Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts

Eye irritation

Administrative data

Endpoint:

eye irritation, other

Remarks:

in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1999-09-23 to 1999-10-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP 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)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

the European Economic Communities (EEC) Guideline in the Official Journal of the European Communities [92/69, Annex V, B5 (1992)l

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: the Japanese Agricultural Chemicals Laws and Regulations Testing Guidelines for Toxicology Studies published by the Society of Agricultural Chemicals Industry, under the auspices of the (MAFF) [59 NohSan No. 4200 (1985)

Deviations:

no

GLP compliance:

yes

Remarks:

in compliance with the United States Environmental Protection Agency (EPA) Good Laboratory Practice Regulations (40 CFR Parts 160 and 792)

Test material

Test material information

Constituent 1

Reference substance name:

Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts

EC Number:

946-212-5

Molecular formula:

Too complex

IUPAC Name:

Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts

Test material form:

liquid: viscous

Details on test material:

- Substance type: benzenesulphonate
- Physical state: dark brown, viscous liquid
- Analytical purity: Test article purity data are the responsibility of sponsor
- Impurities (identity and concentrations): not provided
- Composition of test material, percentage of components: UVCB
- Lot/batch No.: None provided
- Stability under test conditions: Test article stability data are the responsibility of the sponsor
- Storage condition of test material: Ambient
- Date received: April 28,1999

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Covance Research Products, Inc. Denver, PA
- Age at study initiation: Young adult
- Weight at study initiation: 3295 to 3615 grams at initiation of dosing
- Housing: Individually in suspended wire mesh cages
- Diet: PMI Nutrition International, Inc. Certified Rabbit LabDiet@ 5322 was offered at approximately 150 g per day during the study.
- Water (e.g. ad libitum): Municipal water was provided ad libitum.
- Acclimation period: for a minimum of seven days prior to initiation of dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): (68.2 - 69.1°F; 20 - 21 °C)
- Humidity (%): 30.2-64.1
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
Prior to use, the original container of the test article was agitated to insure a homogeneous mixture. A sufficient amount of test article was dispensed into a labeled storage container and a stir bar was added. The test article was stirred continuously throughout use.
The test article was placed directly into the cupped lower conjunctival sac of each rabbit's right (test) eye. The eyelids were held closed for approximately one second after instillation. Left eyes were manipulated in an identical manner to simulate the dosing of the right eyes.
- Amount(s) applied (volume or weight with unit): 0.1 mL

Duration of treatment / exposure:

single, unwashed exposure.

Observation period (in vivo):

at approximately one, 24, 48 and 72 hours after dosing and on days 4, 7, 14 and 21, if irritation persisted.

Number of animals or in vitro replicates:

6 (Two males and four females)

Details on study design:

REMOVAL OF TEST SUBSTANCE: Washing: no
SCORING SYSTEM: presented in "Any other information on materials and methods incl. tables".
TOOL USED TO ASSESS SCORE: fluorescein, ophtalmoscope

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Time point:

other: 24, 48 and 72 hour

Score:

12.2

Max. score:

110

Reversibility:

fully reversible within: 21 days

Remarks on result:

other: at one-hour post-instillation

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

other: no findings

Results 3

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

other: no findings

Results 4

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

other: mean of 24, 48 and 72-h readings

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

Results 5

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

other: mean of 24, 48 and 72-h readings

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 21 days

Results 6

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

other: mean of 24, 48 and 72-h readings

Score:

1.7

Max. score:

2

Reversibility:

fully reversible within: 4 days

Results 7

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #4

Time point:

other: mean of 24, 48 and 72-h readings

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

Results 8

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #5

Time point:

other: mean of 24, 48 and 72-h readings

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Results 9

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #6

Time point:

other: mean of 24, 48 and 72-h readings

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: mean of 24, 48 and 72-h readings

Score:

0.7

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: mean of 24, 48 and 72-h readings

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: mean of 24, 48 and 72-h readings

Score:

0.7

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Results 13

Irritation parameter:

chemosis score

Basis:

animal #4

Time point:

other: mean of 24, 48 and 72-h readings

Score:

0.7

Max. score:

2

Reversibility:

fully reversible within: 48 hours

Results 14

Irritation parameter:

chemosis score

Basis:

animal #5

Time point:

other: mean of 24, 48 and 72-h readings

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Results 15

Irritation parameter:

chemosis score

Basis:

animal #6

Time point:

other: mean of 24, 48 and 72-h readings

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Irritant / corrosive response data:

No animals vocalized upon instillation of the test article. All animals were noted with conjunctival irritation. Iridal irritation was noted for one animal. There were no corneal findings. All irritation completely subsided by day 21. The left (control) eyes were free of evidence of ocular irritation and other findings for the duration of the study.

Other effects:

There were no deaths or remarkable body weight changes during the study period.

Any other information on results incl. tables

Table 1. Individual ocular irritation scores

			Examination intervals
Animal	Sex	Tissue	1h	24h	48h	72h*	4 days	7 days*	14 days*	21 days*	Mean (24-48-72 h)
29139	M	Cornea (O-A)	0-0	0-0	0-0	0-0	0-0	0-0	0-0	-	0.0
		Iris	1	0	0	0	0	0	0	-	0.0
		Conjunctiva (R-C-D)	2-2- 2	2-1-2	2-1-0	2-0-0	2-0-0	2-0-0	0-0-0	-	2.0 (R) - 0.7 (C)
29143	M	Cornea (O-A)	0-0	0-0	0-0	0-0	0-0	0-0	0-0	0-0	0.0
		Iris	0	0	0	0	0	0	0	0	0.0
		Conjunctiva (R-C-D)	2-2-2	2-2-0	2-1-0	2-0-0	2-0-0	2-0-0	1-0-0	0-0-0	2.0 (R) – 1.0 (C)
29158	M	Cornea (O-A)	0-0	0-0	0-0	0-0	0-0	-	-	-	0.0
		Iris	0	0	0	0	0	-	-	-	0.0
		Conjunctiva (R-C-D)	2-1-1	2-1-0	2-1-0	1-0-0	0-0-0	-	-	-	1.7 (R) - 0.7 (C)
29148	F	Cornea (O-A)	0-0	0-0	0-0	0-0	0-0	0-0	0-0	-	0.0
		Iris	0	0	0	0	0	0	0	-	0.0
		Conjunctiva (R-C-D)	2-2-2	2-2-0	2-0-0	2-0-0	2-0-0	1-0-0	0-0-0	-	2.0 (R) - 0.7 (C)
29155	F	Cornea (O-A)	0-0	0-0	0-0	0-0	0-0	0-0	-	-	0.0
		Iris	0	0	0	0	0	0	-	-	0.0
		Conjunctiva (R-C-D)	2-2-2	2-2-0	2-1-0	2-0-0	2-0-0	0-0-0	-	-	2.0 (R) – 1.0 (C)
29156	F	Cornea (O-A)	0-0	0-0	0-0	0-0	0-0	0-0	0-0	-	0.0
		Iris	0	0	0	0	0	0	0	-	0.0
		Conjunctiva (R-C-D)	2-2-2	2-2-0	2-1-0	2-0-0	2-0-0	1-0-0	0-0-0	-	2.0 (R) – 1.0 (C)
Total			73	44	34	22	20	12	2.0	0.0
Mean			12.2	7.3	5.7	3.7	3.3	2.0	0.3	0.0
Mean group scores		Cornea									0.0
		Iris									0.0
		Conjunctiva									2.0 (R) – 0.9 (C)

*Sodium fluorescein in solution applied;

O = opacity;
A = area;

R = redness;

C = chemosis;

D = discharge;

h = hours;

M = male;

F = female

- = no observation made anymore 

Applicant's summary and conclusion

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to the classification criteria outlined in the section 3.3.2.3.2.2. (Guidance on the Application of CLP criteria, 2013), if a substance cause positive but reversible reactions such as corneal opacity (score ≥ 1), iritis (score ≥ 1), conjunctival redness (score ≥ 2) or conjunctival oedema (chemosis) (score ≥ 2) in at least 4 of 6 tested animals from gradings at 24, 48 and 72 hours after application, the classification as Eye Irritant (Category 2) is assigned. The average irritation scores over 24, 48, and 72 hours for both corneal opacity and iritis were 0.0 in each of six animals. The mean conjunctival chemosis scores (swelling) for each animal were all under the cut-off value of 2.0 for this time period. However, the positive responses (score of 2.0) were observed in 5 of 6 animals regarding conjunctival erythema (redness). The irritation reactions were completely reversible within 21 days. Based on this observation, the target substance (Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts) is classified as Eye Irritant (Category 2, H319) according to Regulation (EC) No. 1272/2008.

Executive summary:

The primary ocular irritation potential of C20 -24 sodium sulfonate (TBN=448) was evaluated in this study with New Zealand White rabbits. There was one group of six albino rabbits that received a single, unwashed exposure. Each 0.1 mL dose of the test article was instilled into the lower conjunctival sac of the right eye. The eyelid was held closed for approximately one second and released. The left eye was manipulated in a similar manner as the right eye and served as a contralateral control. The eyes were examined for ocular reactions in accordance with the method of Draize at approximately one, 24, 48 and 72 hours after dosing and on days 4, 7, 14 and 21, if irritation persisted. Sodium fluorescein was used to aid in revealing possible corneal damage at 72 hours and on days 7, 14 and 21. There were no deaths or remarkable body weight changes during the study period. All animals were noted with positive (grade 2) conjunctival irritation. Iridal irritation was noted for one animal. There were no corneal findings. All irritation completely subsided by day 21. The Maximum Average Score for C20 -24 sodium sulfonate (TBN=448) was 12.2 at one-hour post-instillation.

The following table presents the mean scores calculated from individual scores:

Conjunctiva -      Redness -      Oedema	24 and 72 hours	24,48 and 72 hours
	1.9	2.0
	0.8	0.9
Iris	0.0	0.0
Cornea	0.0	0.0