Reaction products of “Fatty Acids, C16-18 and C18-unsatd., Me esters; sulfuric acid and sodium hydroxide

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of “Fatty Acids, C16-18 and C18-unsatd., Me esters; sulfuric acid and sodium hydroxide

EC Number:

951-929-1

Molecular formula:

Not available. UVCB

IUPAC Name:

Reaction products of “Fatty Acids, C16-18 and C18-unsatd., Me esters; sulfuric acid and sodium hydroxide

Test material form:

liquid

Specific details on test material used for the study:

Batch: 2018060002
pH: 7.60
moisture content: 30.29%

Test animals / tissue source

Species:

chicken

Details on test animals or tissues and environmental conditions:

The eyes collected from chickens obtained from a slaughterhouse (Etablissement Brun, 33820Etauliers, France) where they are killed for human consumption have been used for this assay.
The age and weight of the chickens used in this test method are that of spring chickens traditionally processed by a poultry slaughterhouse (i.e., approximately 7 weeks old, 1.5 - 2.5 kg).

Heads have been removed immediately after sedation of the chickens by electric shock, and incision of the neck for bleeding. The heads have been collected on 13 January 2020 at 8:28 am.

Because eyes were dissected in the laboratory, the intact heads were transported from the slaughterhouse at ambient temperature in plastic boxes humidified with towels moistened with
physiological saline.

The eyes were enucleated at Phycher on 13 January 2020 at 10:00 am

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

30 μL

Duration of treatment / exposure:

10 seconds

Duration of post- treatment incubation (in vitro):

Treated corneas were evaluated pretreatment and starting at 30, 75, 120, 180, and 240 minutes (± 5 minutes) after the post-treatment rinse.

Number of animals or in vitro replicates:

test item : 3 eyes
postive control : 3 eyes
negative control : 1 eye

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Run / experiment:

mean

Value:

0.3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

fluorescein retention score

Run / experiment:

mean

Value:

0.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

percent corneal swelling

Run / experiment:

mean

Value:

4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

A histopathological examination was performed. The study meets the acceptance criteria as the negative control item and the positive control item fall within the Not Classified and Severe irritant classification, respectively. Following the application of the test item RICOIL EN, there was very slight (1/2) erosion of the
superficial part of the superficial corneal epithelium in 1 out 3 eyes examined. According to these findings, the test item was classified as negative for GHS Category 1 for non-extreme pH (2 < pH < 11.5) detergents and surfactants.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In accordance with the Regulation (EC) No. 1272/2008, the results obtained under these experimental conditions enable to conclude that the test item does not require classification for eye irritation and serious eye damage as defined by the UN GHS (No Category).
No signal word and hazard statement are required.
However, these results are in contradiction with the results obtained with skin irritation in vitro (OECD 439). Indeed, the product has been classified in category 1 "Corrosive" or category 2 "Irritant" for its effects on the skin.
A histopathological examination was performed. The study meets the acceptance criteria as the negative control item and the positive control item fall within the Not Classified and Severe irritant classification, respectively. Following the application of the test item RICOIL EN, there was very slight (1/2) erosion of the superficial part of the superficial corneal epithelium in 1 out 3 eyes examined. According to these findings, the test item was classified as negative for GHS Category 1 for non-extreme pH (2 < pH < 11.5) detergents and surfactants.
Therefore, in accordance with the Regulation (EC) No. 1272/2008, the test item RICOIL EN is classified in the category “no prediction can be made”. It is not predicted as causing serious eye damage (Category 1) or as not classified for eye irritation/serious eye damage (No category) with the Isolated Chicken Eye test method.
As a conservative approach and in the absence of additional testing (in vitro and/or in vivo), the substance is classified as irritant to eye (category 2).

Executive summary:

The aim of the study was to evaluate the possible ocular corrosive or severe irritating effects of the test item after administration on enucleated chicken eyes.

The test item was applied, as supplied, at the dose of 30 μL, to 3 enucleated chicken eyes, during 10 seconds. Then the eyes were rinsed twice with 10 mL of physiological saline. Three eyes were treated in the same manner with a positive control and one eye with a negative control.

Damages by the test item were assessed by determination of corneal swelling, opacity, and fluorescein retention at 30, 75, 120, 180 and 240 minutes post-dose.

The experimental protocol was established in accordance with the O.E.C.D. Test Guideline No. 438 adopted 25 June 2018.

The ocular reactions observed in eyes treated with the test item were:

- maximal mean score of corneal opacity: 0.3, corresponding to ICE class I;

- mean score of fluorescein retention: 0.8, corresponding to ICE class II;

- maximal mean corneal swelling: 4%, corresponding to ICE class I.

The combination of the three endpoints for the test item was 2 x I, 1 x II.

The combination of the three endpoints for the positive control, 5% Benzalkonium chloride, was 3 x IV. Therefore, the positive control is classified as “Corrosive/Severe Irritant”, as expected.

The combination of the three endpoints for the negative control, physiological saline, was 3 x I. Therefore, the negative control is classified as “No Category”, as expected.

In accordance with the Regulation (EC) No. 1272/2008, the results obtained under these experimental conditions enable to conclude that the test item does not require classification for eye irritation and serious eye damage as defined by the UN GHS (No Category).

No signal word and hazard statement are required.

However, these results are in contradiction with the results obtained with skin irritation in vitro (OECD 439).

Indeed, the product has been classified in category 1 "Corrosive" or category 2 "Irritant" for its effects on the skin.

A histopathological examination was performed. The study meets the acceptance criteria as the negative control item and the positive control item fall within the Not Classified and Severe irritant classification, respectively. Following the application of the test item RICOIL EN, there was very slight (1/2) erosion of the superficial part of the superficial corneal epithelium in 1 out 3 eyes examined. According to these findings, the test item was classified as negative for GHS Category 1 for non-extreme pH (2 < pH < 11.5) detergents and surfactants.

Therefore, in accordance with the Regulation (EC) No. 1272/2008, the test item RICOIL EN is classified in the category “no prediction can be made”. It is not predicted as causing serious eye damage (Category 1) or as not classified for eye irritation/serious eye damage (No category) with the Isolated Chicken Eye test method.

As a conservative approach and in the absence of additional testing (in vitro and/or in vivo), the substance is classified as irritant to eye (category 2).