Reaction products of 1-(substitutedphenyl)urea coupled with diazotated potassium sodium substituted-5-{[2-(substituted)ethyl]sulfonyl}benzenesulfonate, further condensed with 2,4,6-trichloro-1,3,5-triazine, further converted with disubstituted benzene-1,4-disulfonic acid in aq. sodium hydroxide

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was conducted between 14 April 2016 and 21 April 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 28 July 2015

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Test animals

Species:

rabbit

Strain:

New Zealand White

Remarks:

Two male rabbits

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS (UK) Limited, Leicestershire, UK.
- Age at study initiation: 12 to 20 weeks old
- Weight at study initiation: 2.53 or 2.73 kg
- Housing :individually housed in suspended cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period:at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23 °C
- Humidity (%): 30 to 70%
- Air changes (per hr): at least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs/12 hrs

IN-LIFE DATES: From: 14 April 2016 To: 21 April 2016

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

not required

Amount / concentration applied:

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

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

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

2

Details on study design:

TEST SITE
- Area of exposure: 2.5 cm x 2.5 cm
- Type of wrap if used: To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset and the animals were returned to their cages for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): gentle swabbing with cotton wool soaked in 74% industrial methylated spirits
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
1, 24, 48 and 72 hours

SCORING SYSTEM:

EVALUATION OF SKIN REACTIONS
Erythema and Eschar Formation
- No erythema : 0
- Very slight erythema (barely perceptible): 1
- Well-defined erythema: 2
- Moderate to severe erythema: 3
- Severe erythema (beef redness) to eschar formation preventing grading of erythema: 4

Edema Formation
- No edema: 0
- Very slight edema (barely perceptible): 1
- Slight edema (edges of area well-defined by definite raising): 2
- Moderate edema (raised approximately 1 millimeter): 3
- Severe edema (raised more than 1 millimeter and extending beyond the area of exposure): 4

Any other skin reactions and clinical signs of toxicity, if present, were also recorded.
An additional observation was made on Day 7 to assess the reversibility of skin reactions.
Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

DATA EVALUATION
The scores for erythema and edema at the 24 and 72 Hour readings were totaled for the two test rabbits (8 values) and this total was divided by 4 to give the primary irritation index of the test item. The test item was classified according to the following scheme devised by Draize, J.H. (1959):

- Method of calculation: Draize, J.H. (1959)
Primary Irritation Index Classification of Irritancy
0 Non-irritant
> 0 to 2 Mild irritant
> 2 to 5 Moderate irritant
> 5 to 8 Severe irritant

If irreversible alteration of the dermal tissue is noted in any rabbit, as judged by the Study Director, which include ulceration and clear necrosis or signs of scar tissue, the test item is classified as corrosive to rabbit skin. Classification according to Draize may, therefore, not be applicable.
The results were also interpreted according to the Globally Harmonized System of Classification and Labelling of Chemicals.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: Immediately

Score:

0

Remarks on result:

other: Orange colored staining of the test site

Results 2

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 1 hour

Score:

0

Remarks on result:

other: Orange colored staining of the test site

Results 3

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Orange colored staining of the test site

Results 4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

48 h

Score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Orange colored staining of the test site, Loss of skin elasticity

Results 5

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

72 h

Score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Orange colored staining of the test site, Loss of skin elasticity, Loss of skin flexibility

Results 6

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

7 d

Score:

0

Results 7

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: Immediately

Score:

0

Remarks on result:

other: Orange colored staining of the test site

Results 8

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 1 hour

Score:

0

Remarks on result:

other: Orange colored staining of the test site

Results 9

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24 h

Score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Orange colored staining of the test site

Results 10

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

48 h

Score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Orange colored staining of the test site, Loss of skin elasticity

Results 11

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

72 h

Score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Orange colored staining of the test site, Loss of skin elasticity, Loss of skin flexibility

Results 12

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

7 d

Score:

0

Results 13

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: Immediately

Score:

0

Results 14

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 1 hour

Score:

0

Results 15

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24 h

Score:

2

Reversibility:

fully reversible within: 7 days

Results 16

Irritation parameter:

edema score

Basis:

animal #1

Time point:

48 h

Score:

2

Reversibility:

fully reversible within: 7 days

Results 17

Irritation parameter:

edema score

Basis:

animal #1

Time point:

72 h

Score:

2

Reversibility:

fully reversible within: 7 days

Results 18

Irritation parameter:

edema score

Basis:

animal #1

Time point:

7 d

Score:

0

Results 19

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: Immediately

Score:

0

Results 20

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 1 hour

Score:

0

Results 21

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24 h

Score:

2

Reversibility:

fully reversible within: 7 days

Results 22

Irritation parameter:

edema score

Basis:

animal #2

Time point:

48 h

Score:

2

Reversibility:

fully reversible within: 7 days

Results 23

Irritation parameter:

edema score

Basis:

animal #2

Time point:

72 h

Score:

1

Reversibility:

fully reversible within: 7 days

Results 24

Irritation parameter:

edema score

Basis:

animal #2

Time point:

7 d

Score:

0

Results 25

Irritation parameter:

overall irritation score

Basis:

animal: 1 and 2

Time point:

other: 24 and 72 Hours

Score:

15

Reversibility:

fully reversible within: 7 days

Results 26

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

3.8

Reversibility:

fully reversible within: 7 days

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

The test item produced a primary irritation index of 3.8 and was classified as a moderate irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.
Well-defined erythema and slight edema were noted at both treated skin sites at the 24 and 48 Hour observations with well-defined erythema and very slight to slight edema noted at the 72 Hour observation.
Both treated skin sites appeared normal at the 7-Day observation.

Other effects:

- Other adverse local effects: Loss of skin flexibility and/or elasticity was noted at both treated skin sites at the 48 and 72 Hour observations. Both treated skin sites appeared normal at the 7-Day observation.
- Other adverse systemic effects: One animal showed body weight loss and the other animal showed expected gain in body weight during the study.

Any other information on results incl. tables

Orange colored staining of the test site, not preventing evaluation of skin responses, was noted at both treated skin sites during the study. The test substance is a dark orange powder.

Individual Skin Reactions

Skin Reaction	Observation Time (following patch removal)	Individual Scores		Total
		Rabbit Number and Sex
		75409Male	75412Male
Erythema/Eschar Formation	Immediately	0STA	0STA	(0 )
	1 Hour	0STA	0STA	( 0 )
	24 Hours	2STA	2STA	4
	48 Hours	2STALe	2STALe	( 4 )
	72 Hours	2STALeLf	2STALeLf	4
	7 Days	0	0	( 0 )
Edema Formation	Immediately	0	0	( 0 )
	1 Hour	0	0	( 0 )
	24 Hours	2	2	4
	48 Hours	2	2	( 4 )
	72 Hours	2	1	3
	7 Days	0	0	( 0 )
Sum of 24 and 72‑Hour Readings (S)            :              15
Primary Irritation Index (S/4)                          :              15/4 = 3.8
Classification                                                      :              MODERATEIRRITANT

(   ) =    Total values not used for calculation of primary irritation index

STA =   Orange colored staining of the test site

Le =      Loss of skin of elasticity

Lf =       Loss of skin flexibility

Individual Body Weights and Body Weight Change

Rabbit Number and Sex	Individual Body Weight (kg)		Body Weight Change (kg)
	Day 0	Day 7
75409Male	2.73	2.84	0.11
75412Male	2.53	2.50	-0.03

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item produced a primary irritation index of 3.8 and was classified as a moderate irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted. however, the test item was classified as non irritant according to the Globally Harmonized System of Classification and Labelling of Chemicals, since the effects found were fully reversible by the end of 7 days and mean scores for erythema and edema were <2.3.

Executive summary:

Introduction

The study was performed to assess the irritancy potential of the test item to the skin of the New Zealand White rabbit.

Results

A single 4‑Hour, semi‑occluded application of the test item to the intact skin of two rabbits produced well-defined erythema, slight edema and loss of skin elasticity and flexibility. Both treated skin sites appeared normal at the 7‑Day observation.

Conclusion

The test item produced a primary irritation index of¨3.8 and was classified as amoderateirritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted. However, the test item was classified as non irritant according to the Globally Harmonized System of Classification and Labelling of Chemicals, since the effects found were fully reversible by the end of 7 days and mean scores for erythema and edema were <2.3.