3',5'-dichloro-4'-ethyl-2'-hydroxypalmitanilide

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 1989, july 14th to 1989, july 29th

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study performed according to the Test Guidelines described in the EEC Directive 84/449/EEC

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

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:

3',5'-dichloro-4'-ethyl-2'-hydroxypalmitanilide

EC Number:

406-200-8

EC Name:

3',5'-dichloro-4'-ethyl-2'-hydroxypalmitanilide

Cas Number:

117827-06-2

Molecular formula:

C24H39Cl2NO2

IUPAC Name:

N-(3,5-dichloro-4-ethyl-2-hydroxyphenyl)hexadecanamide

Details on test material:

Identification UC—136
Description White powder
Batch Number Not indicated by the sponsor
Purity 98,5%
Bulk Density 0.7 g/ml
Instructions for test article storage At room ternperature in the dark in a tightly sealed container
Stability of test article Stable for one year maximal under storage conditions
expiry date: .January 5, 1990
Safety precautions Gloves, goggles and face mask were considered sufficient to ensure personnel health and safety.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Test system
Species Rabbit, New Zealand White (SPF—Quality)
Rationale Recognised by international guidelines as the recommended test system (e.g. EPA, OECD, EEC).
Original Source Charles River Wiga, Sulzfeld, West—Germany.
Total number of animals 3 females
Age at start of treatment Approx. 10 weeks
Body weight at start of treatment 2846 — 3037 gram
Identification Ear tag.
Acclimatisation
At least five days under test conditions
Husbandry
Room No.: 9
Conditions
Standard Laboratory Conditions. Air—conditioned with 7.5—15 air changes per hour and hourly monitored environment with temperature 21 ± 3°C, relative humidity 30—85 %, 12 hours artificial fluorescent light / 12 hours dark.
Accomodation
Individually in cages with perf’orated floors and equipped with an automatic drinking system (ITL, Bergen, The Netherlands).
Diet
Pelleted standard LKK—20 rabbit maintenance diet (Hope Farms, Woerden, The Netherlands) approx. 100 g per day. Certificates of analysis are retained in the RCC NOTOX archives.
Water
Free access to tap—water, diluted with decalcified water. Certificates of analysis of tap—water are retained in the RCC NOTOX archives.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

70+/- 1 mg (equivalent to 0.1 ml) / animal

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 females

Details on study design:

The following numerical scoring system was used to score the eye lesions:
CORNEA
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Nacreous areas, no details of iris visible, size of pupil barely discernible 3
Opaque cornea, iris not discernible through the opacity 4
Area of cornea affected
No ulceration or opacity 0
One quarter or less but not zero 1
Greater than one quarter, but less than half 2
Greater than half, but less than three quarters 3
Greater than three quarters, up to whole area 4
IRIS
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any of these or any combination thereof, iris still reacting to light (sluggish reaction is positive) 1
No reacticn to light, haemorrhage, gross destruction (any or all of these) 2
CONJUNCTIVAE
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris):
Blood vessels normal 0
Some blood vessels definitely hyperaemic (injected) 1
Diffuse, crimson colour, individual vessels not easily discernible 2
Diffuse beefy red 3
Chemosis: lids and/or nictating membrane
No swelling 0
Any swelling above norrnal (includes nictating membrane) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids more than half closed 4
Di scharge:
No discharge 0
Any amount different from normal (does not include small amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs, just adjacent to lids 2
Discharge with moistening of the lids and hairs, in a considerable area around the eye 3
In case standard lighting was considered inadequate for observing minor effects, eye examinations were perf’orrned using a pocket flash light.
INTERPRETATION OF RESULTS:
The irritation scores and all other observations were recorded on data sheets and transcribed for compilation and analysis.
A Draize score was calculated for each day of observation, using the following formula (Draize et al., 3. Pharmacol. Exp. Ther. 82, 1944):
(5x corneal opacity grade x area of opacity grade) + (5x iridial injury grade)+ 2x (conjunctival redness grade + chemosis grade + discharge grade).
With the maximum score of the study (i.e. Draize score) a Kay and Calandra interpretation was obtained, using the table below and taking into account the time needed for healing (3. Society of Cosrnetic Chemists, Vol. B no. 6, 1962):
Draize score Tentative eye irritation rating to Kay and Cal andra.
0-0.5 Non—irritating N
0.5-2.5 Practically non—irritating PN
2.5-15 Minimally irritating M1
15-25 Mildly irritating M2
25-50 Moderately irritating M3
50-80 Severely irritating S
80-100 Extremely irritating E
100-110 Maximally irritating Mx
For borderline scores choose the higher rating.
The test resuits were evaluated according to the EEC general classification and labelling requirements for dangerous substances (Annex VI of the EEC
Council Directive 67/548/EEC as amended by Directive 83/467/EEC, September,1983).

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 48h

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 48 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 48h

Results 3

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 72 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 48h

Results 4

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 48h

Results 5

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 48 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 48h

Results 6

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 72 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 48h

Results 7

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24 hours

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48h

Results 8

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 48 hours

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48h

Results 9

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 72 hours

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48h

Results 10

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 48h

Results 11

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 48 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 48h

Results 12

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 72 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 48h

Irritant / corrosive response data:

The test article appeared to be minilnally irritating (Ml) to the rabbit eye (Kay and Calandra interpretation of the Draize score: 4).
Instillation of UC—136 into one of the eyes of each of three albino rabbits aff’ected the conjunctivae.
The irritation of the conjunctivae was reversible within 48 hours.
Treatment of the eyes with fluorescein 2%, 24 hours after test article instillation revealed no corneal epithelial damage in any of the animals.

Other effects:

COLOURATION
No staining of the eyes by the test article was observed.
CORROSION
No corrosion was observed.
TOXIC SYMPTOMS / MORTALITY
No toxic symptoms were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

slightly irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study, UC—136 resulted in adverse effects on the conjunctivae. The irritation of the conjunctivae was reversible within 48 hours.
No corrosion nor signs of systernic intoxication were observed.
The test article should be considered minimally irritating (Ml) to the rabbit eye (Kay and Calandra interpretation of’ the Draize score : 4).