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Description of key information

Skin irritation of Chloroacetamide was tested in vivo in a key irritation study in rabbits, resulting in mean scores of  1.1 and 0.5 for erythema and edema, and some additional changes that were reversible within 7 days.  Eye irritation was tested in vivo in a key irritation study in rabbits, resulting in mean scores  1.7, 1.0, 2.9 and 3.0 for cornea, iris, conjunctival redness and chemosis, respectively, and additional changes that were all revesible within 21 days. Based on these data, Category 2 classification for eye irritation was warranted.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1993
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted under GLP application, and it was done according to valid methods. Therefore it is considered relevant, reliable and adequate for classification.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
other: Albino New Zealand White Chbb: NZW (SPF)
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Chemisch pharmazeutische Fabrik Dr. Karl Thomae GmbH, Biberach (Germany), SPF-breeding
- Age at study initiation: Ca. 3 - 5 months
- Weight at study initiation: 2,5 - 2,8 kg
- Housing: Individual cages (battery cages)
- Diet: Altromin 2123 Haltungsdiät – Kaninchen by Altromin-GmbH, Lage/Lippe (Germany), ad libitum; also hay, ca. 15 g daily
- Water: Deionised chlorinated water, automatic watering, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18± 3 °C
- Humidity (%):55 ± 20%
- Air changes (per hr): Fully air-conditioned rooms
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 1993-07-06 To: 1993-07-13
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
physiological saline
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 500 mg chloroacetamide
- Concentration (if solution): moistened with 0.25 mL 0.9% NaCl solution

Duration of treatment / exposure:
4 hour(s)
Observation period:
7 days
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: dorsal area of the trunk
- Type of wrap if used: The mixed substance was applied on a wound plaster with 2.5 X 2.5 cm cellulose pad and the plaster strips were attached to the prepared skin site under a semiocclusive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): carefully with lukewarm tap water
- Time after start of exposure: 4 hours

SCORING SYSTEM:
See "Any other information on material and methods incl. tables"
Irritation parameter:
other: erythema and eschar formation
Basis:
mean
Time point:
other: mean; ,24, 48, 72 h after removal of plaster
Score:
1.1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
other: erythema and eschar formation
Basis:
animal #1
Time point:
other: mean; 24, 48, 72 h after removal of plaster
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7days
Irritation parameter:
other: erythema and eschar formation
Basis:
animal #2
Time point:
other: mean; 24, 48, 72 h after removal of plaster
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
other: erythema and eschar formation
Basis:
animal #3
Time point:
other: mean; 24, 48, 72 h after removal of plaster
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
mean
Time point:
other: mean; 24, 48, 72 h after removal of plaster
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: mean; 24, 48, 72 h after removal of plaster
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: mean; 24, 48, 72 h after removal of plaster
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: mean; 24, 48, 72 h after removal of plaster
Score:
1.3
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
The animals showed 1hour until 3 days after application very slight erythema to moderate to severe erythema, as well as very slight to moderate edema. Additional in one animal the skin was dry, rough and with coarse scales.

Table 1. Results

Sex: Female

Animal Number

Erythema (time after decontamination)

Mean score*

30-60 min

1 day

2 days

3 days

7 days

96

2

2

1

0

0

1.0

97

2

1

0

0

0

0.3

98

3

2

2

2

0

2.0

 OVERALL MEAN ERYTHEMA (24, 48, 72h): 1.1

Animal Number

Edema (time after decontamination)

Mean score*

30-60 min

1 day

2 days

3 days

7 days

96

2

1

0

0

0

0.3

97

1

0

0

0

0

0.0

98

3

2

1

1

0

1.3

OVERALL MEAN EDEMA (24, 48. 72h): 0.5

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The Skin irritation test in rabbits has shown that Chloroacetamide according to the present study is assessed as slightly irritating.
Executive summary:

A dorsal area of the trunk of approximately 25 cm2 of 3 rabbits was shaved ca. 24 hours before the start of the test . Each animal (intact skin) was applied with 500 mg Chloroacetamide (made into a paste with 0.25 mL of 0.9% NaCl solution). The mixed substance was applied on a wound plaster with 2.5 X 2.5 cm cellulose pad and the plaster strips were attached to the prepared skin site under a semiocclusive bandage. After 4 hours exposure, the remaining test substance was washed carefully with lukewarm tap water. Observations were done at 30-60 minutes and at 24, 48 and 72 hours after removal of the plaster. As there were some other findings in 1 animal (skin suface: coarse scales, dry, rough), additional observation was done up to 7 days. All changes were reversible within 7 days. Althoug Chloroacetamide was assessed as slightly irritating, there is no reason for classifcation.

The Skin irritation test in rabbits has shown that Chloroacetamide according to the present study is assessed as slightly irritating.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1993
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted under GLP application, and it was done according to valid methods. Therefore it is considered relevant, reliable and adequate for classification.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
other: Albino, Chbb: New Zealand White (SPF)
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Chemisch pharmazeutische Fabrik Dr. Karl Thomae GmbH, Biberach (Germany), SPF-Breeding
- Age at study initiation: Ca. 3 - 5 months
- Weight at study initiation: 2.9-3.4 kg
- Housing: Individual (battery cages)
- Diet: Altromin 2123 Haltungsdiät – Kaninchen by Altromin-GmbH, Lage/Lippe (Germany), ad libitum, also hay, ca. 15 g daily
- Water: Deionised chlorinated water, automatic watering, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3 °C
- Humidity (%):55 ± 20 %;
- Air changes (per hr): fully air-conditioned rooms
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 1993-07-27 To: 1993-08-19

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg per eye (left eye only, right eye used as control)
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Eyes with discharges or assessed with fluorescein-sodium solution were rinsed with physiological saline before each assessment.

SCORING SYSTEM: See under 'Any other information on material and methods incl. tables '

TOOL USED TO ASSESS SCORE: fluorescein
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
1.7
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
no. 93
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
1.7
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Remarks:
no. 96
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
1
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Remarks:
no. 98
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
2.3
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
1
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
no. 93
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
1
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Remarks:
no. 96
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
1
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Remarks:
no. 98
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
1
Max. score:
2
Irritation parameter:
conjunctivae score
Remarks:
, redness
Basis:
mean
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
2.9
Max. score:
3
Irritation parameter:
conjunctivae score
Remarks:
, redness
Basis:
animal #1
Remarks:
no. 93
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
2.7
Max. score:
3
Irritation parameter:
conjunctivae score
Remarks:
, redness
Basis:
animal #2
Remarks:
no. 96
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
3
Max. score:
3
Irritation parameter:
conjunctivae score
Remarks:
, redness
Basis:
animal #3
Remarks:
no. 98
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
3
Max. score:
3
Irritation parameter:
conjunctivae score
Remarks:
, chemosis
Basis:
mean
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
3
Max. score:
4
Irritation parameter:
conjunctivae score
Remarks:
, chemosis
Basis:
animal #1
Remarks:
no. 93
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
3
Max. score:
4
Irritation parameter:
conjunctivae score
Remarks:
, chemosis
Basis:
animal #2
Remarks:
no. 96
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
2.7
Max. score:
4
Irritation parameter:
conjunctivae score
Remarks:
, chemosis
Basis:
animal #3
Remarks:
no. 98
Time point:
other: mean, 24, 48, 72 h p.appl.
Score:
3.3
Max. score:
4
Irritant / corrosive response data:
1 Hour until 7 days after application the conjunctivae of the animals showed a significant hyperemia of some blood vessels up to a diffuse deep red color and a slight swelling to a swelling with lids more than half-closed. The iris was partially reddened. The cornea showed partially diffuse up to pearlescent areas of opacity . 14 Days after application in one animal there was still a marked hyperemia of some conjunctival blood vessels. The iris of this animals was reddened and the cornea still showed light visible translucent opacity areas. The irritation symptoms were accompanied with clear, colorless or white slimy discharge. Furthermore partially bleeding from the conjunctiva and nictitating membrane were observed. 21 Days after application all signs of irritation were reversible.

Based on the individual scores at 24, 48 and 72 hours the following mean values were calculated:

Corneal opacity Iris score
All animals: 1.7 All animals: 1.0
Animal 93: 1.7 Animal 93: 1.0
Animal 96: 1.0 Animal 96: 1.0
Animal 98: 2.3 Animal 98: 1.0
Conjunctiva redness Conjunctiva chemosis
All animals: 2.9 All animals: 3.0
Animal 93: 2.7 Animal 93: 3.0
Animal 96: 3.0 Animal 96: 2.7
Animal 98: 3.0 Animal 98: 3.3

Table 1. Individual readings

Animal Number

Elapsed Time

Cornea

Iris

Conjunctiva

Opacity

Area

Redness

Chemosis

Discharge

93

1 hour

1

4

1

2

2

2

 

1 day

2

2

1

3

4

3

 

2 days

2

2

1

3

3

2

 

3 days

1

1

1

2

2

1

 

7 days

0

0

0

1

1

0

 

14 days

0

0

0

0

0

0

96

0.9 hours

1

1

0

2

2

1

 

1 day

1

2

1

3

3

2

 

2 days

1

1

1

3

3

1

 

3 days

1

1

1

3

2

1

 

7 days

0

0

0

1

1

0

 

14 days

0

0

0

0

0

0

 

21 days

0

0

0

0

0

0

98

0.9 hours

1

1

0

2

2

2

 

1 day

2

3

1

3

4

3

 

2 days

2

3

1

3

3

2

 

3 days

3

3

1

3

3

2

 

7 days

3

3

1

2

2

0

 

14 days

2

1

1

1

0

0

 

21 days

0

0

0

0

0

0

 

Interpretation of results:
other: Category 2
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Classification: irritating
Executive summary:

In 3 rabbits, 100 mg chloroacetamide was applied in the conjunctival sac of the left eye. The untreated eye served as control. The assessments of the eyes were made at 1, 24, 48 and 72 hours after application of test substance. At the evaluation time points 24 and 72 hours after application the cornea was additionally examined for damage after instillation of fluorescein-sodium solution. Mean scores for cornea, iris, conjunctival redness and chemosis were 1.7, 1.0, 2.9 and 3.0, respectively. As there were still effects on the eyes at 72 hours, an additional observation was done at 7, 14 and 21 days.

After 21 days all symptoms of irritation were reversible. Based on this study Chloroacetamide was evaluated as irritant.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Additional information

In a key primary skin irritation study (Hoechst, 1993a) in 3 New Zealand White rabbits, 500 mg Chloroacetamide (made into a paste with 0.25 mL 0.9% saline solution) was applied to the intact skin for 4 hours under semiocclusive bandage. Mean scores were 1.1 for erythema/eschar formation and 0.5 for edema, with individual scores varying between 0.3 -2 and 0-1.3, respectively. As there were some other findings in 1 animal (skin surface: coarse scales, dry, rough), additional observation was done up to 7 days. All changes were reversible within 7 days. Although the substance was reported as slightly irritating, there is no reason for classification. In a supporting study (Hoechst, 1958) no signs of irritation of Chloracetamide were observed at the depilated flank skin of rabbits at concentrations up to 10%. In another supporting study with a low concentration (5%) of a different formulation (70% active ingredient; 30% sodium benzoate), primary irritation index was 0.2 (IBR, 1981a).

In a key eye irritation study of 3 New Zealand White rabbits (Hoechst, 1993b), 100 mg Chloroacetamide was applied in the conjunctival sac of the left eye. The untreated eye served as control. The assessments of the eyes were made at 1, 24, 48 and 72 hours after application and rinsing of test substance. After 1 to 7 hours the animals displayed severe reddening and swelling of the conjunctivae. In addition, corneal clouding was seen. As there were still effects at 72 hours, additional observations were done at 7, 14 and 21 days. Mean scores for cornea, iris, conjunctival redness and chemosis were 1.7, 1.0, 2.9 and 3.0, respectively. Additional changes observed were all reversible within 21 days. Based on this, Chloroacetamide was evaluated as irritant for the eye. In a supporting study (Hoechst, 1958) no signs of irritation were observed at the eyes of rabbits at concentrations up to 10%. In another supporting study in rabbits with a low concentration (5%) of a different formulation (70% active ingredient; 30% sodium benzoate), primary irritation index was 0.7 (IBR, 1981b).



Justification for selection of skin irritation / corrosion endpoint:
Key study; the study was conducted under GLP application, and it was done according to valid methods. Therefore it is considered relevant, reliable and adequate for classification.

Justification for selection of eye irritation endpoint:
Key study; the study was conducted under GLP application, and it was done according to valid methods. Therefore it is considered relevant, reliable and adequate for classification.

Effects on skin irritation/corrosion: slightly irritating

Effects on eye irritation: irritating

Justification for classification or non-classification

Based on these results and according to the EC Directive (No.93/21/EEC) and CLP (No. 1272/2008 of 16 December 2008), Chloroacetamide does not have to be classified and has no obligatory labelling requirement for skin irritation.

Chloroacetamide was classified according the EU labelling regulations Commision Directive 93/21/EEC with symbol 'Xi R36' and risk phrase R 36 'IRRITATING TO EYES'. According to CLP regulation (No. 1272/2008 of 16 December 2008), Category 2 classification is proposed with signal word 'WARNING' and hazard statement H319 'CAUSES SERIOUS EYE IRRITATION'.