Fatty acids, C16-18 (even numbered), aluminum salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

4 Jul - 9 Aug 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Version / remarks:

adopted 7. September 2009

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Landesinstitut für Arbeitsschutz und Produktsicherheit, München

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C16-18 (even numbered), aluminum salts

EC Number:

939-582-4

Cas Number:

1471315-26-0

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Fatty acids, C16-18 (even numbered), aluminum salts

Details on test material:

- Name of test material (as cited in study report): Fatty acids, tallow, hydrogenated, aluminium salts
- Physical state: white solid powder
- Analytical purity: 84.33%
- Impurities (identity and concentrations): ash content 9.17%; humidity 0.75%; free fatty acids 14.13%; water-soluble salts 0.61%; aluminium 4.13%
- Lot/batch No.: C200752-001
- Expiration date of the lot/batch: 14.02.2014
- Storage condition of test material: at room temperature
- Other: CAS 91770-05-7

Test animals / tissue source

Species:

other: cattle

Strain:

other: bovine corneas

Details on test animals or tissues and environmental conditions:

TEST SYSTEM PREPARATION
- Source: isolated corneas obtained as a by-product from an abattoir from freshly slaughtered animals (Attenberger Fleisch GmbH & Co. KG, München)
- cornea preparation: on test day, fresh eyes were transported in HBSS containing Pen/Strep (5%) on ice. For cornea preparation, tissue surrounding the eyeball was carefully pulled away and corneas were excised leaving a 2 to 3 mm rim of sclera. Isolated corneas were stored in petri dishes containing HBSS.
- Cornea holders: corneas were mounted with the endothelial side against the O-ring of the posterior chamber, anterior chamber is positioned on top of of the cornea and tightend with screws, chamber of the cornea holder is filled with RPMI (without Phenol Red) containing 1% FBS and 2 mM L-glutamine, the posterior chamber was filled first, corneas were preincubated for one hour.

Test system

Vehicle:

other: sesame oil

Controls:

yes, concurrent vehicle

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): 20% in vehicle

Duration of treatment / exposure:

4 hours ± 5 min

Observation period (in vivo):

not applicable

Number of animals or in vitro replicates:

3 corneas each per treatment

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): after removal of the treatment medium, with MEM, once the medium was macroscopically free of test substance the corneas were rinsed with RPMI (without phenol red) and anterior chambers were filled with fresh RPMI for opacity measurements
- Time after start of exposure: 4 hours

EVALUATION OF RESULTS:
- mean opacity value: calculated by subtracting the initial opacity reading (before treatment) from the final reading (after exposure), these values were corrected by subtracting from each the average change in opacity from corresponding negative control, the mean then was calculated by averageing the corrected opacity values for each treatment group
- permeability measurements (fluorescein): mean OD490 for each treatment were calculated by substracting the blank from each value and by subtracting the average corrected negative control value from the corrected value of each treatment measurement, the mean then was calculated by averaging the corrected OD490 values by averaging the corrected values for each treatment group
- IVIS score: (In Vitro Irritation Score) - calculated by summing up the mean OD490 score multiplied by 15 with the mean opacity value for each treatment

TOOL USED TO ASSESS SCORE: fluorescein + measurement of optical density at 490 nm and visual inspection, after opacity measurement

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

other: IVIS Score

Basis:

other: negative control

Time point:

other: 4 hours

Score:

1.1

Results 2

Irritation parameter:

other: IVIS Score

Basis:

other: positive control

Time point:

other: 4 hours

Score:

216.51

Results 3

Irritation parameter:

other: IVIS Score

Basis:

other: negative control (vehicle)

Time point:

other: 4 hours

Score:

0.05

Results 4

Irritation parameter:

other: IVIS Score

Basis:

other: test item

Time point:

other: 4 hours

Score:

1.32

Irritant / corrosive response data:

no effects on the corneas were observed

Other effects:

no other effects observed

Any other information on results incl. tables

Table2: In Vitro Irritation Score

Cornea No.	Test Item	Corrected Opacity Value	Corrected OD490 Value	IVIS
1	Negative Control	1.00	0.087
2		0.00	0.089
3		-2.00	0.110
MV		-0.33	0.095	1.10
4	Positive Control	203.33	2.007
5		178.33	1.710
6		182.33	1.986
MV		188.00	1.901	216.51
10	Negative Control Sesame Oil	-1.00	0.063
11		0.00	0.030
12		-1.00	0.051
MV		-0.67	0.048	0.05
7	Test Item	0.67	0.026
8		1.67	-0.045
9		1.67	0.017
MV		1.33	-0.001	1.32

Applicant's summary and conclusion

Interpretation of results:

other: non-corrosive

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

According to Regulation (EC) No. 1272/2008, a substance can be considered a severe eye irritant (Serious eye damage Category 1/R41) based on a positive result in the Bovine Corneal Opacity and Permeability (BCOP) test. Negative in vitro corrosivity responses are not conclusive with respect to non-classification or classification as irritant (Category 2/R36) and shall therefore be subject to further evaluation."