3-(methylthio)propionaldehyde

Endpoint summary

Description of key information

Oral (similar to OECD 401), rat (m): LD50 = 490 mg/kg bw

Inhalation (similar to OECD 403), rat (m, f): LC50 = 4.8 mg/L air

Dermal (similar to OECD 402), rabbit (m, f): LD50 = 736 mg/kg bw (males) and 818 mg/kg bw (females)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

No information on purity of the test material, brief description, only 5 rats/dose level, 1-4 animals of the same sex/dose level, no details given on animals, duration of observation period not specified, no data on individual animals

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

- Principle of test: Acute oral toxicity in rats.
- Short description of test conditions: The test item was applied by stomach tube as a 50% dilution in corn oil. Groups of 1-4 animals per dose were used to determine the minimum lethal dose and to calculate the LD50.
- Parameters analysed / observed: Clinical signs of toxicity, mortality and gross necropsy.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

corn oil

Remarks:

50% solution

Doses:

1750, 2000, 2200 and 2500 mg/kg bw

No. of animals per sex per dose:

1750 mg/kg bw: 2 males and 3 females
2000 mg/kg bw: 3 males and 3 females
2200 mg/kg bw: 1 male and 4 females
2500 mg/kg bw: 3 males and 2 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: Not specified, survival time was 8 to 36 hours.
- Necropsy of survivors performed: yes

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

2 300 mg/kg bw

Based on:

test mat.

Mortality:

1750 mg/kg bw: 1/2 males and 0/3 females died.
2000 mg/kg bw: 0/3 males and 2/3 females died.
2200 mg/kg bw: 0/1 male and 2/4 females died.
2500 mg/kg bw: 2/3 males and 2/2 females died.
Survival time was 8 to 36 hours with most deaths occuring over night.

Clinical signs:

Symptoms of toxicity included discomfort followed by lethargy, salivation and weakness.

Gross pathology:

At autopsy there was pulmonary hyperemia and inflammation of the gastric mucosa.

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

CLP: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1974-02 to 1974-04

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

No information on purity of the test substance, no data of individual animals reported, no information on body weights, mortality rate based on 7 days, observation period 4 weeks

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

No information on purity of the test substance, no data of individual animals reported, no information on body weights, mortality rate based on 7 days, observation period 4 weeks

Principles of method if other than guideline:

- Principle of test: Determination of oral LD50 in rats
- Short description of test conditions: The test item was applied as received by oral gavage to groups of 10 male and 10 female rats per dose. The animals were observed for a period of 4 weeks prior to necropsy, mortality rates were determined on a 7-day observation period.
- Parameters analysed / observed: Behavior, food and water consumption, body weight development and mortality.

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Details on test animals or test system and environmental conditions:

- Source: S.IVANOVAS, Kissleg, Germany
- Age at study initiation: 38 days (males) and 42 days (females)
- Weight at study initiation: 100 – 105 g
- Fasting period before study: yes
- Housing: individually in Makrolon type II cages
- Diet: ALTROMIN 1323 (Altromin GmbH, Lage, Germany), ad libitum
- Water: tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 0.5
- Humidity (%): 60 ± 3

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION: The test substance was applied as received (no vehicle). Amounts applied were based on the animal's bodyweight.

Doses:

3.18, 4.00, 5.04, 6.35, 7.90 and 10 mL/kg bw corresponding to 3294, 4144, 5221, 6579, 8184 and 10360 mg/kg bw.

No. of animals per sex per dose:

10

Control animals:

no

Details on study design:

- Duration of observation period following administration: 4 weeks
- Frequency of observations and weighing: not specified
- Necropsy of survivors performed: yes
- Other examinations performed: food and water consumption were monitored

Statistics:

The LD50 was calculated according to Lichtfeld and Wilcoxon, based on a mortality rate within 7 days.

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

5 926 mg/kg bw

Based on:

test mat.

95% CL:

>= 5 335 - < 6 568

Remarks on result:

other: corresponding to 5.72 mL/kg bw (95% CL: 5.15 - 6.34 mL/kg bw), dosel levels in mg/kg bw calculated with a density of 1036 mg/mL

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

6 579 mg/kg bw

Based on:

test mat.

95% CL:

>= 5 926 - < 6 568

Remarks on result:

other: corresponding to 6.35 mL/kg bw (95% CL: 5.72 - 6.34 mL/kg bw), dosel levels in mg/kg bw calculated with a density of 1036 mg/mL

Mortality:

3294 mg/kg bw (corresponding to 3.18 mL/kg bw): No mortality occurred.
4144 mg/kg bw (corresponding to 4.00 mL/kg bw): No mortality occurred.
5221 mg/kg bw (corresponding to 5.04 mL/kg bw): 2/10 males and 3/10 females died (Days 2 and 3 post-dosing).
6579 mg/kg bw (corresponding to 6.35 mL/kg bw): 5/10 males and 4/10 females died (Days 2 and 3 post-dosing).
8184 mg/kg bw (corresponding to 7.9 mL/kg bw): 6/10 males and 6/10 females died (Days 2 and 3 post-dosing).
10360 mg/kg bw (corresponding to 10.0 mL/kg bw): 10/10 males and 10/10 females died (12 h - 4 Days post-dosing).

Clinical signs:

3294 mg/kg bw (corresponding to 3.18 mL/kg bw): No clinical signs of toxicity were observed.
4144 mg/kg bw (corresponding to 4.00 mL/kg bw): post dosing slight sedation and ataxia, slight mydriasis, decreased food consumption on Day 1, all animals appeared normal on study Day 2.
5221 mg/kg bw (corresponding to 5.04 mL/kg bw): post dosing moderate sedation and ataxia, hypnoe, slight mydriasis, decreased food consumption on Days 1 and 2, effects were fully reversible in surviving animals up to study Day 3, ante-mortem signs in moribund animals: coma.
6579 mg/kg bw (corresponding to 6.35 mL/kg bw): post dosing slight to moderate sedation and ataxia, hypnoe, slight mydriasis, decreased food consumption on Days 1 and 2, effects were fully reversible in surviving animals up to study Day 3, ante-mortem signs in moribund animals: coma.
8184 mg/kg bw (corresponding to 7.9 mL/kg bw): post dosing slight to moderate sedation and ataxia, hypnoe, slight mydriasis, decreased food consumption on Days 1 and 2, effects were fully reversible in surviving animals up to study Day 3, ante-mortem signs in moribund animals: coma.
10360 mg/kg bw (corresponding to 10.0 mL/kg bw): post dosing increasing sedation and ataxia, hypnoe and mydriasis, coma in all rats within 6 - 24 h post-dosing.

Body weight:

No data

Gross pathology:

3294 mg/kg bw (corresponding to 3.18 mL/kg bw): gross necropsy revealed no abnormal findings.
4144 mg/kg bw (corresponding to 4.00 mL/kg bw):: gross necropsy revealed no abnormal findings.
5221 mg/kg bw (corresponding to 5.04 mL/kg bw): hematoma in the thymus in dead animals, surviving animals without abnormal findings.
6579 mg/kg bw (corresponding to 6.35 mL/kg bw): hematoma in the thymus in dead animals, surviving animals without abnormal findings.
8184 mg/kg bw (corresponding to 7.9 mL/kg bw): hematoma in the thymus in dead animals, surviving animals without abnormal findings.
10360 mg/kg bw (corresponding to 10.0 mL/kg bw): hematoma in the thymus in dead animals.

Interpretation of results:

GHS criteria not met

Conclusions:

CLP: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

only short summary available

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

adopted 12 May 1981

Deviations:

yes

Remarks:

5 rats per dose level (2-3 male, 2-3 female), no details on animals and environmental conditions, body weight not recorded

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Doses:

501, 631, 794 and 1000 mg/kg bw

No. of animals per sex per dose:

501 mg/kg bw: 3 males and 2 females
631 mg/kg bw: 2 males and 3 females
794 mg/kg bw: 3 males and 2 females
1000 mg/kg bw: 2 males and 3 females

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days

Sex:

male/female

Dose descriptor:

LD50

Effect level:

700 mg/kg bw

95% CL:

600 - 820

Mortality:

501 mg/kg bw: 0/3 males and 1/2 females died within several hours up to one day post-administration.
631 mg/kg bw: 1/2 males and 1/3 females died within several hours up to one day post-administration.
794 mg/kg bw: 2/3 males and 2/2 females died within several hours up to one day post-administration.
1000 mg/kg bw: 2/2 males and 2/3 females died within several hours up to one day post-administration.

Clinical signs:

Reduced appetite and activity (one day in survivors) rapidly increasing weakness, collapse and death.

Gross pathology:

Lung and liver hyperemia and gastrointestinal inflammation were noted in decedent animals.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute oral LD50 was determined to be 700 mg/kg bw for male and female rats.
CLP: Acute Oral 4 (H302) according to Regulation (EC) No. 1272/2008

Reference 4

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

adopted 12 May 1981

Deviations:

yes

Remarks:

No information on purity of the test substance.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Centre d'élevange IFFA CREDO, France
- Weight at study initiation: 160 - 180 g
- Housing: 5 rats/cage in cages measuring 37.5 x 23.5 x 16 cm with sawdust bedding, sterilized and free of dust
- Diet: IFFARAT, except for the last 18-20 hours prior treatment
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1
- Humidity (%): 50 ± 10
- Air changes (per hr): 8

Route of administration:

oral: gavage

Vehicle:

other: 10% aqueous dispersion of arabic gum

Details on oral exposure:

VEHICLE
- Concentration in vehicle: Due to the results of the preliminary test, the doses for the main test were prepared by 5% dilution of test item in the vehicle as described above (10% arabic gum). The 100 and 500 mg/kg bw dose levels were prepared by 5% dilution in the vehicle

MAXIMUM DOSE VOLUME APPLIED: 20 mL/kg bw

Doses:

400, 500, 700, 800 and 1000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: immediately after application, then after 1, 2 and 6 h post dosing, and daily thereafter
- Frequency of weighing: on study Days 0, 1, 2, 4, 7 and 14
- Necropsy of survivors performed: yes

Statistics:

The LD50 was calculated according to Lichtfeld and Wilcoxon.

Preliminary study:

In a preliminary test, the following dose levels were investigated: 100, 500, 1000, 2500 and 5000 mg/kg bw. For the 2 lowest dose levels, the test item was administered as 5% aqueous solution, at an application volume of respectively 2 and 10 ml/kg bw. For the higher dose levels, the test item was administered as such, i.e., undiluted. No mortality was noticed at the lowest tested dose of 100 mg/kg bw. At 500 mg/kg bw, one of 2 males died. At 1000 mg/kg bw and above, mortality was 100% in both sexes. Based on the results of the study, the vehicle and test doses for the main study were selected.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

755 mg/kg bw

Based on:

test mat.

95% CL:

>= 654 - < 872

Mortality:

400 mg/kg bw: no mortality occurred.
500 mg/kg bw:1/5 males died within 2 h post-administration; all females survived.
700 mg/kg bw: 3/5 males died (1 and 3 h post-administration); all females survived.
800 mg/kg bw: 4/5 males and 3/5 females died (1 and 2 h post-administration).
1000 mg/kg bw: 5/5 males and 3/5 females died (1 and 2 h post-administration).
In the control group no mortality occurred.

Clinical signs:

At 400 mg/kg bw and above, reduced activity, apathy and slight piloerection were observed from 1 h up to 6 h after treatment. Additionally, at 800 and 1000 mg/kg bw tremors were noted from 1 h up to 6 h after treatment. All surviving animals fully recovered within 24 h post-treatment.

Body weight:

400 mg/kg bw: body weight was reduced in males and females over the whole study period, reaching statistically significance for males on study Days 7 and 14.
500 mg/kg bw: body weight was reduced in males (not significant) over the whole study period.
700 mg/kg bw: body weight was reduced in females over the whole study period, reaching statistical significance from study Day 1 onwards.

Gross pathology:

Haemorrhage of the lungs was noted in all decedent animals and haemorrhage of the gastrointestinal tract was noted in single animals. In animals that were sacrificed at the end of the observation period the only finding reported was a dark discoloration of the spleen at 1000 mg/kg bw.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

CLP: Acute Oral 4, H302 according to Regulation (EC) No. 1272/2008 (CLP/EU GHS)

Reference 5

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

no detailed information, only 2-4 rats/dose level, duration of observation period not specified, no body weight data reported, no data of individual animals, no necropsy performed

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Research study, no guideline followed.
- Principle of test: Determination of oral LD50 in rats.
- Short description of test conditions: The test item was applied as received by oral gavage to groups of 2-4 male rats per dose.
- Parameters analysed / observed: mortality, clinical signs and body weight development.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River
- Mean weight at study initiation: 289 g
- Diet: ad libitum
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°F): 75 ± 2
- Humidity (%): 55
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DOSAGE PREPARATION: The test item was administered as received (unchanged).

Doses:

0.65, 1.29, 2.6, 3.5 and 5.23 mL/kg bw, corresponding to 673, 1336, 2694, 3626 and 5418 mg/kg bw.

No. of animals per sex per dose:

673 mg/kg bw (corresponding to 0.65 mL/kg bw): 2 males
1336 mg/kg bw (corresponding to 1.29 mL/kg bw): 2 males
2694 mg/kg bw (corresponding to 2.6 mL/kg bw): 3 males
3626 mg/kg bw (corresponding to 3.5 mL/kg bw): 3 males
5418 mg/kg bw (corresponding to 5.23 mL/kg bw): 4 males

Control animals:

yes

Sex:

male

Dose descriptor:

LD50

Effect level:

1 336 mg/kg bw

Based on:

test mat.

Remarks on result:

other: corresponding to 1.29 mL/kg bw, dosel levels in mg/kg bw calculated with a density of 1036 mg/mL

Mortality:

673 mg/kg bw (corresponding to 0.65 mL/kg bw): 1/2 males died (within 3 h post-dosing).
1336 mg/kg bw (corresponding to 1.29 mL/kg bw): 1/2 males died (within 3 h post-dosing).
2694 mg/kg bw (corresponding to 2.6 mL/kg bw): 3/3 males died (within 2 h post-dosing).
3626 mg/kg bw (corresponding to 3.5 mL/kg bw): 3/3 males died (within 2 h post-dosing).
5418 mg/kg bw (corresponding to 5.23 mL/kg bw): 4/4 males died (5-17 minutes post-dosing).

Clinical signs:

673 and 1336 mg/kg bw (corresponding to 0.65 and 1.29 mL/kg bw): convulsion immediately after dosing, sedatation and body weight loss on study day 1, all surviving rats appeared normal from study day 2 onwards.
2694, 3626 and 5418 mg/kg bw (corresponding to 2.6, 3.5 and 5.23 mL/kg bw): ante-mortem signs were sedation and ataxia within 35 minutes post-dosing, audible and labored breathing.

Body weight:

673 and 1336 mg/kg bw (corresponding to 0.65 and 1.29 mL/kg bw): body weight loss in surviving rats on study Day 1, from Day 2 onwards all surviving rats fully recovered

Interpretation of results:

study cannot be used for classification

Conclusions:

The available data are not sufficient to evaluate the test items potential for acute oral toxicity as the number of animals used was too low.

Reference 6

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1985-08-20 to 1985-11-07

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

adopted 12 May 1981

Deviations:

yes

Remarks:

no information on analytical purity

Qualifier:

according to guideline

Guideline:

other: FIFRA Pesticide Assessment Guidelines, Subdivision F, Hazard Evaluation: Human and Domestic Animals; Office of Pesticide Programs, U.S. Environmental Protection Agency, Office of Pesticides and Toxic Substances, November 1982; Section 81-1, Acute Oral Tox

Qualifier:

according to guideline

Guideline:

other: TSCA: Health Effects Test Guidelines, Office of Toxic Substances, Office of Pesticides and Toxic Substances, United States Environmental Protection Agency, August 1982 Acute Exposure, Oral Toxicity

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: CDR (Sprague-Dawley derived) Albino Rats

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test animals
- Source: Charles River Breeding Laboratories, Inc., Wilmington, USA
- Age at study initiation: approx. 9 - 12 wks old; females were nulliparous and non-pregnant
- Weight at study initiation: 231 - 318 g (males) and 215 - 227 g (females)
- Acclimation period: 8, 15 or 16 days
- Fasting over-night (approx. 18 h) prior to treatment
- Housing: Grouped housing (six/cage) during equilibration. Individually housed during study.
- Diet: Purina Laboratory Chow, 5001; ad libitum
- Water: Municipal water; ad libitum

Environmental conditions
- Temperature (°C): 19 - 24
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DOSAGE PREPARATION: The test material was administered as received, no preparation was necessary. Animals received the same concentration of the undiluted test substance. Body burdens were adjusted via the applied volume.

Doses:

Males: 250, 350, 500, 600 and 700 mg/kg bw
Females: 700, 850, 1000, 1200 and 1700 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: approximately 1, 2 and 4 h after dosing and daily thereafter
- Frequency of weighing: pre-fasting, post-fasting (weights used for calculation of doses), Day 7 and Day 14
- Necropsy of survivors performed: Yes, all animals surviving at termination of the post exposure observation period were sacrificed and examined grossly. All abnormalities were recorded but no tissues were saved.
- Other examinations performed: Yes, food consumption was observed in the 14-days observation period.

Statistics:

- The LD50 was estimated with 95% confidence limits by calculation according to Miller, Lloyd C. & Tainter, M.L.
- Source: Estimation of the ED50 and its errors by means of logarithmic-probit graph paper; Proc. Soc. Exp. Bio. Med 57: 261-264 (1944)

Preliminary study:

A range-finding experiment was conducted with one male and one female rat per dose.
- The doses tested were 350, 500, 700, 1000, and 1500 mg/kg bw.
- The post exposure observation time was 7 days.
- One male animal died after exposure to >= 700 mg/kg bw.
- The female rats died at doses of 1000 and 1500 mg/kg bw.

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

490 mg/kg bw

Based on:

test mat.

95% CL:

>= 402 - <= 578

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

1 050 mg/kg bw

Based on:

test mat.

95% CL:

>= 854 - <= 1 246

Mortality:

250 mg/kg bw (males only): No mortality occurred.
350 mg/kg bw (males only): No mortality occurred.
500 mg/kg bw (males only): 4/5 males died (1 h - 2 days post-dose).
600 mg/kg bw (males only): 2/5 males died (2 h post-dose).
700 mg/kg bw: 5/5 males and 1/5 females died (2 - 4 h post-dose).
850 mg/kg bw (females only): 1/5 females died (2 h post-dose).
1000 mg/kg bw (females only): 2/5 females died (2 h post-dose).
1200 mg/kg bw (females only): 5/5 females died (1 - 4 h post-dose).
1700 mg/kg bw (females only): 4/5 females died (1 - 24 h post-dose).

Clinical signs:

≥ 250 mg/kg bw: oral, ocular and nasal discharge, hypopnea, dyspnoea, urinary staining or hypoactivity after dosing in males and females and decreased food consumption on study Days 1 and 2, effects reversible in surviving animals by study Day 3.
≥ 500 mg/kg bw: ataxia, dry and wet rales, prostration, eyes partially closed, ocular discharge after dosing in males and females and decreased food consumption up to study Day 4, effects reversible in surviving animals by study Days 4 - 5.

Body weight:

The majority of surviving animals had gained weight both 7 and 14 days after dosing.
1700 mg/kg bw: Body weight loss at Day 7 in the single surviving female.

Gross pathology:

250 and 350 mg/kg bw: No abnormal findings.
500 mg/kg bw: discoloration of lungs in 3/4 dead males, testes in body cavity of 2/4 dead males, red walls of stomach in 1/4 dead males, red walls + red fluid in intestine in 4/4 dead males.
600 mg/kg bw: discoloration of lungs in 2/2 dead males, testes in body cavity of 1/2 dead males, red walls + red fluid in intestine in 2/2 dead males.
700 mg/kg bw: discoloration of lungs in 5/5 dead males, testes in body cavity of 1/5 dead males, red walls of stomach in 1/5 dead males, test material in stomach in 2/5 dead males, red walls + red fluid in intestine in 5/5 dead males; uterus reddened in 1/4 and uterus swollen in 1/4 surv. females.
850 mg/kg bw: discoloration of lungs in 1/1 dead females, reddened ovaries in 2/4 surv. females, uterus reddened in 2/4 surv. females, red walls + fluids of the intestine in 1/1 dead females.
1000 mg/kg bw: discoloration of lungs in 2/2 dead females, reddened uterus in 1/3 surv. females, swollen uterus in 1/3 surv. females, red walls of stomach in 1/2 dead females, red walls + fluids of the intestine in 2/2 dead females.
1200 mg/kg bw: red foci in lungs of 2/5 dead females, discoloration of lungs in 5/5 dead females, swollen uterus in 2/5 dead females, red walls and wrinkled stomach each in 1/5 dead females, test material content in the stomach of 4/5 dead females, white precipitate in the stomach of 3/5 dead females, red walls of the intestine in 1/5 dead females, contents of test material in the intestine in 2/5 dead females.
1700 mg/kg bw: post-mortem internal autolytic changes in 1/4 dead females, red foci of lungs in 1/4 dead females, discoloration of lungs in 4/4 dead females, swollen uterus in 1/4 dead females, red walls of stomach in 2/4 dead females, contents of test material in the stomach of 4/4 dead females, white precipitate in the stomach of 3/4 dead females, red fluid of the intestine in 2/4 dead females, contents of test material in the intestine in 1/4 dead females.

Other findings:

Food consumption:
Most animals had decreased food consumption beginning on the day after dosing. Food consumption recovered back to normal by study Day 5 onwards.

- Changes in surviving animals sacrificed after 14 days were compared in the study report to those of control animals. However, no other reference is made to control animals in this study.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The oral LD50 for male rats was determined to be 490 mg/kg bw and that of female rats to be 1050 mg/kg bw.
CLP: Acute Oral 4 (H302) according to Regulation (EC) No. 1272/2008

Reference 7

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

adopted 12 May 1981

Deviations:

yes

Remarks:

test item administered as 25% formulation in corn oil, no details on animals and environmental conditions

Principles of method if other than guideline:

An acute peroral toxicity study was conducted in Sprague Dawley rats. The test item was administered by oral gavage as a 25% formulation diluted in corn oil. Each 5 males or 3-5 females received a single oral dose of 2.0, 1.0 and 0.5 mL/kg bw (males) or 4.0, 2.0, 1.0, 0.5 and 0.25 mL/kg bw (females). The animals were inspected daily up to 14 days post dosing for signs of toxic and pharmacological effects. Body weights were recorded prior to administration and 7 and 14 days post-dosing. All animals that died or survived until study termination were necropsied and examined for any signs of gross pathology.

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

corn oil

Remarks:

25% formulation (v/v)

Details on oral exposure:

VEHICLE
- Amount of vehicle: 0.25, 0.5, 1.0, 2.0 and 4.0 mL/kg bw

MAXIMUM DOSE VOLUME APPLIED: 4.0 mL/kg bw

Doses:

0.5, 1.0 and 2.0 mL/kg bw in males, corresponding to 518, 1036 and 2072 mg/kg bw; and 0.25, 0.5, 1.0, 2.0 and 4.0 mL/kg bw in females, corresponding to 259, 518, 1036, 2072 and 4144 mg/kg bw.

No. of animals per sex per dose:

259 mg/kg bw (corresponding to 0.25 mL/kg bw): 3 females
518 mg/kg bw (corresponding to 0.5 mL/kg bw): 5 males and 3 females
1036 mg/kg bw (corresponding to 1.0 mL/kg bw): 5 males and 5 females
2072 mg/kg bw (corresponding to 2.0 mL/kg bw): 5 males and 5 females
4144 mg/kg bw (corresponding to 4.0 mL/kg bw): 5 females

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: daily
- Frequency of weighing: before dosing, 7 and 14 days post-dosing
- Necropsy of survivors performed: yes

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

1 036 mg/kg bw

Based on:

test mat. (dissolved fraction)

95% CL:

611 - 1 761

Remarks on result:

other: corresponding to 1.0 mL/kg bw (95% CL: 0.59 - 1.7 mL/kg bw), dosel levels in mg/kg bw calculated with a density of 1036 mg/mL

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

1 740 mg/kg bw

Based on:

test mat. (dissolved fraction)

95% CL:

984 - 3 098

Remarks on result:

other: corresponding to 1.68 mL/kg bw (95% CL: 0.95 - 2.99 mL/kg bw), dosel levels in mg/kg bw calculated with a density of 1036 mg/mL

Mortality:

259 mg/kg bw (corresponding to 0.25 mL/kg bw): no mortality was observed.
518 mg/kg bw (corresponding to 0.5 mL/kg bw): no mortality was observed.
1036 mg/kg bw (corresponding to 1.0 mL/kg bw): 3/5 males and 1/5 females died within 3.5 - 4 h post-administration.
2072 mg/kg bw (corresponding to 2.0 mL/kg bw): 4/5 males and 3/5 females died within 2 h up to 1 day post-administration.
4144 mg/kg bw (corresponding to 4.0 mL/kg bw): 5/5 females died 1.5 - 4 h post-administration.

Clinical signs:

1036, 2072 and 4144 mg/kg bw (corresponding to 1.0, 2.0 and 4.0 mL/kg bw): sluggishness, salivation, lacrimation, unsteady gait and laboured breathing were seen within 5-15 minutes and prostration within 1 hour post-dosing. Survivors recovered from these effects within 2 days after dosing.

Body weight:

Surviving animals gained body weight as expected.

Gross pathology:

Mottled and red lungs, liquid filled stomachs, red liquid-filled intestines and discolorations of kidneys was noted in all decedent animals. There were no macroscopic findings in surviving animals.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute oral LD50 was determined to be 1036 mg/kg bw for male rats and 1740 mg/kg bw for female rats.
CLP: Acute Oral 4 (H302) according to Regulation (EC) No. 1272/2008

Reference 8

Endpoint:

acute toxicity: oral

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

data reported as part of a review, no experimental details given

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

No details on materials and methods were reported.

GLP compliance:

not specified

Species:

mouse

Dose descriptor:

LD50

Effect level:

1 620 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50: 925 ± 2830 mg/kg bw

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute oral LD50 was 1620 (925 ± 2830) mg/kg bw for mice.
CLP: Acute Oral 4 (H302) according to Regulation (EC) No. 1272/2008

Reference 9

Endpoint:

acute toxicity: oral

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

data reported as part of a review, no experimental details given

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

No details on materials and methods were reported.

GLP compliance:

not specified

Species:

rat

Dose descriptor:

LD50

Effect level:

4 400 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 3390 ± 5720 mg/kg bw

Dose descriptor:

LD100

Effect level:

5 920 mg/kg bw

Based on:

test mat.

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

The acute oral LD50 was 4400 (3390 ± 5720) mg/kg bw for rats.
CLP: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

490 mg/kg bw

Quality of whole database:

The available information comprises adequate and reliable studies, and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC) No. 1907/2006.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

(no details given, dose unclear, no analytical determinations, no particle size determination, mass median aerodynamic diameter not given, number of animals too low, observation period of 5 days only, body weight was not recorded, gross necropsy of one animal only)

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Four mature male rats were exposed in a metal drum of approximately 25 L capacity and exposed for 4 h to an atmosphere containing vapors of the test item.
- Short description of test conditions: The vapors entered the chamber through a glass tube attached to a 500 mL boiling flask with an opening at both ends. The flask was placed in a horizontal position and approximately one ounce of Propionaldehyde added. Air from a compressor was introduced through one end by means of a glass tube inserted in a rubber stopper. Air pressure of two pounds per square inch gently rippled the fluid in the curvature of the flask but did not force droplets up into the tube leading to the chamber. As a precaution a piece of 200 mesh bolting cloth doubled was attached to the end of the tubs entering the chamber to prevent any possible carryover of atomised liquid and yet allow vapors to pass.
- Parameters analysed / observed: Periodic examination of the animals for clinical signs and mortality was made by flashlight through a glass cover placed over the drum. Average relative humidity and temperature in the chamber were determined.

GLP compliance:

no

Test type:

traditional method

Limit test:

yes

Species:

rat

Strain:

not specified

Sex:

male

Route of administration:

inhalation: dust

Type of inhalation exposure:

whole body

Vehicle:

air

Remarks:

approx. one ounce of propionaldehyde was added

Analytical verification of test atmosphere concentrations:

no

Duration of exposure:

4 h

No. of animals per sex per dose:

4

Control animals:

no

Details on study design:

- Duration of observation period following administration: 5 days
- Necropsy of survivors performed: yes

Mortality:

No mortality was observed.

Clinical signs:

other: For details please refer to "Any other information on results".

Gross pathology:

One animal was sacrificed for macroscopic examination of the viscera. Numerous haemorrhagic areas were present in the lungs.

Clinical signs of toxicity: Restlessness was observed shortly after the beginning of exposure; excessive lacrimation and salivation were evident 5 -10 minutes after start of exposure, bloody nasal discharge appeared after 10-15 minutes of exposure; reduced activity with increasing exposure time; irregular breathing developing into gasping during the latter half of the exposure period, 2/4 males showed bronchal rales immediately after exposure.

Interpretation of results:

study cannot be used for classification

Reference 2

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

only short summary

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: 6 male Sprague-Dawley rats were exposed to 3.9 mg/L test item via inhalation exposure for 5.5 h.
- Short description of test conditions: No experimental details given.
- Parameters analysed / observed: clinical signs, mortality, gross necropsy of decedents.

GLP compliance:

not specified

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Duration of exposure:

5.5 h

Concentrations:

3.9 mg/L

No. of animals per sex per dose:

6

Mortality:

3.9 mg/L: 6/6 males died during the second half of exposure.

Clinical signs:

other: 3.9 mg/L: lethargy and nasal erythema were observed during the first two hours of exposure, nasal bleeding, ocular discharge and erythema, labored breathing, convulsion, collapse and death were noted during exposure period 2 to 5.5 h.

Gross pathology:

3.9 mg/L: lung hyperamia, fluid in pleural cavity.

Interpretation of results:

study cannot be used for classification

Reference 3

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1979-04-06 to 1979-04-20

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Remarks:

no raw data, particle size distribution not shown, particle size diameter selected to be less than 6.7 µm, frequency of observations not specified, temperature, humidity and pressure in the exposure chamber not reported, no necropsy performed

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

adopted 07 Sep 2009

Deviations:

yes

Remarks:

no raw data, particle size distribution not shown, particle size diameter selected to be less than 6.7 µm, frequency of observations not specified, temperature, humidity and pressure in the exposure chamber not reported, no necropsy performed

GLP compliance:

no

Test type:

traditional method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

inhalation: mist

Type of inhalation exposure:

whole body

Vehicle:

air

Remark on MMAD/GSD:

Droplet size determinations and counts were carried out by means of a Cascade Impactor. Particle size determination and particle counts in the atmosphere samples revealed that 95% of the particles had a diameter smaller than 6.7 µm.

Details on inhalation exposure:

Airflow: 1.5 m3/h The concentration used was the maximum attainable aerosol concentration. Whole body exposure. Analysis of test atmosphere every 45 minutes with an impinger. Receptor fluid: ethanol. GC Analysis of solution.

Analytical verification of test atmosphere concentrations:

yes

Remarks:

Analysis of test atmosphere every 45 minutes with an impinger.

Duration of exposure:

4 h

Concentrations:

4.33 mg/L air

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 weighing: at days 0, 3, 5, 7, 10, 12 and 14
- Necropsy of survivors performed: no

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 4.33 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

2/5 males and 0/5 females died on Day 3 and Day 12 of the observation period.

Clinical signs:

other: During the 4 h exposure period the rats were quiet, with their eyes tightly closed and showed nasal discharge. One male rat showed mouth breathing after 1 h of exposure. After exposure some rats had blood around their noses.

Body weight:

During the first three days of the observation period both males and females lost body weight. During the remainder part of the observation period 4/5 males and 5/5 females gained body weight.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

CLP: Acute Inhal. 4, H332 according to Regulation (EC) No. 1272/2008 based on mortality observed in males (2/5) at 4.33 mg/L air

Reference 4

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

(sampling time point for analytical determination not stated, mass median aerodynamic diameter not reported, dose levels too low, only males tested, 1h exposure only, 15 min post-observation period, no gross pathology performed)

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

- Principle of test: Groups of 5 male mice/group were exposed to inhalation of test item vapors for 1 h each.
- Short description of test conditions: Average test item concentrations were 12 and 1390 mg/m3 air.
- Parameters analysed / observed: mortality and clinical signs of the animals, oxygen and nitrogen content of the chamber.

GLP compliance:

no

Test type:

traditional method

Limit test:

no

Species:

mouse

Strain:

not specified

Sex:

male

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

air

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

1 h

Concentrations:

12 and 1390 mg/m3 air

No. of animals per sex per dose:

10

Control animals:

no

Details on study design:

- Duration of observation period following administration: The surviving animals were left in the chamber for further observation. Unfortunately, some condensation developed which allowed for accumulation of liquid test substance in the chamber. Approximately 15 min into the extended observation period, all of the animals had died.

Sex:

male

Dose descriptor:

LC50

Effect level:

> 12 mg/m³ air

Based on:

test mat.

Exp. duration:

1 h

Mortality:

12 mg/m3 air: no mortality occurred.
1390 mg/m3 air: 2/10 males died during exposure.

Clinical signs:

other: please refer to "Any other information on results incl. tables"

Clinical signs of toxicity:

12 mg/m3 air: restlessness and closed eyes during exposure

1390 mg/m3 air: restlessness and closed eyes during exposure, ataxia, gasping for breath and disturbance of motor coordiation after 37 min of exposure; sedation and labored breathing immediately after exposure

Interpretation of results:

study cannot be used for classification

Conclusions:

Based on the experimental findings, a LD50 value of > 12 mg/m3 air was determined. However, due to the significant methodological deficiencies (only 1 h exposure at too low dose levels, testing in males only etc.) the study cannot be used for classification.

Reference 5

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

adopted 07 Sep 2009

Deviations:

yes

Remarks:

particle size distribution was not determined, mass median aerodynamic diameter not given, only 2 dose levels but both close to limit concentrations, only one sampling time point for analytical determinations

GLP compliance:

yes

Test type:

traditional method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

air

Remark on MMAD/GSD:

No particle size analysis was performed during the study since the test material was determined during pretest to contain few, if any, particles.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

10.4 and 11.6 mg/L nominal concentration corresponding to 4.5 and 4.8 mg/L analytical concentration

No. of animals per sex per dose:

5

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: The placement of cages allowed only limited observation of animals during the exposure. No observations during exposure were made on the control group. All animals were thoroughly examined following exposure and gross signs of toxicity were recorded. During the observation period the animals were examined twice daily and gross signs of toxicity were recorded except on weekend when only mortality checks were done.
- Frequency of weighing: prior to exposure and post exposure on Days 2, 7 and 14
- Necropsy of survivors performed: The test animals were sacrificed at the end of the post-exposure period using chloroform, and a standard macroscopic examination of the external appearance and of tissues of the thoracic, abdominal and cranial cavities was conducted.

Key result

Sex:

male

Dose descriptor:

LC50

Effect level:

4.5 - 4.8 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Key result

Sex:

female

Dose descriptor:

LC50

Effect level:

> 4.8 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: No mortality was observed in females.

Mortality:

4.5 mg/L air: 1/5 males and 0/5 females died on Day 10 post-exposure.
4.8 mg/L air: 4/5 males and 0/5 females died between Day 1 and 9 post-exposure.

Clinical signs:

other: refer to "Any other information on results incl. tables"

Body weight:

4.5 and 4.8 mg/L: significant decreases in mean body weight in females on post exposure Day 2, significant decreases in mean body weight in males throughout the whole study period, body weights at study termination were below pre-exposure weights.

Gross pathology:

Necropsy observations included congested/red lungs, focus in lungs, focal corneal opacity, abnormal color of liver and congested/red turbinates. The majority of observations was noted in males. The findings were attributed to treatment.

Clinical signs of toxicity:

4.5 and 4.8 mg/L air: laboured breathing, gasping, clear nasal discharge, lacrimation and hypoactivity during exposure; labored breathing, gasping, clear nasal discharge, salivation and red discharge of eye immediately after exposure; gasping, red-brown perinasal encrustration, periorbital encrustration, urine stained fur and faeces stained fur during the post-exposure period. By Day 7 the only remaining signs of toxicity were red-brown perinasal encrustation and urine stained fur. By Day 14 only urine stained fur was observed.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

CLP: Acute Inhal. 4, H332 according to Regulation (EC) No. 1272/2008

Reference 6

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1987-11-02 to 1987-11-23

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:

adopted 07 Sep 2009

Deviations:

yes

Remarks:

no information on particle size distribution

GLP compliance:

yes

Test type:

traditional method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, Fuellinsdorf, Switzerland
- Age at study initiation: 9 weeks (males) and 11 weeks (females)
- Weight at study initiation: 262.4 - 289.1 g (males) and 207.0 - 234.8 g (females)
- Housing: in groups of 5 in Makrolon type 4 cages with standard softwood bedding
- Diet: pelleted standard Kliba 343 (Batch 83/87, from Kliba Klingentalmuehle, Kaiseraugst, Switzerland), ad libitum
- Water: community tap water, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Remark on MMAD/GSD:

The aerosol generation system was chosen to achieve the intended aerosol concentration with a mass median aerodynamic diameter (MMAD) of 3 µm or less. The particle size distribution is not shown in this study report. The MMAD itself and corresponding geometric standard deviation (GSD) were not determined.

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Flow past nose-only design.
- Method of holding animals in test chamber: The animals are confined separately in tubes which are positioned radially around the exposure chamber
- System of generating particulates/aerosols: The test article atmosphere was generated by a constant volume reservoir feeding a Hospitak No. 950 nebuliser following dissolution with water. Following the nebuliser, a dilution system was used to dilute the test article output of the nebuliser with clean air to concentration required for the study. Following aerolisation, the test article was discharged into the exposure chamber. The exposure aerosol enters the inlet at the top under a slight positive pressure and is distributed to the entrance of each of the feed tubes. The aerosol is then delivered through these tubes to the animal's nose and is then extracted away through a second concentric tube which is maintained under a negative pressure through aspiration at the outlet.
- Temperature, humidity and oxygen content were measured at the start of the exposure period: 22.5 °C, 16% humidity, 20.9% oxygen

TEST ATMOSPHERE
- Brief description of analytical method used: The test article was sampled from the exposure unit and passed through three bottles, each filled with 80 mL of ethyl acetate and cooled with dry ice. The ethyl acetate solutions were transferred into a 100 mL volumetric flask, the bottle was rinsed with ethyl acetate and the volume made up. The solutions were analysed by gas chromatography using a Carlo Erba HRGC 5300 apparatus.
- Samples taken from breathing zone: yes

VEHICLE
- Concentration of test material in air: Mean exposure concentration: 4.84 mg/L air. For details please refer to “Any other information on materials and methods incl. tables”.

TEST ATMOSPHERE
- Particle size distribution: The aerosol generation system was chosen to achieve the intended aerosol concentration with a mass median aerodynamic diameter of 3 µm or less, if technically possible.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

4.84 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: The animals were inspected for mortalility/viability once per hour during exposure, once after exposure on test Day 1 and twice daily thereafter. Symptoms were recorded once per hour during exposure (only grossly abnormal signs, due to the animals being in restraint tubes) and once daily thereafter.
- Frequency of weighing: At Day 1 (day of exposure), 8 and 15 of the test.
- Necropsy of survivors performed: yes
- Other examinations performed: The lungs from all animals were collected and kept in a neutral 4% formaldehyde solution.

Statistics:

Logit model could not be applied to the data, estimation without statistical analysis.

Key result

Sex:

male

Dose descriptor:

LC50

Effect level:

> 4.8 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: 2/5 males died

Key result

Sex:

female

Dose descriptor:

LC50

Effect level:

> 4.8 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: No mortality observed in females.

Mortality:

4.84 mg/L: 2/5 males and 0/5 females died (8 and 11 days after exposure)

Clinical signs:

other: Somnolence, hunched posture, piloerection and/or epistaxis in males and females after exposure, fully reversible within 24 h. In addition, rales were observed, which were fully reversible in males by Day 12 and fully reversible in females by Day 4.

Body weight:

The mean body weight gain shown over the study period was within the range expected for rats of this strain and age used in this type of study.

Gross pathology:

Necropsy of surviving animals revealed no abnormal findings. Reddening of the lung or spots on the lung lobes were observed in 2/2 dead males.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

CLP: Acute Inhal. 4, H332 according to Regulation (EC) No. 1272/2008 based on mortality observed in males (2/5) at 4.8 mg/L air

Reference 7

Endpoint:

acute toxicity: inhalation

Type of information:

other: publication (review article)

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

review article without experimental details, only LC50 values reported

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

GLP compliance:

no

Species:

mouse

Duration of exposure:

2 h

Concentrations:

5.4 and 11.1 mg/L (corresponding to 5400 and 11500 mg/m3 or 1254 and 2578 ppm)

Dose descriptor:

LC50

Effect level:

4.76 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

2 h

Remarks on result:

other: dose levels in mg/L air were calculated as follows: [dose level in ppm x molecular weight] / 24.2

Remarks:

(nominal concentration: 5.4 mg/L air)

Dose descriptor:

LC100

Effect level:

11.1 mg/L air (nominal)

Based on:

test mat.

Exp. duration:

2 h

Remarks on result:

other: dose levels in mg/L air were calculated as follows: [dose level in ppm x molecular weight] / 24.2

Interpretation of results:

study cannot be used for classification

Reference 8

Endpoint:

acute toxicity: inhalation

Type of information:

other: publication (review article)

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

review article without experimental details, only LC50 values reported

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

GLP compliance:

no

Species:

rat

Duration of exposure:

4 h

Concentrations:

5.82 mg/L (corresponding to 5820 mg/m3, 1339 ppm)

Dose descriptor:

LC50

Effect level:

4.8 mg/L air (analytical)

Exp. duration:

4 h

Remarks on result:

other: dose levels in mg/L air were calculated as follows: [dose level in ppm x molecular weight] / 24.2

Remarks:

(nominal concentration: 5.82 mg/L air)

Interpretation of results:

study cannot be used for classification

Reference 9

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

adopted 07 Sep 2009

Deviations:

yes

Remarks:

no experimental details, no information on analytical purity, no details on test animals, no information on frequency of observations, body weights not recorded, particle size distribution not determined, analytical concentrations not given for all doses

GLP compliance:

no

Test type:

traditional method

Limit test:

no

Species:

mouse

Strain:

other: Oncins France Strain 1 (OF1)

Sex:

not specified

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

1.2, 6.0, 12 and 24 mL/m3, corresponding to 1.24, 6.22, 12.42 and 24.86 mg/L

No. of animals per sex per dose:

8 / dose group (sex unspecified)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes

Sex:

not specified

Dose descriptor:

LC50

Effect level:

> 6.22 - < 12.43 mg/L air (nominal)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: corresponding to > 6 < 12 mL/mg3 air (nominal concentration), dose levels in mg/L air were calculated as follows: [dose level in ppm x molecular weight] / 24,200)

Mortality:

1.2 mL/m3 air (corresponding to 1.24 mg/L air): no mortality occurred
6.0 mL/m3 air (corresponding to 6.22 mg/L air): 2/8 animals died (after 2 h of exposure and on Day 1)
12 mL/m3 air (corresponding to 12.43 mg/L air ): 8/8 animals died (within 3 h during exposure)
24 mL/m3 air (corresponding to 24.86 mg/L air): 8/8 animals died (within the first hour of exposure)

Clinical signs:

other: Irritation of eyes and nasal mucosa was observed at the beginning of exposure.

Body weight:

No information on body weight available

Gross pathology:

There were no macroscopic lesions observed.

Interpretation of results:

study cannot be used for classification

Reference 10

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

adopted 07 Sep 2009

Deviations:

yes

Remarks:

no experimental details, no information on analytical purity, no details on test animals, no information on frequency of observations, body weights not recorded, particle size distribution not determined, analytical concentrations not given for all doses

GLP compliance:

no

Test type:

traditional method

Limit test:

no

Species:

rat

Strain:

other: Oncins France Strain A (OFA)

Sex:

not specified

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 200 - 300 g

Route of administration:

inhalation: vapour

Details on inhalation exposure:

The rats were placed into a 500 L inhalation chamber . They were exposed to the test item as vapour in air at a flow rate of 1m3/h. A saturation of the atmosphere with the test item vapour was noticed at the highest concentrations tested (e.g. 12 and 24 mL/m3)

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

1.2, 6.0, 12, 24 mL/m3, corresponding to 1.24, 6.22, 12.42 and 24.86 mg/L

No. of animals per sex per dose:

1.2 and 6.0 mL/m3 (corresponding to 1.24 and 6.22 mg/L air): 10 / dose group (sex unspecified)
12 and 24 mL/m3 (corresponding to 12.43 and 24.86 mg/L air): 8 / dose group (sex unspecified)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes

Sex:

not specified

Dose descriptor:

LC50

Effect level:

> 1.24 - < 24.86 mg/L air (nominal)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: corresponding to > 12 < 24 mL/m3 air (nominal), dose levels in mg/L air were calculated as follows: [dose level in ppm x molecular weight] / 24,200)

Mortality:

1.2 mL/m3 air: no mortality occurred
6.0 mL/m3 air: no mortality occurred
12 mL/m3 air: no mortality occurred
24 mL/m3 air: 8/8 animals died (6/8 died after 2-3 h of exposure, 2/8 died during post-exposure days 2 and 4)

Clinical signs:

other: Irritation of eyes and nasal mucosa was observed at the beginning of exposure.

Body weight:

No information on body weight available.

Gross pathology:

There were no macroscopic lesions observed.

Other findings:

The test atmosphere was saturated with test item vapour at ≥ 12 mL/m3 air, corresponding to ≥ 12.43 mg/L air. Below the saturation limit (at 1.2 and 6 mL/mg air, corresponding to 1.24 and 6.22 mg/L air) no mortality was expected.

Interpretation of results:

study cannot be used for classification

Reference 11

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

adopted 07 Sep 2009

Deviations:

yes

Remarks:

no particle size distribution given, no mass median aerodynamic diameter given

GLP compliance:

not specified

Test type:

traditional method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

air

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

1 - 6 h

Remarks on duration:

Static exposure: 1 and 6 h, dynamic exposure: 4 h

Concentrations:

Static exposure (1 hour): 408 ± 45 ppm (corresponding to 1.76 mg/L air)
Static exposure (6 hours): 328 ± 28 ppm (corresponding to 1.42 mg/L air)
Dynamic exposure (4 hours): 289 ± 3 ppm (corresponding to 1.24 mg/L air; nominal concentration: 295 ppm)

No. of animals per sex per dose:

Static exposure (1 and 6 hours): 5 females
Dynamic exposure (4 hours): 5/sex

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: All animals were observed for signs of toxicity on the day of exposure and for 14 days following exposure.
- Frequency of weighing: prior to exposure and on post-exposure Days 7 and 14.
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 289 ppm

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: corresponding to 1.2 mg/L air

Mortality:

Static exposure (1 hour, 408 ± 45 ppm): 5/5 females died within 3 days post-exposure, attributed to acrolein exposure (impurity).
Static exposure (6 hours, 328 ± 28 ppm): 5/5 females died during exposure, attributed to acrolein exposure (impurity).
Dynamic exposure (4 hours, 289 ± 3 ppm): no mortality was observed.

Clinical signs:

other: Static exposure (1 hour, 408 ± 45 ppm and 6 hours, 328 ± 28 ppm): respiratory difficulties, periocular, perinasal and perioral wetness. Dynamic exposure (4 hours, 289 ± 3 ppm): no clinical signs were observed.

Body weight:

Dynamic exposure (4 hours, 289 ± 3 ppm): depression of body weight during the first week post-exposure.

Gross pathology:

Static exposure (1 hour, 408 ± 45 ppm and 6 hours, 328 ± 28 ppm): discoloration of the lungs and liver, clear fluid in the trachea and pleural cavity, eye opacity.
Dynamic exposure (4 hours, 289 ± 3 ppm): no macroscopic abnormalities were observed.

Interpretation of results:

study cannot be used for classification

Conclusions:

There was no mortality after acute inhalation in male and female rats at concentrations up to 1.2 mg/L air. The dose levels were below the recommended limit dose of current guidelines, therefore no conclusion on classification can be drawn.

Reference 12

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

mass median aerodynamic diameter not given

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

- Principle of test: Groups of male and/or female rats were exposed to a saturated vapor atmosphere of the test item for different times of exposure. The test atmosphere was generated statically or dynamically.
- Short description of test conditions:
1. Static exposure: Whole body exposure
120 - 250 g of liquid test substance were placed in an open tray at the top of a sealed 120 or 250 L stainless steel and glass exposure chamber. The atmosphere was allowed to equilibrate for a minimum of 6 h. The animals were then introduced into the chamber through a gasketted door. The test item atmosphere was generated as supplied or from test item which was previously sparged with nitrogen in an attempt to remove volatile impurities, notably acrolein.
The animals were exposed for 24, 40, 49, 60, 80, 97, 100, 195, 240, 360 and 380 min (refer to Table no. 1 under "Any other information on materials and methods incl. tables").
- Parameters analysed / observed: clinical signs, mortality, gross necropsy, histopathological examination of lungs (only from animals exposed for 60 and 240 min); analytical determinations of the test item in the exposure chamber atmosphere.

2. Dynamic exposure: Whole body and nose-only exposure
The atmosphere was generated by metering liquid test substance from a piston pump into a heated evaporator maintained at the lowest temperature (91°C) sufficient to vaporise the liquid.
Whole body exposure was performed in a 900 L stainless steel glass and steel chamber, in which a countercurrent flow of air was passed through the vaporiser and vapor containing air was passed through the chamber.
For nose-only exposure, test item containing air was introduced at the top of the chamber, flowed past the nose-only ports and exhausted at the bottom of the chamber. The atmosphere was sampled at 6 min intervals to measure the test item and acrolein concentration (refer to Table no. 2 under "Any other information on materials and methods incl. tables").
- Parameters analysed / observed: clinical signs, mortality, body weights, gross necropsy, histopathological examination of lungs (only from nose-only exposed animals); analytical determinations of the test item in the exposure chamber atmosphere.

GLP compliance:

not specified

Test type:

traditional method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

inhalation: vapour

Type of inhalation exposure:

other: whole body and nose only

Vehicle:

air

Remarks:

in some cases sparged with liquid nitrogen, refer to table no. 1

Analytical verification of test atmosphere concentrations:

yes

Remarks:

for 60, 240, 360 and 380 min exposure only

Duration of exposure:

24 - 380 min

Concentrations:

60 min static exposure: 408 ± 4.5, 733 ± 22 and 935 ppm
240 min static exposure: 957 ± 220 ppm
360 min static exposure: 261 ± 20, 270 ± 51 and 328 ± 4.5 ppm
380 min static exposure: 261 ± 2 ppm
(Please refer to Tables no. 1 and 2 under "Any other information on materials and methods incl. tables".)

No. of animals per sex per dose:

5/sex (except for 935 ppm: 3 male, 7 female)

Control animals:

no

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 289 ppm

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: whole body exposure

Remarks:

corresponding to 1.19 ± 0.0081 mg/L air

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 306 ppm

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: nose-only exposure

Remarks:

corresponding to 1.32 ± 0.344 mg/L air

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 320 ppm

Based on:

test mat.

Exp. duration:

1 h

Remarks on result:

other: nose only exposure

Remarks:

corresponding to 1.38 mg/L air

Mortality:

Static exposure (whole body exposure for 24 - 380 min at 261 - 957 ppm): all animal of all concentrations and all exposure durations died (refer to Table no. 3 under "Any other information on results incl. tables").
Dynamic exposure (4 h exposure, whole body at 277 and 289 ppm): no mortality occurred.
Dynamic exposure (1 h exposure, nose-only at 320 ppm): no mortality occurred.
Dynamic exposure (4h exposure, whole body at 306 ppm): 1/5 males and 0/5 females died on Day 4 post exposure.

Clinical signs:

other: Please refer to "Any other information on results incl. tables"

Body weight:

Static exposure (whole body exposure for 24 - 380 min at 261 - 957 ppm): body weights were not recorded
Dynamic exposure (4 h whole body exposure at 277 and 289 ppm): reduced body weight gain during the whole study period
Dynamic exposure (1 and 4 h nose only exposure at 320 and 306 ppm): reduced body weight gain during the first week

Gross pathology:

Static exposure (whole body exposure for 24 - 380 min at 261 - 957 ppm): dark red lungs, clear fluid in the trachea and thoracic cavity, livers with dark purple appearance, epithelium of the trachea and major bronchi were detached and there were areas of eosinophilic fibrillar material along the walls.
Dynamic exposure (4 h whole body exposure at 277 and 289 ppm, 1 and 4 h nose only exposure at 320 and 306 ppm): no abnormalities observed.

Other findings:

Histopathology of lungs:
Static exposure (whole body exposure for 24 - 380 min at 261 - 957 ppm): mild to marked congestion, edema, scattered areas of alvolar haemorrhage.
Dynamic exposure (4 h whole body exposure at 277 and 289 ppm, 1 and 4 h nose only exposure at 320 and 306 ppm): no abnormalities observed.

Clinical signs of toxicity:

Static exposure (whole body exposure for 24 - 380 min at 261 - 957 ppm): lacrimation, nasal discharge, mouth and laboured breathing during exposure, wheezing rales and gasping breathing post-exposure.

Dynamic exposure (4 h whole body exposure at 277 and 289 ppm, 1 and 4 h nose only exposure at 320 and 306 ppm): slight lacrimation, bleopharospasm, occasional slight uniform salivation.

Table 3: Time to death in animals exposed under static conditions

Exposure time [min]	Sex	No. of animals/sex	Atmosphere sparged	Concentration [ppm] mean ± SD	Time of death
24	female	5	No	NM	1-9 days post exposure
40	male	5	No	NM	1-2 days post exposure
49	female	5	No	NM	1-2 days post exposure
60	female	5	No	408 ± 4.5	4 h to 1 day post exposure
60	male/female	3 male, 7 female	No	935	Day 1 post exposure
60	male/female	5	Yes	733 ± 22	Day 1 post exposure
80	male	5	No	NM	1 during exposure, 4 on Day 4 post exposure
97	female	5	No	NM	3 during exposure, 2 on Day 1 post exposure
100	male	5	No	NM	during exposure
195	female	5	No	NM	during exposure
240	male/female	5	No	957 ± 220	during exposure
360	female	5	No	261 ± 20	during exposure
360	female	5	Yes	328 ± 4.5	during exposure
360	male/female	5	Yes	270 ± 51	during exposure
380	male	5	No	261 ± 2.4	during exposure

Interpretation of results:

study cannot be used for classification

Conclusions:

The LD50 values in male and female rats were > 1.19 ± 0.0081 mg/L air (for 4 h whole body exposure) and > 1.32 ± 0.344 mg/L air (for 4 h nose-only exposure). The dose levels were below the recommended limit dose of current guidelines, therefore no conclusion on classification can be drawn.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

4.8 mg/m³

Quality of whole database:

The available information comprises adequate and reliable studies, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.5, of Regulation (EC) No. 1907/2006.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

(no information on analytical purity, no experimental details, only one rabbits per dose level, no details given on animals and environmental conditions, duration of exposure and duration of the observation period not specified)

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Acute dermal toxicity was determined by skin absorption in rabbits.
- Short description of test conditions: Undiluted test substance was applied to the closely clipped, intact skin of albino rabbits. Treated areas were covered with plastic shields to reduce evaporation.
- Parameters analysed / observed: Mortality, clinical signs, autopsy of decedent animals

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

other: albino

Sex:

male/female

Type of coverage:

occlusive

Vehicle:

not specified

Doses:

1000, 1250, 1750, 2250, 2500 and 3100 mg/kg bw

No. of animals per sex per dose:

1

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: several days
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 250 mg/kg bw

Based on:

test mat.

Mortality:

1000 mg/kg bw: no mortality occurred.
1250 mg/kg bw: no mortality occurred.
1750 mg/kg bw: no mortality occurred.
2500 mg/kg bw: 1/1 males died within 8 h post application.
3100 mg/kg bw: 1/1 males died over night post application.

Clinical signs:

1000 and 1250 mg/kg bw: no clinical signs of toxicity.
1750, 2500 and 3100 mg/kg bw: weakness was observed within 30 min, nervousness followed by lethargy and irregular breathing. Surviving animals recovered after several days.

Gross pathology:

1000, 1250 and 1750 mg/kg bw: no necropsy performed.
2500 and 3100 mg/kg bw: necropsy revealed no abnormal findings, except for discoloration at the side of application.

Interpretation of results:

study cannot be used for classification

Conclusions:

Based on the experimental findings, a LD50 value of > 2250 mg/kg bw was determined. However, only one animal was used per dose level and no information is available on duration of exposure or the observation period. Due to the significant methodological deficiencies the study cannot be used for classification.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted 09 Oct. 2017

Deviations:

yes

Remarks:

no experimental details given, no body weight and no clinical signs recorded, no scoring according to Draize criteria, no data on analytical purity, no details on animals and environmental conditions, occlusive dressing

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: OFA

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 250 g (males) and 200 g (females)

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Duration of exposure:

24 h

Doses:

0.25, 0.5, 1, 2 mL/kg bw corresponding to 259, 518, 1036 and 2072 mg/kg bw

No. of animals per sex per dose:

0.25, 0.5 and 1 mL/kg bw (corresponding to 259, 518, 1036 mg/kg bw): 5
2 mL/kg bw (corresponding to 2072 mg/kg bw): 10

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

673 mg/kg bw

Based on:

test mat.

Remarks on result:

other: corresponding to 0.65 mL/kg bw, dosel levels in mg/kg bw calculated with a density of 1036 mg/mL

Mortality:

0.25 mL/kg bw corresponding to 259 mg/kg bw: 0/5 males and 1/5 females died within 24 h.
0.5 mL/kg bw corresponding to 518 mg/kg bw: 0/5 males and 4/5 females died within 24 h.
1 mL/kg bw corresponding to 1036 mg/kg bw: 4/5 males and 2/5 females died within 24 h.
2 mL/kg bw corresponding to 2072 mg/kg bw: 10/10 males and 10/10 females died within 2 h.

Clinical signs:

Severe oedema was observed after 24 h. Scab formation was observed that detached from the skin between 4 to 15 days post application.

Body weight:

No information available

Gross pathology:

No significant macroscopic abnormalities were observed.

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

CLP: Acute Dermal 3, H311 according to Regulation (EC) No. 1272/2008

Reference 3

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

only short summary, only 1 animal per dose

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Male and female New Zealand rabbits received a dermal application of the test substance. One animal was used per dose.
- Short description of test conditions: No experimental details given. Doses ranged from 1.26 to 7.94 mg/kg bw.
- Parameters analysed / observed: Mortality, clinical signs, gross necropsy.

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Vehicle:

unchanged (no vehicle)

Duration of exposure:

24 h

Doses:

1260, 2000, 3160, 5010 and 7940 mg/kg bw

No. of animals per sex per dose:

1

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

1260 mg/kg bw: no mortality occurred.
2000 mg/kg bw: no mortality occurred.
3160 mg/kg bw: 1/1 males died within 2 days.
5010 mg/kg bw: 1/1 females died within 24 h.
7940 mg/kg bw: 1/1 males died within 24 h.

Clinical signs:

Increasing weakness, diarrhea, collaps; reduced appetite and activity were noted 3-6 days after exposure in surviving rabbits.

Gross pathology:

Hemorrhaging areas of the lungs, liver and kidney discoloration and gastrointestinal inflammation.

Interpretation of results:

study cannot be used for classification

Reference 4

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Remarks:

no details on animals and environmental conditions, body weights not recorded, no scoring according to Draize, no details on analytical purity

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted 09 Oct 2017

Deviations:

yes

Remarks:

no details on animals and environmental conditions, body weights not recorded, no scoring according to Draize, no details on analytical purity, occlusive dressing, body weight range of males too large

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Central Institute for the Breeding of Laboratory Animals TNO, Zeist, The Netherlands
- Weight at study initiation: 154 - 222 g (males) and 110 - 154 g (females)
- Housing: in groups of 5 in screen-bottomed stainless-steel cages
- Diet: Stock diet, ad libitum
- Water: tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 1

Type of coverage:

occlusive

Vehicle:

water

Duration of exposure:

24 h

Doses:

1.73, 2.00, 2.49 and 3.00 mL/kg bw, corresponding to 1792, 2072, 2580 and 3108 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: during the 24 h application and daily throughout the post-treatment weeks
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

2 621 mg/kg bw

Based on:

test mat.

95% CL:

2 352 - 2 922

Remarks on result:

other: dosel levels in mg/kg bw calculated with a density of 1036 mg/mL

Mortality:

1.73 mL/kg bw, corresponding to 1792 mg/kg bw: 2/5 males and 0/5 females died within 1-2 h post administration.
2.00 mL/kg bw, corresponding to 2072 mg/kg bw: 2/5 males and 2/5 females died within 1-2 h post administration.
2.49 mL/kg bw, corresponding to 2580 mg/kg bw: 4/5 males and 01/5 females died within 1-2 h post administration.
3.00 mL/kg bw, corresponding to 3108 mg/kg bw: 5/5 males and 5/5 females died within 1-2 h post administration.

Clinical signs:

Discomfortable behaviour, aggressiveness, aimless runnning and convulsive movements were observed a few minutes after treatment. Later on signs of asphyxation and coma were frequently observed. After 24 hours the surviving animals appeared healthy again. There were no signs of erythema or oedema observed.

Gross pathology:

Necropsy revealed no treatment-related findings

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008

Reference 5

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted 09 Oct 2017

Deviations:

yes

Remarks:

dose level too low, no details on animals and environmental conditions, no information on analytical purity, no body weight recorded, no scoring according to Draize criteria, observation period of 24 h only, no necropsy performed

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.52 - 3.25 kg
- Housing: individually
- Diet: standard laboratory diet, ad libitum
- Water: tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): approx. 18

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

Application of a 10% aqueous solution

Duration of exposure:

24 h

Doses:

200 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 24 h
- Necropsy of survivors performed: no

Sex:

male/female

Dose descriptor:

other: NOED

Effect level:

> 200 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed.

Clinical signs:

Treatment-related skin effects consisted of a slight to moderate erythema and slight to severe edema. There were no abnormalities with respect to condition or behaviour.

Interpretation of results:

study cannot be used for classification

Conclusions:

Based on the experimental findings the no-observed-effect-dose was > 200 mg/kg bw. However, the dose level of 200 mg/kg bw chosen for the present study was too low and the observation period of 24 hours too short to draw any conclusion.

Reference 6

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1985-07-22 to 1982-10-09

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

EPA OPP 81-2 (Acute Dermal Toxicity)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted 09 Oct 2017

Deviations:

yes

Remarks:

no information on analytical purity, occlussive dressing, frequency of observations less than required according to guideline, no scoring according to Draize criteria

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hazleton-Dutchland Laboratory Animals Denver, Pennsylvania, USA
- Females nulliparous and non-pregnant: yes
- Age at study initiation: at least 8 weeks
- Weight at study initiation: 2.2 - 3.0 kg (males) and 2.4 - 3.1 kg (females)
- Housing: Individually in suspended, stainless steel cages with wire mesh bottoms
- Diet: Lab Rabbit Chow HF (Purina), ad libitum
- Water: municipal tap water, supplied by an automatic watering system, ad libitum
- Acclimation period: 16 - 38 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15 - 21
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 22 Jul 1985 To: 09 Oct 1985

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Duration of exposure:

24 h

Doses:

1000, 1500 and 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: approximately 1, 2 and 4 h after dosing and daily thereafter
- Frequency of weighing: at the time of clipping, before dosing and on study Days 7 and 14
- Necropsy of survivors performed: yes

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

1 600 mg/kg bw

Based on:

test mat.

95% CL:

1 271 - 1 929

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

1 500 mg/kg bw

Based on:

test mat.

95% CL:

1 106 - 1 894

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

1 650 mg/kg bw

Based on:

test mat.

95% CL:

1 409 - 1 891

Mortality:

1000 mg/kg bw: 1/5 males and 0/5 females died on Day 3.
1500 mg/kg bw: 0/5 males and 3/5 females died within 6 h to Day 4.
2000 mg/kg bw: 4/5 males and 4/5 females died within 4 - 24 h.

Clinical signs:

1000 mg/kg bw: ataxia in 2/5 males and 4/5 females at 4 hours, hyperactivity (in 5/5 males and 5/5 females) at 4 hours after patch removal; decreased food consumption for one or more days after dosing in 4/5 males and 3/5 females; soft mucoidal stool (0/5 males and 1/5 females), fecal staining (1/5 males and 0/5 females) and abdominal bloating during the second week (0/5 males and 1/5 females), indicative of intestinal disease, in line with pathological findings but not treatment-related.
1500 mg/kg bw: hyperpnea (0/5 males and 1/5 females), dyspnea (0/5 males and 1/5 females), irregular breathing (4/5 males and 1/5 females), occasional hypopnea (0/5 males and 1/5 females) were observed during the first 4 hours after patch removal, hyperactivity (3/5 males and 2/5 females) and prostration (1/5 males and 1/5 females) up to 24 hours after patch removal, swollen scrotum in 1/5 males on Days 2 and 3.
2000 mg/kg bw: hyperpnea (0/5 males and 1/5 females), dyspnea (4/5 males and 3/5 females), irregular breathing (0/5 males and 1/5 females), occasional hypopnea (1/5 males and 1/5 females) were observed during the first 4 hours after patch removal, hyperactivity (5/5 males and 5/5 females) and prostration (4/5 males and 3/5 females) up to 24 hours after patch removal.

Body weight:

All animals exhibited body weight loss at Day 7 (- 3.6 – 7.2% at 1000 and 1500 mg/kg bw in males and females, -12.2% in males and -16.2% in females at 2000 mg/kg bw). In addition, most animals had slight weight gains or no weight change between Days 7 and 14 (-3.7% to -7.5% at 1000 mg/kg bw in males and females, up to -3.7% at 1500 mg/kg bw in males and females and -9% at 2000 mg/kg bw in males and females). 1/5 females at 1000 mg/kg bw had large weight loss (-18.7%) at Day 14. Findings on body weight development were in line with decreased food consumption (please refer to “Other findings”).

Gross pathology:

1000 mg/kg bw: dermal lesions in all animals, treatment-related; intestinal disease (mucoid enteritis) in 1/5 males and 1/5 females, not treatment-related.
1500 mg/kg bw: dermal lesions in all animals, treatment-related; enlarged heart in 1/5 females, not treatment-related.
2000 mg/kg bw: dermal lesions in all animals, treatment-related.

Other findings:

Food consumption:
Food consumption was decreased in 4/5 males and 3/5 females at 1000 mg/kg bw during the first 2 days of the study and remained decreased in 1/5 males and 2/5 females until study termination. At 1500 mg/kg bw, food consumption was decreased in 3/5 males and 2/5 females but was comparable to normal levels by study Day 4. At 2000 mg/kg bw, food consumption was decreased in the single surviving female on study Days 7 – 11.

Local signs of toxicity:
All animals surviving at 24 h exhibited severe dermal effects at the dose site (necrosis followed by eschar formation and/or exfoliation of the eschar tissue) which persisited throughout the study.

The effect levels described in the result table above were calculated including the mortality of a single male rabbit at 1000 mg/kg bw. When excluding the mortality of one male at 1000 mg/kg bw, the LD50 value for males was 1800 mg/kg bw (95% confidence interval: 1552 - 2048) and the LD50 value for males and females combined was 1700 mg/kg bw (95% confidence interval: 1467 - 1933).

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

CLP: Acute Dermal 4, H312 according to Regulation (EC) No. 1272/2008

Reference 7

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted 09 Oct 2017

Deviations:

yes

Remarks:

occlusive dressing, no stepwise dosing

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

yes

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: clipped dorsal trunk
- Type of wrap if used: The test material was applied to the test site and the exposed skin was covered with gauze and impervious sheeting. The materials were held in place using an elastic bandage around the trunk.

Duration of exposure:

24 h

Doses:

0.25, 0.5, 1.0 mL/kg bw (males), corresponding to 259, 518 and 1036 mg/kg bw and 0.5, 1.0 mL/kg bw (females), corresponding to 518 and 1036 mg/kg bw

No. of animals per sex per dose:

4

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: during exposure and daily thereafter
- Frequency of weighing: before dosing, at 7 and 14 days post dosing
- Necropsy of survivors performed: yes

Sex:

male

Dose descriptor:

LD50

Effect level:

736 mg/kg bw

Based on:

test mat.

95% CL:

446 - 1 191

Remarks on result:

other: dosel levels in mg/kg bw calculated with a density of 1036 mg/mL

Sex:

female

Dose descriptor:

LD50

Effect level:

818 mg/kg bw

Based on:

test mat.

95% CL:

508 - 1 347

Remarks on result:

other: dosel levels in mg/kg bw calculated with a density of 1036 mg/mL

Mortality:

0.25 mL/kg bw (corresponding to 259 mg/kg bw): 1/4 males died on Day 9.
0.5 mL/kg bw (corresponding to 518 mg/kg bw): no mortality occurred.
1.0 mL/kg bw (corresponding to 1036 mg/kg bw): 4/4 males and 3/4 females died (males: death within 3 h - 5 days; females: death within 1 day).

Clinical signs:

Local signs: Marked erythema, edema and necrosis in all animals of all dose groups, which persisted 14 days. Desquamation and fissuring were seen at 7 days in females at 1 mL/kg bw (1036 mg/kg bw).
Clinical signs: Salivation, sluggishness, and unsteady gait. Survivors recovered from these effects by 2 days.

Body weight:

Body weight gain was unaffected by treatment.

Gross pathology:

The main necropsy finding was light to dark red mottling of the lungs. Sacrificed survivors had no gross pathological changes observed.

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

CLP: Acute Dermal 3, H311 according to Regulation (EC) No. 1272/2008

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

736 mg/kg bw

Quality of whole database:

The available information comprises adequate and reliable studies, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.5, of Regulation (EC) No. 1907/2006.

Additional information

Acute oral toxicity:

Several studies assessing the acute oral toxicity of MMP are available.

In a key study performed similar to OECD guideline 401 and compliant with GLP, the test item was administered to male and female Sprague-Dawley-derived CDR rats (86-0243-FKT). The test item was administered unchanged by oral gavage as a single dose. Groups of 5 males per dose received doses of 250, 350, 500, 600 and 700 mg/kg bw, and groups of 5 females per dose received doses of 700, 850, 1000, 1200 and 1700 mg/kg bw. Following administration, the animals were observed for a period of 14 days for clinical signs of toxicity, including reduced food consumption, and mortality. The body weight was recorded weekly. At study termination, all decedent and surviving animals were subjected to gross necropsy.

Mortality was observed in males at ≥ 500 mg/kg bw and in females at ≥700 mg/kg bw; most deaths occurred within the first 4 h post-dosing up to 2 days after administration. Clinical signs of toxicity were noted at ≥ 250 mg/kg bw and included ocular and nasal discharge, hypopnea, dyspnoea, urinary staining and hyperactivity after dosing. The adverse effects were noted in up to 4/5 males at 250 mg/kg bw, in up to 4/5 males at 350 mg/kg bw, up to 4/5 males at 500 mg/kg bw, in up to 4/5 males at 600 mg/kg bw, in up to 5/5 males and 4/5 females at 700 mg/kg bw, in up to 3/5 females at 800 mg/kg bw, in up to 4/5 females at 1000 mg/kg bw, in up to 3/5 females at 1200 and in up to 4/5 females at 1700 mg/kg bw. In surviving animals, the adverse effects were fully reversible by study Day 3.

 At ≥ 500 mg/kg bw ataxia, dry and wet rales, prostration and partial closing of eyes was noted. The observations were made in 2/5 males at 500 mg/kg bw, in 2/5 males at 600 mg/kg bw, in up to 2/5 males at 600 mg/kg bw, in up to 5/5 females at 700 mg/kg bw, in 1/5 females at 850 mg/kg bw, in 1/5 females at 1000, in 3/5 females at 1200 and in up to 4/5 females at 1700 mg/kg bw. All signs were fully reversible in surviving animals by Days 4-5. Most animals had decreased food consumption beginning on the day after dosing, but body weight development was not affected by treatment.

At the post mortem examination discolouration of the lungs was noted in nearly all the male and female animals and red foci of the lungs was reported in the majority of male and female rats in all dose groups. In the animals sacrificed at study termination, these observations were attributed by the study author to the CO₂used to sacrifice the animals. However, it is unclear why animals that died during the observation period showed similar effects in the lungs. In all the males found dead during the observation period (≥ 500 mg/kg bw/day), red intestinal walls and red fluid in the intestine was observed. In a few individuals red stomach walls were also noted. In 1-2 females in each dose group from 850 mg/kg bw/day red intestinal walls and red fluid in the intestine was observed. These findings are considered to be toxicologically relevant. Test material was also found in the stomach of 4/5 and 4/5 females in the 1200 and 1700 mg/kg bw/day groups, respectively, and 2/5 males in the 700 mg/kg bw/day group.

 Reddened or swollen uterus and ovaries were observed in 3/5, 2/5, 3/5 and 2/5 females in the 700, 850, 1000, 1000, 1200 and 1700 mg/kg bw/day groups, respectively. The toxicological relevance of this finding is unclear.

Under the conditions of the test, the acute oral LD50 value was 490 mg/kg bw in males and 1050 mg/kg bw in females.

 

In the 8 supporting studies with reliability 2, 3, or 4, the acute oral LD50 values varied substantially between the studies. In the studies given reliability 2, the acute oral LD₅₀values for male rats fell in the range of 755 to 5926 mg/kg bw, and for female rats in the range of 755 to 6579 mg/kg bw. An LD₅₀value of 1620 mg/kg bw was given for mice in a study with reliability 4. 

Acute inhalation toxicity:

The acute inhalation toxicity of MMP was investigated in a total of 12 studies with reliability 2, 3 or 4.

In the key study conducted according to OECD guideline 403 and in compliance with GLP, 5 male and 5 female Wistar rats were exposed for 4 h via nose-only inhalation (88-0045-DGT). The test item was administered as an aerosol at a mean target concentration of 4.84 mg/L air. The aerosol generation system was chosen to achieve the intended aerosol concentration with a mass median aerodynamic diameter (MMAD) of 3 µm or less. After exposure, the animals were observed for a period of 14 days. Clinical signs of toxicity and mortality were recorded once per hour during exposure and twice daily during the observation period. Body weights were recorded on study Days 1, 8 and 15. At study termination all animals underwent macroscopic examination.

Mortality was observed in 2/5 males only and occurred on Days 8 and 11. Somnolence, hunched posture, piloerection and/or epistaxis were noted in males and females after exposure but were fully reversible within 24 h. In addition, rales were observed, which were fully reversible in males by Day 12 and fully reversible in females by Day 4. The mean body weight gain shown over the study period was within the range expected for rats of this strain and age used in this type of study. The male rats that died on Day 11 had lost body weight on Day 8, leading to a lower mean body weight on the Day 8 compared with Day 1. Macroscopic examination at scheduled necropsy showed no treatment-related findings in surviving animals. Reddening of the lung or spots on the lung lobes were observed in 2/2 dead males. The observations were attributed by the study author to the irritating potential of acrolein, which is an impurity in the test material.

Under the conditions of the test, the acute inhalation LC₅₀value was > 4.84 mg/L air for male and female rats.

 

In the 11 supporting studies with reliability 2, 3, or 4, the acute inhalation LC50 values were fairly consistent between the studies. For the studies considered reliability 2, acute inhalation LC₅₀values in the range of 4.33 to 5.8 mg/L air for male and female rats and an LC₅₀value of 5.4 mg/L air for mice were given. All studies performed use test material containing up to 0.14% acrolein as an impurity. 

In the OECD SIDS Initial Assessment Report (SIAM 17; Arona Nov. 2003; 3-(Methylthio) propionaldehyde) the findings on acute inhalative exposure are commented as follows: "It is unclear whether the observed local symptoms in the inhalation studies are attributable to 3-(methylthio) propionaldehyde or to acrolein which is contained in the test material at a concentration of up to 0.3 % in the materials tested. The irritation threshold for acrolein in rat nasal epithelium was approximately 0.25 ppm (0.6 mg/m3). As the developmental toxicity study has shown, the actual concentration of acrolein in the inhalation chamber can be much higher i.e. about 2% of the concentration of 3-(methylthio) propionaldehyde and it is possible that acrolein can be enriched in the vapor phase under the test conditions. However, at concentrations up to 216 mg/m3 (50 ppm) no respiratory irritation was detected in a study with repeated exposure to acrolein-free 3-(methylthio) propionaldehyde. MMP has a vapor pressure at 27 degree Centigrade of 1 mbar (1 hPa) leading to a vapor saturation concentration (VSC) of approx. 4.3 mg/L. The LC50 of 4.5 – 4.8 mg/L /4hrs determined in the inhalation study conducted by Monsanto (1986) therefore equals approximately the VSC and requires that MMP under these conditions has to be classified as an aerosol according to CLP.”

 

 Acute dermal toxicity:

The acute dermal toxicity of MMP was investigated in seven studies, of which one study was selected as key study.

The key study was performed similar to OECD guideline 402 (Ballantyne 2000), GLP status not specified. The test item was applied unchanged (no vehicle) to the clipped dorsal trunk of 4 male and 4 female rabbits for 24 h under occlusive conditions. Males were exposed to 0.25, 0.5 and 1.0 mL/kg bw, corresponding to 259, 518 and 1036 mg/kg bw; and females were exposed to 0.5 and 1.0 mL/kg bw, corresponding to 518 and 1036 mg/kg bw.

During exposure and during the 14-days observation period thereafter, the animals were inspected for clinical signs and mortality. Body weight was recorded at weekly intervals, and at study termination all surviving animals were examined for gross pathological abnormalities. The skin irritation effects (erythema and oedema) at the test site were recorded 1 h, and 1, 2, 3, 7, 10, 14 and 17 days after patch removal.

Mortality occurred in 1/4 males at 0.25 mL/kg bw (259 mg/kg bw) on study Day 9, in 4/4 males at 1.0 mL/kg bw (1036 mg/kg bw) between 3 h post-dosing and study Day 5 ;and in 3/4 females at 1.0 mL/kg bw (1036 mg/kg bw) within one day after dosing. Clinical signs of toxicity included salivation, sluggishness, and unsteady gait. Survivors recovered from these effects within 2 days. In addition, all animals of all dose groups showed marked erythema, edema and necrosis, which persisted for 14 days. Desquamation and fissuring were observed one week after dosing in females at 1 mL/kg bw (1036 mg/kg bw). Body weight gain was unaffected by treatment. The main necropsy finding was light to dark red mottling of the lungs in decedent animals. Gross macroscopical analysis of surviving animals revealed no abnormalities.

The acute dermal LD₅₀value was 736 mg/kg bw in male rabbits and 818 mg/kg bw in female rabbits.

 

Within the six supporting studies with reliability 2, 3 or 4 several treatment-related clinical signs of toxicity were noted, which comprised hyperpnea, dyspnea, irregular breathing, occasional hypopnea during exposure and after patch removal, as well as hyperactivity, discomfortable behavior and prostration following exposure. In addition, decreased food consumption and body weight loss was observed. Treatment-related skin effects consisted of a slight to moderate erythema and slight to severe edema. In all studies, dermal effects were observed at the testing site. Except for the dermal lesions, no significant macroscopic abnormalities were observed. The acute dermal LD₅₀ values ranged from 1106 – 1929 mg/kg bw in rabbits and from 673 – 2621 mg/kg bw in rats.

Justification for classification or non-classification

The available data on acute oral toxicity meet the criteria for classification as Acute Oral 4 (H302) according to Regulation (EC) No. 1272/2008 (CLP) and according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations.

The available data on acute inhalation toxicity with the aerosol meet the criteria for classification as Acute Inhal 4 (H332) according to Regulation (EC) No. 1272/2008 (CLP) and according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations.

The available data on acute dermal toxicity meet the criteria for classification as Acute Dermal 3 (H311) according to Regulation (EC) No. 1272/2008 (CLP) and according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations.