Reaction products of 1,6-diisocyanatohexane, oligomers with 2-[(2-methacryloyl)oxy]ethyl 6-hydoxyhexanoate and 2-hydroxyethyl methacrylate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 Aug 2009 to 26 Aug 2009

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well-documented study report which meets basic scientific principles performed in compliance with OECD GLP (1997)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Draft OECD Guideline for Testing of Chemicals: The Bovine Corneal Opacity and Permeability (BCOP) Test Method for Identifying Ocular Corrosives and Severe Irritants (December, 2008), INVITTOX (UK) protocol no. 98 "The Bovine Corneal Opacity and Permeabili

Deviations:

no

GLP compliance:

yes

Test material

Reference

Name:

Unnamed

Type:

Constituent

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report: MTDID 8191
- Physical state: Yellowish Liquid
- Analytical purity: >99%
- Lot/batch no.: MK12/048
- Expiration date of the lot/batch: 27 Jan 2010
- Storage condition of test material: 2 - 8 °C
- Other: Stable for 2 hours in DMSO at room temperature or 1 day in DMSO at 2 - 8 °C

Test animals / tissue source

Species:

other: Bovine

Strain:

not specified

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Odenwaldschlachthof Brensbach, D-64395, Brensbach
- Age at study initiation: Freshly isolated bovine cornea were stored at 2 - 8 °C and used on the same day as collection
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 32 °C
IN-LIFE DATES: From: August 11, 2009 To: August 11, 2009

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: Yes, concurrent (negative and positive control)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 1.0 mL
NEGATIVE CONTROL MATERIAL
- Identity: Sodium Chloride
- Amount(s) applied (volume or weight with unit): 1.0 mL
- Concentration (if solution): 0.9% (w/v)
POSITIVE CONTROL MATERIAL
- Identity: 2-Ethoxyethanol
- Amount(s) applied (volume or weight with unit): 1.0 mL
- Concentration (if solution): 100%
- Lot/batch no. (if required): 0001430432

Duration of treatment / exposure:

10 minutes +/- 30 seconds

Observation period (in vivo):

120 minutes

Number of animals or in vitro replicates:

3 corneas per treatment

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, Cell culture media (cMEM), minimum of 3 rinses
- Time after start of exposure: 10 minutes
SCORING SYSTEM:
- Opacity: The opacity value was calculated by subtracting the initial basal opacity for each cornea from the post-treatment opacity reading (t130 - t0).
- Permeability: The corrected optical density 490 nm (OD490) value for each positive control- and test article-treated cornea was calculated by subtracting the average value for the negative control-treated corneas from the original permeability value for each cornea.
- In vitro score for the negative control = opacity value +(15 x OD490 value)
- In vitro score for positive control and test article = (test item opacity value - negative control opacity value) + (15 x Corrected OD490 value)
- The mean in vitro score of the test article-treated corneas was calculated and compared to the ranges in Table 1 to interpret the results.
TOOL USED TO ASSESS SCORE: Opacity: opacitometer and Permeability: 0.5% fluorescein and spectrophotometer at OD490

Results and discussion

In vivo

Results

Irritation parameter:

other: In vitro score

Basis:

other: Calculated mean in vitro score based on opacity and permeability measurements.

Time point:

other: see remark

Remarks on result:

other: SEE REMARK in SECTION : REMARKS on results including tables.

Irritant / corrosive response data:

See remark in section : "Remarks on results including tables and figures"

Any other information on results incl. tables

REMARK for Section : Overall irritation/corrosion results:

Mean in vitro irritation scores were 1.81 and 77.29 for the negative control and positive control, respectively. These results indicated that the test was valid. The mean irritation score for MTDID 8191 was 1.40 which is considered non-irritating under the protocol-defined classification criteria. Individual cornea results are included in Table 2.

Table 2. Results After 10 Minutes Incubation Time
Test Group	Opacity value = Difference (t130-t0) of Opacity		Permeability at 490 nm (OD490)		In vitro Score	Mean in vitro Score	In vitro Scale Result
	Score	Mean	Score	Mean
Negative Control	1	1	0.049	0.054	1.74	1.81	Non irritant
Negative Control	1		0.061		1.92
Negative Control	1		0.052		1.78
Positive Control	52.00^*		0.472^*		59.08	77.29	Severe irritant
Positive Control	74.00^*		1.788^*		100.82
Positive Control	60.00^*		0.798^*		71.97
MTDID 8191	0.00^*		-0.004^*		-0.06	1.4	Non irritant
MTDID 8191	2.00^*		0.004^*		2.06
MTDID 8191	2.00^*		0.013^*		2.20
* Negative control mean values were subtracted from the positive control and test article values.

 

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: INVITTOX (UK) protocol no. 98 “The Bovine Corneal Opacity and Permeability Assay”, dated February 1994.

Conclusions:

Based on the results of this study, MTDID 8191 is not considered to be an eye irritant.

Executive summary:

OBJECTIVE: The corneal irritation and damage potential of MTDID 8191 (yellow liquid, batch MK 12/048) was tested in the Bovine Corneal Opacity and Permeability assay (BCOP).

METHODS: This study was conducted in compliance with OECD GLP (1997) and the Chemicals Act of Germany (2002). The study method was based on the OECD draft proposal for the BCOP assay (2008), INVITTOX (UK) protocol no. 98 (1994), and the SOP of Microbiological Associates Ltd. (1997). Bovine corneas (3 per group) were prepared in cell culture media. The corneas were mounted in an O-ring holder and incubated for at least 1 hour at 32 C prior to exposure. Corneas (3 per group) were treated with 0.75 mL of MTDID 8191, 2-ethoxyethanol (positive control), or 0.9% saline (negative control) and incubated for a 10 minute exposure period at 32 C. At the end of the exposure period, the corneas were rinsed with media and then incubated for approximately 120 minutes. Opacity was evaluated prior to treatment and 120 minutes after the initiation of exposure. Permeability was evaluated at 120 minutes after exposure using a fluorescein method. The mean in vitro irritation scores were calculated and the scores were classified according to protocol defined categories.

RESULTS: Mean in vitro irritation scores were 1.81 and 77.29 for the negative control and positive control, respectively. These results indicated that the test was valid. The mean irritation score for MTDID 8191 was 1.40 which is considered non-irritating under the protocol-defined classification criteria.

CONCLUSION: Based on the results of this study, MTDID 8191 is not considered an ocular irritant.