Phosphoric acid, mono C16-20 (branched, even numbered) alkyl esters

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 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

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Phosphoric acid, mono C16-20 (branched, even numbered) alkyl esters

Molecular formula:

unspecified

IUPAC Name:

Phosphoric acid, mono C16-20 (branched, even numbered) alkyl esters

Test material form:

liquid: viscous

Details on test material:

- Name (as cited in the report): SAT 150011
- Chemical name: Phosphoric acid, mono C16-20 (branched, even numbered) alkyl esters
- Appearance: clear, light amber liquid
- Purity: 70.2% mono and diesters
- Batch No.: RP GW 915 MU/11.06.2015
- Expiry date: 01/06/2018
- Date of receipt : 07/07/2016
- Storage conditions : ambient (15-25 °C)

Specific details on test material used for the study:

- Triskelion Dispense number : 16014D

Test animals / tissue source

Species:

other: Isolated chicken eyes

Strain:

other: not applicable

Details on test animals or tissues and environmental conditions:

Approximately 7 weeks old, male or female chickens (ROSS, spring chickens), body weight range approximately 1.5-2.5 kg, were used as eye donors.
Heads of these animals were obtained from poultry slaughterhouse v.d. Bor, Nijkerkerveen, the Netherlands.
Heads of the animals were cut off immediately after sedation of the animals by electric shock and incision of the neck for bleeding, and before they reached the next station on the process line.
The heads were placed in small plastic boxes on a bedding of paper tissues moistened with isotonic saline.
Next, they were transported to the testing facility. During transportation, the heads were kept at ambient temperature.

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):

240 minutes

Number of animals or in vitro replicates:

3

Details on study design:

- 3 eyes for the test item
- 3 eyes as positive control treated with BAC (Benzalkonium chloride) 5%
- 1 eye as negative control treated with physiological saline NaCl 0.9%

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

percent corneal swelling

Remarks:

mean

Run / experiment:

SAT 150011

Value:

19

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Remarks:

mean

Run / experiment:

SAT 150011

Value:

2.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Remarks:

Small pieces of the test substance adhered to one cornea and a large piece adhered to another cornea; the (white) remnants of the test substance could be removed at t= 75 min.

Results 3

Irritation parameter:

fluorescein retention score

Remarks:

mean

Run / experiment:

SAT 150011

Value:

2.3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 4

Irritation parameter:

in vitro irritation score

Remarks:

mean

Run / experiment:

SAT 150011

Value:

121

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

RESULTS

An overall summary of the results of the positive control and test substance based on maximum mean values for corneal swelling, corneal opacity, and fluorescein retention, the irritation categories assigned to the parameters, the irritation index and the regulatory classifications are presented in Table 1.

Table 1: Summary results of the slit-lamp examination (mean values)

Test material	Swelling [%]	Opacity	Fluorescein retention	Irritation Cat. (per eye)	Irritation Index	EU CLP classification
SAT 150011	19	2.8	2.3	III; IV; III	121	2
BAC 5% (pos. control)	41	3.0	3.0	IV; IV; IV	161	1

Upon the first observation of effects at t = 30 min, it appeared that small pieces of the test substance had adhered to cornea no. 1 and a large piece to cornea no. 5. At t = 75 min, the (white) remnants of the test substance could be removed. SAT 150011 caused corneal effects consisting of slight or severe corneal swelling (mean of 19%), moderate-to-severe or severe opacity (mean score of 2.8), and severe fluorescein retention (mean score of 3.0).

The negative control eye did not show any corneal effect and demonstrated that the general conditions during the tests were adequate.

The positive control BAC 5% caused severe corneal effects and demonstrated the ICE test valid to detect severe eye irritants.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

Applying the classification criteria of the ICE, the following irritation classifications can be assigned:
- Category 2A:“Irritant/causes eye irritation” (UN-GHS classification);
- Category 2:“Irritating to eyes” (EU-CLP classification).

Executive summary:

SAT 150011 was evaluated neat for eye irritation potential in the Isolated Chicken Eye (ICE) test. In addition, the test included a negative control (saline) and a positive control (BAC 5%). Chicken eyes were obtained from slaughter animals used for human consumption. The isolated chicken eyes were exposed to a single application of 30 μL for 10 seconds followed by a 20 mL saline rinse. Three main parameters were measured to disclose possible adverse eye effects: corneal thickness (expressed as corneal swelling), corneal opacity and fluorescein retention of damaged epithelial cells.

Upon the first observation of effects at t = 30 min, it appeared that small pieces of the test substance had adhered to cornea no. 1 and a large piece to cornea no. 5. At t = 75 min, the (white) remnants of the test substance could be removed. SAT 150011 caused corneal effects consisting of slight or severe corneal swelling (mean of 19%), moderate-to-severe or severe opacity (mean score of 2.8), and severe fluorescein retention (mean score of 3.0).

The negative control eye did not show any corneal effect and demonstrated that the general conditions during the tests were adequate. The positive control BAC 5% caused severe corneal effects and demonstrated the ICE test valid to detect severe eye irritants.

Applying the classification criteria of the ICE, the following irritation classifications can be assigned:

- Category 2A:“Irritant/causes eye irritation” (UN-GHS classification);

- Category 2:“Irritating to eyes” (EU-CLP classification).

SAT 150011 was a borderline case with Category 1.