2,2'-azobis[2-methylpropionamidine] dihydrochloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-Azo bis(2-amidinopropane)dihydrochloride

IUPAC Name:

2,2'-Azo bis(2-amidinopropane)dihydrochloride

Constituent 2

Reference substance name:

2,2'-azobis[2-methylpropionamidine] dihydrochloride

EC Number:

221-070-0

EC Name:

2,2'-azobis[2-methylpropionamidine] dihydrochloride

Cas Number:

2997-92-4

Molecular formula:

C8H18N6.2ClH

IUPAC Name:

2,2'-diazene-1,2-diylbis(2-methylpropanimidamide) dihydrochloride

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Vazo(R) 56 WSP
- Substance type: organic
- Physical state: solid
- Analytical purity: > 97 %
- Lot/batch No.: M0200
- Stability under test conditions: assumed to be stable for six months from date of receipt
- date for receipt: 10 November 1998
- Storage condition of test material: Stored at room temperature in the dark from date of receipt and at 4°C in the dark from 10 December 1998

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River UK Ltd, Margate, Kent, England
- Age at study initiation: least 12 weeks
- Weight at study initiation: 2.4 to 3.3 kg
- Housing: individual in metal cages
- Diet: Special Diet Services STANRAB (P) SQC pellets ad libitum
- Water: ad libitum
- Acclimation period: 11 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 19 °C
- Humidity (%): 31 to 44 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml

The weight of the test substance which when gently compacted occupied a volume of 0.1 ml was measured.
For this measurment, a syringe was used:
Syringe Weight of 0.1 ml test material (mg)
1 76
2 69
3 85
Mean 77

Duration of treatment / exposure:

none

Observation period (in vivo):

4 or 7 days, see table

Number of animals or in vitro replicates:

1+2

Details on study design:

SCORING SYSTEM: Draize scoring system
TOOL USED TO ASSESS SCORE: hand-slit lamp

One animal was treated in advance of the other two, again to ensure that if a severe response was produced, no further animals would be exposed (pilot animal)
Examination of the eyes was made after 1 hour and after 1, 2 and 3 days after instillation. Additional observations were made for all three animals 4 days after instillation and for two animals 5 through 7 days after instillation.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.66

Max. score:

4

Reversibility:

fully reversible within: 7 d

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 d

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 d

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Results 6

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 7 d

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.66

Max. score:

4

Reversibility:

fully reversible within: 7 d

Results 10

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Results 11

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2.66

Max. score:

3

Reversibility:

fully reversible within: 7 d

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 d

Irritant / corrosive response data:

Corneal opacification (Grade 1) was observed in two animals.
A diffuse, beefy red colouration of the conjunctivae was ssen in one animal. A diffuse, crimson colouration of the conjunctivae was seen in the remaining two animals. In addition, above normal swelling to partial eversion of eyelids was observed in two animals. Ocular reactions gradually ameliorated and had resolved in all instacnes by four or seven days after instillation.

Any other information on results incl. tables

Ocular reactions

 

Rabbit no. & sex	Region of eye			One hour	Day after instillation
					1	2	3	4	7
1954 Male*	Cornea		Density	0	0	1	1	0	0
			Area	0	0	1	1	0	0
	Iris			0	0	0	0	0	0
	Conjunctiva	Redness		2	2	2	2	1	0
		Chemosis		1	1	1	1	1	0
1955 Male	Cornea	Density		0	0	0	0	0	-
		Area		0	0	0	0	0	-
	Iris			0	0	0	0	0	-
	Conjunctiva	Redness		2	2	1	1	0	-
		Chemosis		0	0	0	0	0	-
1956 Male	Cornea	Density		0	1	1	0	0	0
		Area		0	1	1	0	0	0
	Iris			0	0	0	0	0	0
	Conjunctiva	Redness		2	3	3	2	2	0
		Chemosis		0	2	2	2	1	0

* pilot animal

- Not applicable

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The substance is classified as mildly irritating to the eye. In two of three animals, scores >= 2 for Conjunctiva Redness were found after 24, 48 and 72 hours which is fully reversible within 7 days.