spirodiclofen (ISO);3-(2,4-dichlorophenyl)-2-oxo-1-oxaspiro[4.5]dec-3-en-4-yl 2,2-dimethylbutyrate

Endpoint summary

Description of key information

An acute oral toxicity study in the rat is available (GLP, draft OECD 423).

An acute inhalation toxicity study in the rat is available (GLP, OECD 403).

An acute dermal toxicity study in the rat is available (GLP, OECD 402).

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:

In life phase: 26 March 1996 - 9 April 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS)
- Source: Harlan Winkelmann GmbH
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8-10 weeks
- Weight at study initiation: 178-183 g (M), 171-174 g (F)
- Fasting period before study: 17 hours
- Housing: group housing by sex
- Diet: ad libitum during the study
- Water: ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5-23.5
- Humidity (%): 40-70
- Air changes (per hr): ~10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 26 March 1996 To: 9 April 1996

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

Carboxymethyl-cellulose Sodium salt

Details on oral exposure:

The test substance was formulated in Carboxymethyl-cellulose Sodium salt 0.5 % before administration. The applied formulations were well mixed and then applied, single administration by gavage to fasted Wistar rats (Hsd Cpb: WU(SPF-bred)): 2000 mg/kg bw, application volume: 10 mL/kg bw.

Doses:

A volume of 10 ml/kg body weight was administered.

No. of animals per sex per dose:

3/sex/group

Control animals:

no

Remarks:

Not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The body weights of the rats are recorded on day 1 before administration and then weekly. Additionally, all animals that died or are sacrificed are weighed.
- Necropsy of survivors performed: yes
- Clinical signs including body weight
- Other examinations performed: clinical signs, body weight, organ weights, histopathology.

Statistics:

Not required

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Key result

Sex:

male/female

Dose descriptor:

LD50 cut-off

Effect level:

2 500 mg/kg bw

Based on:

test mat.

Mortality:

No mortalities.

Clinical signs:

other: other: A dose of 2000 mg/kg bw of Spirodiclofen was tolerated by Wistar rats of both sexes without clinical signs

Gross pathology:

No treatment-related findings.

Findings:

Dose [mg/kg bw]	Toxicological results*			Duration of signs	Time of death	Mortality [%]
male	male			male	male	male
2000	0	0	3	-	-	0
LD50: > 2000 mg/kg bw
Female	Female			Female	Female	Female
2000	0	0	3	-	-	0
LD50: > 2000 mg/kg bw

Interpretation of results:

GHS criteria not met

Conclusions:

Spirodiclofen does not require classification for acute oral toxicity according to the CLP criteria on the basis of this study.

Executive summary:

The acute oral toxicity of spirodiclofen was investigated in the rat, according to the draft OECD 423 guideline.  Groups of male and female Wistar rats (3/sex) were administered single gavage doses of spirodiclofen (in aqueous carboxymethylcellulose) at the limit dose of 2000 mg/kg bw and were observed for 14 days.  There were no deaths, no signs of toxicity and no effects on body weight.  Gross necropsy did not reveal any effects of treatment.  The acute oral LD50 of spirodiclofen was therefore found to be >2000 mg/kg bw under the conditions of this study (OECD 423 LD50 cut-off =2500 mg/kg bw).  Spirodiclofen does not therefore require classification for acute oral toxicity according to the CLP Regulation on the basis of this study.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 2 000 mg/kg bw

Quality of whole database:

An acute oral toxicity study in the rat is available (GLP, draft OECD 423; Klimisch 1).

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:

4 June 1997 - 23 June 1997 (in-life)

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The MMAD at the highest concentration was larger than specified by the test guideline, resulting in a slightly low proportion of particles of respirable size. However, this is stated to be a consequence of attempts to reach the limit concentration.

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EEC 92/69/EEC

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Deviations:

no

Principles of method if other than guideline:

The MMAD at the highest concentration was larger than specified by the test guideline, resulting in a relatively low proportion of particles of respirable size. However, this is stated to be a consequence of attempts to reach the limit concentration.

GLP compliance:

yes (incl. QA statement)

Test type:

traditional method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan-Winkelmann
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 2-3 months
- Fasting period before study: no
- Housing: single
- Diet: ad libitum except during exposure
- Water: ad libitum except during exposure
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): ~50
- Air changes (per hr): ~10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 4 June 1997 To: 23 June 1997

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Remark on MMAD/GSD:

The MMAD at the highest concentration was larger than specified by the test guideline, resulting in a relatively low proprtion of particles of respirable size. However, this is stated to be a consenquence of attempts to reach the limit concentration.

Details on inhalation exposure:

Test material was undiluted, single administration, 4 hours, inhalation dynamic spraying, directed flow nose only. 520-5030 mg/m³ air, aerosol generation conditions: 28 L air /minute.

Analytical verification of test atmosphere concentrations:

yes

Remarks:

The test-substance concentration was determined by gravimetric analysis. The particle-size distribution was analyzed using a critical orifice cascade impactor.

Duration of exposure:

4 h

Remarks on duration:

Standard exposure period according to the test guideline

Concentrations:

520, 5030 mg/m³ air (measured)

No. of animals per sex per dose:

5/sex

Control animals:

yes

Remarks:

Controls were exposed to conditioned air using similar exposure conditions

Details on study design:

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing: Body weights were measured before exposure, on days 3 and 7, and weekly thereafter. Individual weights are also recorded at death, if applicable. The appearance and behavior of each rat were examined carefully several times on the day of exposure and at least once daily thereafter. Weekend assessments were made once a day (morning).

- Necropsy of survivors performed: yes

- Other examinations performed: Rectal temperatures were measured directly after cessation of exposure

Statistics:

Data of rectal temperature measurements were statistically evaluated using ANOVA

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.03 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

There was no mortality

Clinical signs:

other: There were no signs of toxicity

Body weight:

There were no bodyweight effects

Gross pathology:

No treatment-related findings.

Other findings:

Statistical comparison between groups did indicate an effect on body temperature, however, the extent of change is considered to be of no clinical or pathodiagnostic relevance.

Group/Sex	Gravimetric Concentration (mg/L)	Toxicological Result#	Onset and Duration of Signs	Rectal Temperature	Onset of Mortality (%)
1 / m	0	0 / 0 / 5	--	38.0	--
2 / m	0.520	0 / 0 / 5	--	37.9	--
3 / m	5.030	0 / 0 / 5	--	37.8	--
1 / f	0	0 / 0 / 5	--	38.5	--
2 / f	0.520	0 / 0 / 5	--	38.1**	--
3 / f	5.030	0 / 0 / 5	--	38.0*	--

m = males, f = females, -- not applicable; * p ≤ 0.05, ** p ≤ 0.01
# 1st figure = number of dead animals
2nd figure = number of animals with signs
3rd figure = number of animals in the group

Interpretation of results:

GHS criteria not met

Remarks:

Spirodiclofen does not require classification for acute inhalation toxicity according to the CLP criteria, on the basis of this study.

Conclusions:

Spirodiclofen showed no acute toxicity to rats following inhalation administration under the conditions of this study. The acute (4-hour, nose-only) LC50 was found to be >5.03 mg/L,

Executive summary:

The acute inhalation toxicity of spirodiclofen was investigated in a study performed to OECD 403.  Groups of Wistar rats (5/sex) were exposed for four hours (nose only) to analytical concentrations of 0.520 and 5.030 mg/L and observed for 14 days.  There were no mortalities or clinical signs; body weights were unaffected by exposure.  The MMAD at the highest concentration was larger than specified by the test guideline, resulting in a relatively low proportion of particles of respirable size. However, this is stated to be a consenquence of attempts to reach the limit concentration.  The acute (4-hour, nose-only) LC50 of spirodiclofen was found to be >5.03 mg/L,  under the conditions of this study.  Spirodiclofen does not require classification for acute inhalation toxicity acceding to the CLP criteria, on the basis of this study.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

> 5.03 mg/L air

Physical form:

inhalation: dust / mist

Quality of whole database:

An acute inhalation toxicity study in the rat is available (GLP, OECD 403; Klimisch 2).

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:

In-life: 26-March-1996 to 09-April-1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

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: Harlan Winkelmann
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 10 weeks (M), >16 weeks (F)
- Weight at study initiation: 251-269 g (M), 224-243 g (F)
- Fasting period before study: None
- Housing: Single
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5-23.5
- Humidity (%): 40-70
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 26 March 1996 To: 9 April 1996

Type of coverage:

occlusive

Vehicle:

water

Remarks:

sterile demineralized water

Details on dermal exposure:

For each dose and animal, the solid test substance was weighed onto an aluminum foil used to cover the administration site. The test substance was then mixed to a paste with 0.8 ml of sterile demineralized water per gramm test substance and applied to the intact dorsal skin, shorn on the previous day, of 5 rats per dose and sex, respectively. The foil was fixed in place on the skin with an occlusive dressing (fermoflex®). On removal of the dressings the treated skin sites were cleaned with soap and water.

Duration of exposure:

Exposure lasted for 24 hours.

Doses:

2000 mg/kg bw for all test animals

No. of animals per sex per dose:

5 rats per sex per dose

Control animals:

no

Details on study design:

Clinical Observation: Appearance and behavior was recorded several times on the day of treatment, and at least once a day thereafter. Where signs occurred, the type, period and intensity (1 = weak, 2 = moderate, 3 = strong) were determined individually. In the annex they are shown as individual, group and summary findings. Where mortalities occurred, the time recorded was the time the death animal was first noted. On the day of administration, the times reported relate to the time of administration (day 1 of the test). In order to obtain a clearer picture reported times over one hour have been rounded to the nearest full hour. Reported times less than one hour have been rounded to the nearest five minutes. The times reported are the time when the sign first appeared and the time when the sign was last observed. The days recorded are test days.

During clinical observation all abnormal findings were registered and particular attention is paid to the following organ systems, localizations and physiological functions:

Appearance: fur, skin color, edemas, eyes, lacrimation, nasal discharge, salivation etc.
Behavior: grooming, vocalization, excitement, aggression, digging and preening movements, cannibalism etc.
Nervous system: reactivity, motility, reflexes, gait, paralysis, spasms, tremors etc.
Respiration: where assessable, frequency etc.
Cardiovascular system: where assessable, heart rate, pallor etc.
Posture: ventral, lateral recumbency etc.
Gastrointestinal functions: appearance of feces etc.

The post-treatment observation period lasted 14 days. The body weights of the rats are recorded on day 1 before administration and then weekly. Additionally, all animals that died or are sacrificed are weighed. At the end of the post-treatment observation period the animals are anesthetized by inhaling diethyl ether and sacrificed. They are then subjected to a gross pathology examination, as are any animals which may have died intercurrently.

Statistics:

Not required

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

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

Mortality:

There was no mortality

Clinical signs:

other: other: There were no signs of systemic toxicity and no evidence of local dermal irritation at the application site.

Gross pathology:

Gross necropsy did not reveal any effects of treatment.

Interpretation of results:

GHS criteria not met

Conclusions:

Spirodiclofen does not require classification for acute dermal toxicity according to the CLP criteria on the basis of this study.

Executive summary:

The acute oral toxicity of spirodiclofen was investigated in the rat, according to the OECD 403 guideline.  Groups of male and female Wistar rats (5/sex) were administered single dermal doses of spirodiclofen at the limit dose of 2000 mg/kg bw and were observed for 14 days.  There were no deaths, no signs of toxicity or local dermal irritation and no effects on body weight.  Gross necropsy did not reveal any effects of treatment.  The acute dermal LD50 of spirodiclofen was therefore found to be >2000 mg/kg bw under the conditions of this study.  Spirodiclofen does not therefore require classification for acute dermal toxicity according to the CLP Regulation on the basis of this study.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 2 000 mg/kg bw

Quality of whole database:

An acute dermal toxicity study in the rat is available (GLP, OECD 403; Klimisch 1).

Justification for classification or non-classification

Based on the acute oral LD50 (>2000 mg/kg bw), acute inhalation LC50 (>5.03 mg/L) and the acute dermal LD50 (>2000 mg/kg bw) value obtained in GLP- and guideline-compliant studies in the rat, spirodiclofen does not require classification for acute oral, dermal or inhalation toxicity according to CLP criteria.  This is consistent with the harmonised classification and the RAC opinion.  RAC also concluded that In the acute toxicity studies there were no clinical signs of toxicity following oral and dermal exposure.  The decreased core temperature of females following inhalation of spirodiclofen aerosol particles was not accompanied by any other pathological findings and was not regarded relevant for classification. In addition, no effects were observed in the neurotoxicity study. Therefore, there was no clear evidence of specific toxic effects at any target organ or tissue and no signs of respiratory tract irritation or narcotic effects. RAC concluded that no classification for STOT SE was warranted.