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Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was performed between 31 March 2000 and 17 May 2000.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report date:
2000

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Fatty acids, C18 unsaturated, reaction products with diethylenetriamine, acetate salts
EC Number:
938-649-5
Cas Number:
1469982-92-0
Molecular formula:
Not applicable (a generic molecular formula cannot be provided for this specific UVCB substance)
IUPAC Name:
Fatty acids, C18 unsaturated, reaction products with diethylenetriamine, acetate salts
Details on test material:
Sponsor's identification: PR 4659 SALT
Date received: 3 March 2000
Description: extremely viscous amber liquid
Storage conditions: room temperature in the dark.

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Rabbits supplied by David Percival, Moston, Sandbach, Cheshire, Uk.
- Age at study initiation: Twelve to sixteen weeks old.
- Weight at study initiation: 2.85 to 3.04 kg.
- Housing: The animals were individually housed in suspended metal cages.
- Diet (e.g. ad libitum): Free access to food was allowed throughout the study.
- Water (e.g. ad libitum): Free access to mains drinking water was allowed throughout the study.
- Acclimation period: Minimum period of five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): The temperature was set to achieve limits of 17 to 23°C.
- Humidity (%): The relative humidity was set to achieve limits of 30 to 70%.
- Air changes (per hr): Approximately fifteen changes per hour.
- Photoperiod (hrs dark / hrs light): The lighting was controlled by a time switch to give twelve hours continuous light and twelve hours darkness,

Test system

Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 ml oif test material at each test site.

Duration of treatment / exposure:
Single application for a maximum duration of 4 hours.
Observation period:
Approximately one hour following the removal of the patches, and 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation.
Number of animals:
3 animals.
Details on study design:
PROCEDURE:
One rabbit was initially treated. Three suitable test sites were selected on the back of the rabbit. At each test site a quantity of 0.5 ml of the test material was introduced under a 2.5 cm x 2.5 cm cotton gauze patch and placed in position on the shorn skin. Each patch was secured in position with a strip of surgical adhesive tape (BLENDERM: approximate size 2.5 cm x 4.0 cm). To prevent the animal interfering with the patches, the trunk of the rabbit was wrapped in an elasticated corset (TUBIGRIP) and the animal was returned to its cage for the duration of the exposure period.

One patch was removed at each of three time points: 3 minutes, 1 hour and 4 hours after application. Any residual test material was removed by gentle swabbing with cotton wool soaked in 74% Industrial Methylated Spirits.

After consideration of the skin reactions produced in the first animal, an additional animal was treated with 0.5 ml of test material. Two patches were applied to the back of the rabbit, and were allowed to remain in contact with the skin for a period of three minutes or four hours. After consideration of the skin reactions produced in the second animal, one further animal was similarly treated with 0.5 ml of the test material.

SCORING:
Approximately one hour following the removal of the patches, and 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the Draize scoring system (see below).

Any other skin reactions, if present, were also recorded.

Additional observations were made on Days 7 and 14 to assess the reversibility of skin reactions.









Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
erythema score
Basis:
mean
Remarks:
(animal 1)
Time point:
other: 24, 48 and 72 hours
Score:
3.7
Max. score:
4
Remarks on result:
other: 4-hour exposure period
Irritation parameter:
erythema score
Basis:
mean
Remarks:
(animal 2)
Time point:
other: 24, 48 and 72 hours
Score:
2.3
Max. score:
4
Remarks on result:
other: 4-hour exposure period
Irritation parameter:
erythema score
Basis:
mean
Remarks:
(animal 3)
Time point:
other: 24, 48 and 72 hours
Score:
4
Max. score:
4
Remarks on result:
other: 4-hour exposure period
Irritation parameter:
edema score
Basis:
mean
Remarks:
(animal 1)
Time point:
other: 24, 48 and 72 hours
Score:
4
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: 4-hour exposure period
Irritation parameter:
edema score
Basis:
mean
Remarks:
(animal 2)
Time point:
other: 24, 48 and 72 hours
Score:
2.7
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: 4-hour exposure period
Irritation parameter:
edema score
Basis:
mean
Remarks:
(animal 3)
Time point:
other: 24, 48 and 72 hours
Score:
2.3
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: 4-hour exposure period
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
other: Total of 24 and 72-hour readings for 3 animals (12 values) divided by 6
Time point:
other: 24 and 72-hour readings
Score:
6.3
Irritant / corrosive response data:
4-Hour Exposure Period:
The individual scores for erythema/eschar and oedema are given in Table 1 (see attached background material).
Mean values are given in Table 2 (see attached background material).

Well-defined erythema and moderate to severe oedema were noted at all treated skin sites at the 1-hour observation with moderate to severe erythema and moderate to severe oedema at the 24-hour observation and well-defined to severe erythema and slight to severe oedema at the 48 and 72-hour observations. Dermal haemorrhage was noted at one treated skin site at the 1-hour observation and at two treated skin sites at the 24, 48 and 72-hour observations. Blanching of the skin was noted at one treated skin site at the 24 and 48-hour observations. Light brown discolouration of the epidermis was noted at two treated skin sites at the 24 and 48-hour observations and at all treated skin sites at the 72-hour observation. Loss of skin elasticity and flexibility were noted at two treated skin sites at the 24, 48 and 72-hour observations.
Pale green discolouration, possibly superficial dermal necrosis, was scattered over one treated skin site at the 24, 48 and 72-hour observations. A hardened dark brown/black coloured scab or a hardened light brown coloured scab, which prevented an accurate evaluation of erythema and oedema, was noted at all treated skin sites at the 7-day observation. Glossy skin and/or reduced regrowth of fur were noted at all treated skin sites at the 14-day observation.

1-Hour Exposure Period:
The individual scores for erythema/eschar and oedema are given in Appendix I (see attached background material).

Very slight erythema and very slight oedema were noted at the treated skin site at the 24 and 48-hour observation with very slight erythema at the 72-hour observation.

The treated skin site appeared normal at the 7-day observation.

3-minute Exposure Period:
The individual scores for erythema/eschar and oedema are given in Appendix II (see attached background material).

Very slight erythema and very slight oedema were noted at two treated skin sites at the 1-hour observation. Very slight erythema was noted at all treated skin sites at the 24-hour observation, at two treated skin sites at the 48-hour observation and at one treated skin site at the 72-hour observation. Moderate desquamation was noted at two treated skin sites at the 7-day observation.

One treated skin site appeared normal at the 72-hour observation and two treated skin
sites appeared normal at the 14-day observation.


Applicant's summary and conclusion

Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test material, PR 4659 SALT, produced a primary irritation index of 6.3 and was classified as a severe irritant to rabbit skin according to the Draize classification scheme.
The test material produced positive criteria in 3/3 rabbits according to the EU labelling regulations and was classified as irritant to rabbit skin.


Executive summary:

Introduction:

A study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The method used followed the recommendations of the OECD Guidelines for Testing of Chemicals No. 404 "Acute Dermal Irritation/Corrosion" (adopted 17 July 1992) and Method B4 of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

Method and results:

A single 4-hour, semi-occluded application of the test material to the intact skin of three rabbits produced moderate to severe erythema and moderate to severe oedema.

Other skin reactions noted were light brown discolouration of the epidermis, blanching of the skin, haemorrhage of the dermal capillaries, loss of skin elasticity and flexibility and scabbing. Glossy skin and/or reduced regrowth of fur were noted at all treated skin sites at the 14-day observation.

 3 -minute and 1 -hour semi-occluded applications of the test material to the intact skin of one rabbit produced no corrosive effects.

Conclusion:

The test material, PR 4659 SALT, produced a primary irritation index of 6.3 and was classified as a severe irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

The test material produced positive criteria in 3/3 rabbits according to the EU labelling regulations and was classified as irritant to rabbit skin.