Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10 May 2017 - 15 January 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study conducted in accordance with International guidelines and GLP. All guideline validity criteria were met.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Version / remarks:
26 July 2013
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Version / remarks:
Council Regulation (EC) No. 440/2008, 30 May 2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium octane-1-sulphonate monohydrate
EC Number:
226-195-4
EC Name:
Sodium octane-1-sulphonate monohydrate
Cas Number:
5324-84-5
Molecular formula:
C8H17O3S.Na
IUPAC Name:
sodium octane-1-sulphonate monohydrate
impurity 1
Reference substance name:
Sodium 1,8-disulphonate
IUPAC Name:
Sodium 1,8-disulphonate
impurity 2
Chemical structure
Reference substance name:
Sodium hexadecane-1-sulphonate
EC Number:
239-100-6
EC Name:
Sodium hexadecane-1-sulphonate
Cas Number:
15015-81-3
Molecular formula:
C16H34O3S.Na
IUPAC Name:
Sodium Hexadecane sulphonate
impurity 3
Chemical structure
Reference substance name:
Sodium sulphate
EC Number:
231-820-9
EC Name:
Sodium sulphate
Cas Number:
7757-82-6
Molecular formula:
O4S.2Na
IUPAC Name:
sodium sulphate
impurity 4
Chemical structure
Reference substance name:
Sodium chloride
EC Number:
231-598-3
EC Name:
Sodium chloride
Cas Number:
7647-14-5
Molecular formula:
ClNa
IUPAC Name:
sodium chloride
impurity 5
Chemical structure
Reference substance name:
Benzoic acid
EC Number:
200-618-2
EC Name:
Benzoic acid
Cas Number:
65-85-0
Molecular formula:
C7H6O2
IUPAC Name:
65-85-0
impurity 6
Reference substance name:
Unknown impurities
Molecular formula:
Not applicable
IUPAC Name:
Unknown impurities
Test material form:
solid
Details on test material:
Storage: ambient (5 °C – 30 °C), dark, dry
Specific details on test material used for the study:
RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: Assumed stable
- Solubility and stability of the test substance in the solvent/vehicle: Suspended and assumed stable.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not reported

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item was suspended with physiological saline (0.9 % NaCl) to give a 20 % concentration w/w.
- Preliminary purification step (if any): N/A
- Final dilution of a dissolved solid, stock liquid or gel: 20 % solution w/w
- Final preparation of a solid: N/A

FORM AS APPLIED IN THE TEST (if different from that of starting material) : Applied as a liquid

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable) : N/A

OTHER SPECIFICS: N/A

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: A. Moksel AG, Buchloe, Germany - abattoir
- Number of animals: Not reported
- Characteristics of donor animals (e.g. age, sex, weight): Cattle aged between 16 - 45 months
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): The eyes were removed after slaughter, completely immersed in Hanks’ Balanced Salt Solution (containing penicillin and streptomycin (1 % v/v)) in a container on ice and transported on the same day to the testing facility.
- Time interval prior to initiating testing: Same day as slaughter
- indication of any existing defects or lesions in ocular tissue samples: Only corneas free of defects were used.
- Indication of any antibiotics used: Penicillin and streptomycin used to transport eyes to testing facility.

Test system

Vehicle:
physiological saline
Remarks:
B. Braun Melsungen, Batch # 17031412, Expiry 26 May 2017
Controls:
yes, concurrent vehicle
yes, concurrent positive control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): 20 %

VEHICLE
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): 0.9 % NaCl
- Lot/batch no. (if required): 17031412
- Purity: Not reported
Duration of treatment / exposure:
4 hours ± 5 mins
Observation period (in vivo):
N/A
Duration of post- treatment incubation (in vitro):
After the exposure period, the corneas were rinsed with minimum essential medium with phenol red (MEM) and RPMI 1640 without phenol red (RPMI) (once the corneas were free of test item). Corneas were then incubated in RPMI and illuminance measurements recorded. Corneas were then incubated in fresh RPMI containing 1 mL of a 5 mg/mL sodium flurescein solution for 90 minutes.
Number of animals or in vitro replicates:
3 corneas per treatment group
Details on study design:
SELECTION AND PREPARATION OF CORNEAS: On arrival at the test facility the eyes were carefully examined for defects. Only corneas free from such defects were used.

QUALITY CHECK OF THE ISOLATED CORNEAS: The opacity of each cornea was measured using an opacitometer. Three corneas with illuminance readings approximately equivalent to the median illuminance of all corneas were selected as negative controls. Only corneas with an initial illuminance reading of I > I0 / 1.1651 lux were used for the assay.

NUMBER OF REPLICATES: 3 per treatment group.

NEGATIVE CONTROL USED: Vehicle control used below.

SOLVENT CONTROL USED (if applicable): The vehicle control substance was 0.9% sodium chloride solution.

POSITIVE CONTROL USED: The positive control substance was a 20 % solution of imidazole in 0.9 % NaCl.

APPLICATION DOSE AND EXPOSURE TIME: 4 hours ± 5 mins

TREATMENT METHOD:

After recording the initial illuminance measurements , 750 µL of the test item solution or control solution was incubated with each prepared cornea for 4 hours (± 5 mins) at 32 ± 1 ºC. The test item (or control solution) was rinsed from the cornea using MEM. Once free of test item, the cornea was rinsed and incubated with RPMI - a second illuminance reading was then taken along with a viaual inspection of the cornea condition. After the initial reading, the cornea was incubated with fresh RPMI containing 1 mL of a 5 mg/mL sodium fluoroscein solution for 90 mins at 31 ± 1 ºC. After 90 mins the solution was removed and the optical density measured using a spectrophotometer at 490 nm wavelength.

POST-INCUBATION PERIOD:
- See above

REMOVAL OF TEST SUBSTANCE
- See above

POST-EXPOSURE INCUBATION
- See above


METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Opaciometer.
- Corneal permeability: Passage of sodium fluorescein dye measured with the aid of spectrophotometry (OD 490).
- Others (e.g, pertinent visual observations, histopathology): (please specify) Pertinent visual inspection of cornea

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: please specify if the decision criteria as indicated in the TG was used. As per OECD 437.

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Value:
208.5
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: All test item cornea showed complete opacity of the tissue and whiteish discolouration of the corneas.

DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes, IVIS score for the negative and positive controls fell within 2 standard deviations of the laboratries current historical mean (n=23)

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Range of historical values if different from the ones specified in the test guideline: See above

Any other information on results incl. tables

Table 2:       Opacity

 

Cornea No.

Test Group

Initial Opacity

Final Opacity

Change of Opacity

Corrected Opacity

1

Negative Control

1.42

1.38

-0.04

N/A

2

1.64

1.78

0.15

3

1.42

2.77

1.35

Mean

1.49

1.98

0.49

SD

0.13

0.72

0.76

4

Positive Control

2.27

79.48

77.21

76.72

5

2.01

85.31

83.31

82.82

6

2.35

91.75

89.40

88.91

Mean

2.21

85.51

83.30

82.82

SD

0.18

6.13

6.09

6.09

7

Test Item

0.11

174.49

174.38

173.89

8

0.01

157.90

157.89

157.40

9

0.70

166.78

166.08

165.59

Mean

0.27

166.39

166.12

165.63

SD

0.37

8.31

8.25

8.25

 

Table 3:       Permeability

 

Cornea No.

Test Group

OD490

Corrected OD490Value

1

Negative Control

0.006

N/A

2

0.015

3

0.015

Mean

0.012

SD

0.005

4

Positive Control

0.963

0.951

5

0.944

0.932

6

1.106

1.094

Mean

1.004

0.992

SD

0.089

0.089

7

Test Item

2.355

2.343

8

2.860

2.848

9

3.395

3.383

Mean

2.870

2.858

SD

0.520

0.520

 

Table 4:       In Vitro Irritation Score (IVIS)

 

Cornea No.

Test Group

Corrected Opacity

Corrected OD490

IVIS

1

Negative Control

-0.04

0.006

0.67

2

0.15

0.015

3

1.35

0.015

Mean

0.49

0.012

SD

0.76

0.005

4

Positive Control

76.72

0.951

97.70

5

82.82

0.932

6

88.91

1.094

Mean

82.82

0.992

SD

6.09

0.089

7

Test Item

173.89

2.343

208.50

8

157.40

2.848

9

165.59

3.383

Mean

165.63

2.858

SD

8.25

0.520

 

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
Under the condition of this study, the test item, produced an IVIS score of 208.50 and therefore has the potential to induce serious eye damage. Sodium octane-1-sulphonate can be classified as UN GHS Category 1.
Executive summary:

OECD 437 (2017) - The Bovine Corneal Opacity and Permeability (BCOP) test was conducted using sodium octane 1-sulphonate in accordance with OECD Guideline 437 "Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage" (2013).

 

Test item was dissolved in physiological saline (0.9 % NaCl) to form a 20 % solution concentration. Triplicate corneas had their opacity measured and were incubated in the test solution for 4 hours ± 5 mins before being rinsed with minimal essential medium (with phenol red). After being further rinsed with RPMI (without phenol red) the opacity of each cornea was again measured. The permeability of the corneas was determined by measuring RPMI solution containing sodium fluorescein that has passed through the cornea membrane, after being suspended for 90 mins in a horizontal position, using a spectrophotometer. A negative and positive control group, each containing 3 corneas, were also prepared and treated using the same methodology.

 

For the test item, each of the treated corneas showed complete opacity of the tissue and whitish discolouration of the corneas. The negative and positive control values were in concordance with historical values and all guideline validity criteria were satisfied.

 

Under the conditions of this study, and with an IVIS score of 208.5, sodium octane-1-sulphonate showed the potential to induce serious eye damage. According to the guideline evaluation criteria, the test item is classified as UN GHS Category 1 for serious eye damage/ eye irritation.