Esterification product of castor oil and tetrahydromethyl-1,3-isobenzofuranedione

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:

other: Test procedure according to validation study and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

The aim of this study was to evaluate the ocular irritancy of Multiester P97-463 as measured by its ability to induce adverse changes in the chorioallantoic membrane (CAM) of chicken eggs.
The study procedures were based on the following documents and follow international recommendations
- The Ocular Toxicity Working Group (OTWG) of the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) and the National Interagency Centre for the Evaluation of Alternative Toxicological Methods (NICEATM), Background Review Document (BRD): current status of in vitro test methods for identifying ocular corrosives and severe irritants: The Hen’s Egg Test - Chorioallantoic Membrane (HET-CAM) test method, March 2006.
- INVITTOX protocol 47. The HET-CAM test - Method of Spielmann and Liebsch, January 1992.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Esterification product of castor oil and tetrahydromethyl-1,3-isobenzofuranedione

EC Number:

700-064-6

Cas Number:

2105830-60-0

Molecular formula:

Not applicable as this is a UVCB substance

IUPAC Name:

Esterification product of castor oil and tetrahydromethyl-1,3-isobenzofuranedione

Constituent 2

Reference substance name:

[TN]Multiester P97-463[/TN][SPEC][/SPEC][AM]100 %[/AM]

IUPAC Name:

[TN]Multiester P97-463[/TN][SPEC][/SPEC][AM]100 %[/AM]

Details on test material:

Multiester P97-463
Batch No.: NB2890-61

Test animals / tissue source

Species:

other: HET-CAM

Strain:

not specified

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

300 mg test item
0.3 ml positive control (1% sodium dodecylphosphate prepared in physiological saline) and negative control (physiological saline)

Duration of treatment / exposure:

20 seconds

Observation period (in vivo):

5 minutes

Number of animals or in vitro replicates:

0

Details on study design:

The number of eggs used in the study was 3 per treatment group. On day 9 of incubation the air cells of the eggs were marked and cut off with a rotating saw blade. The uncovered inner membrane was moistened with physiological saline and subsequently the egg was kept at room temperature. Subsequently the physiological saline was removed and the inner egg membrane carefully eliminated using forceps to expose the CAM.
Three eggs were treated for 20 seconds with 300 mg Multiester P97-463 or with 0.3 ml of the negative or positive control solutions. After the treatment period the eggs were carefully rinsed with approximately 5 ml of physiological saline. The appearance of haemorrhage, vessel lysis and coagulation on the CAM was monitored and recorded over the following 280-second period.

Results from the three test method endpoints were evaluated separately for each egg. The irritancy score (IS) was calculated using the observed haemorrhage time, lysis time and coagulation time.

Results and discussion

In vivo

Results

Irritation parameter:

other: HET-CAM irritation score

Basis:

mean

Time point:

other: 301 s

Score:

0

Max. score:

21

Reversibility:

not specified

Remarks on result:

other: individual scores: see below

Other effects:

no other effects

Any other information on results incl. tables

The results from the three test method endpoints and irritancy scores of the individual eggs.  Egg Heamorrhage time (sec)  Lysis time (sec) Coagulation time (sec) irritancy score (IS)  Mean irritancy score (IS)                 Negative control  1 301  301  301  0.0     2 301  301  301  0.0     3 301  301  301  0.0               0.0                 1 % Sodium dodecyl sulphate  5 301  30  20  14.8     6 301  30  20  14.8     7 301  35  20  14.6               14.7                 Multiester P97 -463  15 301  301  301  0.0     16 301  301  301  0.0     17 301  301  301  0.0               0.0 Note: 301 seconds was noted when no effect was oberved after 5 minutes of observation.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: ICCVAM and NICEATM

Conclusions:

Classification: not irritating

Executive summary:

The mean in vitro irritancy score obtained with the negative control was less than 1 indicating that the negative control did not induce irritancy on the chorioallantoic membrane. The mean in vitro irritancy score of the positive control (1% (w/v) Sodium dodecyl sulphate) was within the historical control data range. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

The mean in vitro irritancy score obtained during a 280-second observation period after 20 seconds treatment with Multiester P97-463 was 0.

Finally, it is concluded that this test is valid and that Multiester P97-463 is not irritant in the Hen’s Egg Test – Chorioallantoic Membrane Test under the experimental conditions described in this report.

This report describes the ocular irritation properties of Multiester P97-463 on the chorioallantoic membrane of chicken eggs. The possible ocular irritancy of Multiester P97-463 was tested through topical application for 20 seconds.

The study procedures described in this report were based on recent HET-CAM documents.

Batch NB2890-61 of Multiester P97-463 was a very viscous amber liquid. The test substance was applied undiluted (300 mg) directly on top of the chorioallantoic membrane.

The mean in vitro irritancy score obtained with the negative control was less than 1 indicating that the negative control did not induce irritancy on the chorioallantoic membrane. The mean in vitro irritancy score of the positive control (1% (w/v) Sodium dodecyl sulphate) was 14.7 and within the historical control data range. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

The mean in vitro irritancy score obtained during a 280-second observation period after 20 seconds treatment with Multiester P97-463 was 0.

Finally, it is concluded that this test is valid and that Multiester P97-463 is not irritant in the Hen’s Egg Test – Chorioallantoic Membrane Test under the experimental conditions described in this report.