2-methylbutyric acid

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Cross-reference

Cross-reference 1

Reason / purpose:

reference to same study

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 200 - 300 g
- Fasting period before study: overnight
- Diet: standard lab diet ad libitum
- Water: ad libitum

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 4 mL/kg bw

Doses:

males: 1.0, 2.0, 4.0 mL/kg bw
females: 0.25, 0.5, 1.0, 2.0, 4.0 mL/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: on days 0 (prior to treatment), 7, and 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

LD50 value was calculated using the moving average method (Thompson, 1947; Weil, 1983)

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

1 750 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Using a density of 0.936 kg/L, the calculated LD50 of 1.87 mL/kg bw is equivalent to 1750 mg/kg bw

Mortality:

Most deaths occurred at 1.5 to 2 days. However, three females died at 10 to 14 days. Survivors recovered at one to three days.

Males: 0/5, 3/5 and 5/5 of the low, middle and high dose group died
Females: 0/5, 0/5, 1/5, 2/5, 5/5 of the 0.25, 0.5, 1, 2, 4 mL/kg bw dose group died, respectively

Clinical signs:

Signs of toxicity included sluggishness, lacrimation, unkempt fur, kyphosis (in 2), prostration, a moribund appearance, diarrhea, red to brown discharge on perioral, perinasal and periurogenital fur.

Body weight:

Males: body weight gain at 1.0 mL/kg bw; bodyw eight reduced at 2.0 mL/kg bw
Females: no body weight gain at all dose levels

Gross pathology:

Dead animals: stomach abnormalities (discoloration, brown liquid or gas, gastric collapse); discoloure dlungs; brown to dark red livers, intstines, and kidneys.
Survivors: small stomachs and spleens; rough stomach surfaces, adhesion of the stomach to liver or spleen, enlarged and mottled red kidneys.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The LD50 was 1750 mg/kg bw in male and female rats. Necropsy findings in survivors revealed irritation/corrosion of the stomach and intestines.

Executive summary:

Oral toxicity was investigated in a study similar to OECD TG 401. Male and female rats (n=5 per sex per dose) were exposed against different doses of test substance up to 4 mL/kg bw (3744 mg/kg bw by calculation on basis of substance density of 0.936 kg/L). Exposed animals showed signs of toxicity including sluggishness, lacrimation, unkempt fur, kyphosis, prostration, a moribund appearance, diarrhea, red to brown discharge on perioral, perinasal and periurogenital fur. Female rats of all dose groups and male rats at doses >= 2 mL/kg bw did not gain body weight. The following findings were observed at necropsy of dead animals: stomach abnormalities (discoloration, brown liquid or gas, gastric collapse), discoloured lungs, brown to dark red livers, intstines, and kidneys. Necropsy of the surving animals revealed the following effects: small stomachs and spleens; rough stomach surfaces, adhesion of the stomach to liver or spleen, enlarged and mottled red kidneys.Using the moving average method an LD50 value of 1.87 mL/kg bw (corresponding to 1750 mg/kg bw) was calculated for male and female rats.

Cross-reference 2

Reason / purpose:

reference to same study

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Principles of method if other than guideline:

Inhalation Risk Test

GLP compliance:

not specified

Test type:

fixed concentration procedure

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals and environmental conditions:

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

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: animal chmaber
- Exposure chamber volume: 100 to 152 L
- Method of holding animals in test chamber: in cage
- Source and rate of air: air in the exposure chamber
- System of generating atmosphere: 100 g of the test material was placed in the sealed exposure chamber for approx. 18 hours. A mixing fan periodically agitated the chamber atmosphere to aid in distribution of the vapour. Oxgen was added, as needed, to maintain approx. 20% of oxygen.

Analytical verification of test atmosphere concentrations:

not specified

Duration of exposure:

6 h

Concentrations:

saturation

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: on days 0 (prior to treatment), 7, and 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

none

Key result

Sex:

male/female

Dose descriptor:

LC0

Effect level:

8 375 mg/m³ air (nominal)

Based on:

test mat.

Exp. duration:

6 h

Remarks on result:

other: There were no signs of toxicity and no gross lesions at necropsy.

Mortality:

No death during exposure or thereafter

Clinical signs:

other: None noted

Body weight:

Increase in males from 280 g on day 0 to 339 g on day 14. No significant body weight gain in females (237 g before treatment, 244 g at day 14 (all values means).

Gross pathology:

No gross lesions noted

Interpretation of results:

GHS criteria not met

Conclusions:

Male and female rats (5 per sex) survived a 6-hour whole body exposure to a saturated vapour atmosphere. There were no deaths within the 14-day observation period. There were no clinical signs of toxicity at any time, or gross lesions at terminal sacrifice. According to EC/1272/2008 table 3.1.1 the substance is not acutely toxic after inhalation.

Executive summary:

Male and female rats (5 per sex) survived a 6-hour whole body exposure to a saturated vapour atmosphere. There were no deaths within the 14-day observation period. There were no clinical signs of toxicity at any time, or gross lesions at terminal sacrifice.

Cross-reference 3

Reason / purpose:

reference to same study

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Similar to guideline study

Reason / purpose:

reference to same study

Reason / purpose:

reference to same study

Reason / purpose:

reference to same study

Reason / purpose:

reference to same study

Qualifier:

equivalent or similar to

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL

Duration of treatment / exposure:

1) 4 hours and
2) 3 minutes

Observation period:

14 days

Number of animals:

6

Details on study design:

TEST SITE
- Area of exposure: no data
- % coverage: no data
- Type of wrap if used: gauze patch with impervious sheeting

REMOVAL OF TEST SUBSTANCE
- Washing (if done): excess material is removed after the 4-hr or 3-min contact period

SCORING SYSTEM: Table of Draize (as in OECD 404)

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 5 hr after skin contact

Score:

1.3

Max. score:

4

Reversibility:

not reversible

Remarks:

Skin necroses were seen at the 5-hr reading in 3 rabbits. Irreversible skin alterations were seen in all animals at 14 after 14 days post treatment.

Remarks on result:

other: 3-minute skin contact

Irritation parameter:

edema score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: 5 hr after skin contact

Score:

2.5

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: 3-minute skin contact

Irritant / corrosive response data:

At the 5-hr skin examination and thereafter, skin necrosis was seen in most rabbits exposed for 3 minutes and all animals exposed for 4 hours. Full thickness necrosis was confirmed at termination. No reading was made within 1 hour after skin contact.

Skin reactions after 4-hr contact period (means of 6 animals); 0.5 mL dosed

Time post skin contact	Erythema (mean score)	Edema (mean score)	Specific effects(selected effects)) N=necrosis U = ulceration
5 hours	2.0	4.0	N (all animals)
1 day	2.5	4.0	N (all animals)
2 days	3.0	4.0	N (all animals) U (1 animal)
3 days	2.8	4.0	N (all animals) U (5 animals)
7 days	1.0	-	N (all animals) U (5 animals)
10 days	0.3	-	N (all animals) U (5 animals)
14 days	0.0	-	N (all animals) U (5 animals)

 

Skin reactions after a 3-minute contact period (means of 6 animals); 0.5 mL dosed

 

Time post skin contact	Erythema (mean score)	Edema (mean score)	Specific effects(selected effects) N=necrosis U = ulceration D = desquamation
5 hours	1.3	2.5	N (3 animals)
1 day	2.3	3.8	N (3 animals)
2 days	2.2	3.0	N (4 animals)
3 days	0.0	2.3	N (4 animals)
7 days	0.0	0.0	N (5 animals) D (5 animals)
10 days	0.0	0.0	N (5 animals) D (5 animals)
14 days	0.0	0.0	D (5 animals)

 

Interpretation of results:

Category 1B (corrosive) based on GHS criteria

Conclusions:

2-methylbuytric acid caused full thickness skin necrosis in 5 animals after a 3-minute contact to the intact rabbit skin. Necrosis was already seen in 3 animals at the first reading at 5 hours after skin contact. The substance is at least a CAT1B skin corrosive; classification as a CAT 1A corrosive is also possible but cannot be judged because this would require a reading within one hour after the skin contact (Regulation (EC) No. 1272/2008).

Executive summary:

Skin irritating/corrosive properties of the test item were investigated in New Zealand rabbits in a study similar to OECD TG 404. Rabbits were exposed for 3 minutes or 4 hours toward 0.5 mL of the undiluted test item and observed for up to 14 days. Full thickness skin corrosion was observed in animal after 3 minutes or 4 hour exposure and was evident already at the first reading at 5 hours after the start of the exposure.

Cross-reference 4

Reason / purpose:

reference to same study

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

other information

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

Significant methological deficencies: instilled volume insufficient (0.005 mL)

Reason / purpose:

reference to same study

Reason / purpose:

reference to same study

Reason / purpose:

reference to same study

Reason / purpose:

reference to same study

Qualifier:

equivalent or similar to

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

insufficient test materila (0.005 mL)

GLP compliance:

not specified

Strain:

New Zealand White

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.005 mL

Duration of treatment / exposure:

single dose

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

6

Details on study design:

SCORING SYSTEM: Darize (OECD 405)

Irritant / corrosive response data:

Instillation of 0.005 mL per eye resulted in moderate to severe corneal opacity in 6/6 animals. Iris was not affected and minor to severe conjunctival irritation developed in 6/6 rabbits. A purulent discharge developed after 48 hours in all animals, followed by necrosis of the nictating membrane in all animals. By day 21, 4/5 eyes appeared normal.

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information

Conclusions:

Under the conditions of the test the substance caused severe eye damage including necrosis.

Data source

Reference

Reference Type:

other: TSCAT submission

Title:

Unnamed

Year:

1992

Report Date:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Test material

Reference

Name:

Unnamed

Type:

Constituent

Test material form:

liquid

Details on test material:

- Name of test material (as cited in study report): 2-methylbutyric acid
- Analytical purity: 97.9%
- Impurities (identity and concentrations): valeric acid 1.8%; unidentified 0.3%
- Stability under test conditions: shell life > 6 months
- Storage condition of test material: room temperature

Test animals

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Weight at study initiation: between 2.0 and 3.0 kg

Administration / exposure

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Type of wrap if used: occlusive

REMOVAL OF TEST SUBSTANCE
- Washing: excess fluid removed
- Time after start of exposure: 24 hr

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
males: 0.5. 1.0, 2.0, 2.83, and 4.0 mL/kg bw
females: 0.5. 1.0, 1.41, 2.0, and 4.0 mL/kg bw
- Constant volume or concentration used: no

Duration of exposure:

24 hr

Doses:

males: 0.5. 1.0, 2.0, 2.83, and 4.0 mL/kg bw (i.e. 468, 936, 1872, 2649, 3744 mg/kg bw)
females: 0.5. 1.0, 1.41, 2.0, and 4.0 mL/kg bw (i.e. 468, 936, 1320, 1872, 3744 mg/kg bw)

No. of animals per sex per dose:

5

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: on days 0 (prior to treatment), 7, and 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, skin reactions at one hour, 7 days and 14 days after contact period.

Statistics:

LD50 value was calculated using teh moving average method (Thompson, 1947; Weil, 1983)

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

2 228 mg/kg bw

Based on:

test mat.

Remarks on result:

other: The applied volume was converted to mg/kg bw using a density of 0.936 kg/L

Effect levels 2

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

1 367 mg/kg bw

Based on:

test mat.

Remarks on result:

other: The applied volume was converted to mg/kg bw using a density of 0.936 kg/L

Mortality:

Moast deaths occurred at 1.5 hours to one day; one female died at 3 days. A few affected survivors recovered after one to three days.

Males: all animals (5/5) of the two highest dose groups (4 and 2.83 mL/kg bw) died, no mortalities occurred at lower dose levels
Females: all animals (5/5) of the two highes dose groups (4 and 2 mL/kg bw) died, two animals (2/5) of the mid dose group (1.41 mL/kg bw) died, no animals died at the lower dose groups

Clinical signs:

Local efefcts: erythema, edema, necrosis, desquamation, alopecia, ulceratins, scabs.
Sluggishness, unsteady gait, prostration, red discharge on the perinasal and perianal fur were among the signs of toxicity noted.

Body weight:

Males: body weight gain in groups at 0.5 and 1.0 mL/kg bw; no body weight gain at 2.0 mL/kg.
Females: body weight gain in groups at 0.5 and 1.0 mL/kg bw; no body weightgain at 1.41 mL/kg bw. Complete mortality at higher dose levels.

Gross pathology:

Findings included a few discoloured lungs; red to tan livers; grey stomachs; stomachs and/or intestines with black foci; small instetsines fileld with red liquid (in 2); dark purple or red kidneys; dark red thymuses; and bladders filled with blood.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute dermal LD50 was 2228 mg/kg bw in male and 1367 mg/kg bw in female rabbits.Changes noted in internal organs suggest dermal absorption of the acid and irritation/corrosion of the organs.

Executive summary:

Dermal toxicity was investigated in a study similar to OECD TG 405. Male and female white rabbits (n=5 per sex per dose) were exposed for 24 h against different doses of test substance up to 4 mL/kg bw (3744 mg/kg bw by calculation on basis of substance density of 0.936 kg/L). Exposed animals showed local effects like erythema, edema, necrosis, desquamation, alopecia, ulceratins, and scabs. Clinical sign in treated animals were sluggishness, unsteady gait, prostration, red discharge on the perinasal and perianal fur. Body weight did not increase in animals which received more than 1 mL/kg bw. At necropsy discoloured lungs, red to tan livers, grey stomachs, stomachs and/or intestines with black foci, small instetsines fileld with red liquid (in 2), dark purple or red kidneys, dark red thymuses, and bladders filled with blood were observed. The acute dermal LD50 was 2228 mg/kg bw in male and 1367 mg/kg bw in female rabbits.