:

Not applicable

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.48 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

None

Clinical signs:

other: In the test substance group, abnormal respiratory sound was noted in four males and one female. This sign began to appear immediately after the termination of exposure and disappeared by 4 hours after the termination of exposure. No signs were noted in an

Body weight:

Several males in test substance group and all animals in vehicle control group lost body weight on day 1 after exposure. On 3 days afer exposure, all animals in vehicle control group lost body weight. All animals gained body weight by 7 and 14 days after exposure.

Gross pathology:

No gross pathological abnormalities were detected in the animals that underwent necropsy at termination of the study.

Other findings:

No other findings were observed.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating conc.

Value:

5 480 mg/m³ air

Quality of whole database:

GLP and guideline compliant

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:

From January 28, 2009 to October 5, 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF No 12 Nosan No 8147

Version / remarks:

1987

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

Lot number: 080722
Expiry: 29 July 2010

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Fuji (Shizuoka, Japan).
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 9 weeks at dosing
- Weight at study initiation: 271- 283 g (male); 170-180 g (female)
- Fasting period before study: A fasting period was not reported
- Housing: Housed individually in metal cages (310 mm wide x 410 mm depth x 230 mm height)
- Diet: Scertified pellet diet MF (Lot No 081015, Oriental Yeast Co; Tokyo, Japan), ad libitum
- Water: Tap water, ad libitum
- Acclimation period: 9 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2°C
- Humidity: 50 ± 20%
- Air changes: 10x or more per hour
- Photoperiod: 12 hours of artificial light (0700 – 1900 GMT) in each 24-hour period

IN-LIFE DATES: From Feb 19, 2009 to March 5, 2009

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: Dorso-lumbar region; about 26hr prior to treatment, hair was removed with electric clippers, on the dorsal body surface area.
- Type of wrap: Surgical dressing pad moistened with 0.5 mL of distilled water, then applied to the 4 cm X 5 cm clipped skin area and occlusively held in place with surgical tape (Transpore TM).

REMOVAL OF TEST SUBSTANCE
- Washing: Residual test substance was washed off with warm water and neutral detergent
- Time after start of exposure: After 24 hours

TEST MATERIAL
- Concentration: 66.7% in 1% w/v aqueous methylcellulose
- Constant volume or concentration used: Yes

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

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: Animals were carefully observed for their mortalities, appearance, posture, behavior, respiration, consciousness, neurologic signs, body temperature, excretion, etc., at the time points of 1, 3 and 6 hours after administration and once daily thereafter until the termination of the observation period. Each animal was weighed on day of the administration shortly before dosing and on days 7 and 14 after administration.
- Necropsy of survivors performed: Yes
- Other examinations performed: Macroscopic examination consisted of opening the thoracic and abdominal cavities. The macroscopic appearance of all tissues including the dose sites was recorded.

Statistics:

None

Preliminary study:

Not applicable

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed in this study.

Clinical signs:

other: No systemic clinical signs were observed during the study. There were no observed dermal responses.

Gross pathology:

Macroscopic examination revealed no abnormal findings in any of the animals at the termination of the study.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

GLP and guideline compliant

Additional information

Key study: Acute oral toxicity 2010/1231357

In an acute oral toxicity study, three young adult female Wistar (Jcl:Wistar) rats were given an oral dose via gavage of the test substance suspended in 0.5 w/v% methylcellulose solution at a dose levels of 300 mg/kg (two dose groups of 3 animals each) and 2000 mg/kg bw (two dose groups of 3 animals each) (Batch: 080722; Purity: 95.74%). Animals were observed for 14 days.

After administration of 300 mg/kg (first and second step), 2000 mg/kg bw (third and fourth step) of the test substance, all animals survived until the termination of the study. Accordingly, the oral LD50 was found to be greater than 2000 mg/kg bw. No clinical signs were noted in any of the dose groups in the study. All animals gained body weight on days 7 and 14 after administration. No macroscopic abnormalities were noted in any animal at necropsy at the end of the observation period.

Accordingly, the oral LD50 was > 2000 mg/kg bw.

 

Key study: Acute dermal toxicity 2009/1130542

In an acute dermal toxicity study, one group of 5 male and 5 female Wistar rats were exposed to a single dermal dose of 2000 mg/kg of the test substance (Batch:080722; Purity: 95.74%) to the clipped, dorsal skin area under semi-occlusive conditions for 24 hours. The animals were observed for 14 days after administration. There were no dermal responses, clinical signs, macroscopic abnormalities or altered bodyweight parameters.

Based on the absence of mortality, the acute dermal LD50 was determined to be > 2000 mg/kg bw.

 

Key study: Acute inhalation toxicity 2010/1231352

Wistar rats (5 males and 5 females) were exposed to the test substance for 4 hours at a concentration of 5.48 mg/l (maximum attainable concentration; nose only) and were observed for 14 days. The mass median aerodynamic diameter (MMAD) of the test aerosol was 4.30 μm. A vehicle control group (1.31 mg/L for white carbon) was conducted with 5 males and 5 females.

With no observed mortality, the LC50 was found to exceed 5.48 mg/l.

Justification for classification or non-classification

Acute oral toxicity

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. The LD50 was greater than 2000 mg/kg bw. As a result the substance is not considered to be classified for acute oral toxicity under Regulation (EC) No 1272/2008, as amended for the eighth time in Regulation (EU) No 2016/918.

Acute dermal toxicity

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. The LD50 was greater than 2000 mg/kg bw. As a result the substance is not considered to be classified for acute dermal toxicity under Regulation (EC) No 1272/2008, as amended for the eighth time in Regulation (EU) No 2016/918.

Acute inhalation toxicity

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. The LC50 was greater than 5.48 mg/L. As a result the substance is not considered to be classified for acute inhalation toxicity under Regulation (EC) No 1272/2008, as amended for the eighth time in Regulation (EU) No 2016/918.