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Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
other: read across from analogue substance
Adequacy of study:
key study
Study period:
16 April 2015 - 06 December 2015
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability 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

Constituent 1
Chemical structure
Reference substance name:
Amides, from diethylenetriamine and hydrogenated palm oil
EC Number:
810-543-2
Cas Number:
1618093-67-6
Molecular formula:
n/a
IUPAC Name:
Amides, from diethylenetriamine and hydrogenated palm oil
Test material form:
solid: compact
Details on test material:
- Name of test material (as cited in study report): Amidoamine (UVCB)
- Substance type: Amidoamine
- Chemical name: Glycerides, C14-20, reaction products with diethylenetriamine (preliminary naming)/Amides, from diethylenetriamine and hydrogenated palm oil
- CAS 85409-11-6/1618093-67-6
- Physical state: pale yellowish solid at 20 °C
- Batch No.: K8 4309 L481
- Expiry date of batch: 09 March 2017
- Purity: 100 % (UVCB)
- Storage condition of test material: Room temperature, protected from light
- Stability: stable under test conditions
Specific details on test material used for the study:
Test item without emulsifier was investigated.

Amidoamine (UVCB)
Pulcra ID: DE07_2014_012_BEL66 (amidoamine without emulsifier)
Physical state: pale yellowish solid at 20 °C
Batch No.: K8 4309 L481
Expiry date of batch: 09 March 2018
Purity: 100 % (UVCB)
Stability: stable under test conditions
Storage condition of test material: Room temperature, protected from light

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
Eyes from adult cattle (typically 12 to 60 months old) were obtained from a local abattoir as a by-product from freshly slaughtered animals. The eyes were excised by an abattoir employee after slaughter, and were placed in Hanks’ Balanced Salt Solution (HBSS) supplemented with antibiotics (penicillin at 100 IU/mL and streptomycin at 100 µg/mL). They were transported to the test facility over ice packs on the same day of slaughter. The corneas were prepared
immediately on arrival.

Test system

Vehicle:
other: 20% w/v solution in 0.9% w/v sodium chloride solution
Controls:
yes, concurrent vehicle
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
the test item was ground to a powder and prepared as a 20% w/v solution in 0.9% w/v sodium chloride solution. 0.75 mL of the test item preparation or control items were applied to the appropriate corneas.
Duration of treatment / exposure:
Each cornea holder was incubated, anterior chamber uppermost, at 32 ± 1 ºC for 240 minutes. At the end of the exposure period the test item and control items were removed from the anterior chamber and the cornea was rinsed three times with fresh complete MEM containing phenol red before a final rinse with complete MEM without phenol red. The anterior chamber was refilled with fresh complete MEM without phenol red.
A post-treatment opacity reading was taken and each cornea was visually observed.
Following the opacity measurement the permeability of the corneas to sodium fluorescein was evaluated. The medium from the anterior chamber was removed and replaced with 1 mL of sodium fluorescein solution (5 mg/mL). The dosing holes were plugged and the holders incubated, anterior chamber uppermost, at 32 ± 1 ºC for 90 minutes.
Number of animals or in vitro replicates:
Three corneas with opacity values close to the median value of all corneas were allocated to the negative control. Three corneas were also allocated to the test item and three corneas to the positive control item.
Details on study design:
All eyes were macroscopically examined before and after dissection. Only corneas free of damage were used.

The cornea from each selected eye was removed leaving a 2 to 3 mm rim of sclera to facilitate handling. The iris and lens were peeled away from the cornea. The isolated corneas were immersed in a dish containing HBSS until they were mounted in Bovine Corneal Opacity and Permeability (BCOP) holders.

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Run / experiment:
240 minutes incubation
Value:
0.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
In Vitro Irritancy Score: 1.8
Positive controls validity:
valid
Remarks:
In Vitro Irritancy Score: 68.8
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
The positive control In Vitro Irritancy Score was within the range of 73.8 – 103.6. The positive control acceptance criterion was therefore satisfied.

The negative control gave opacity of ≤4.7 and permeability ≤0.085. The negative control acceptance criterion was therefore satisfied.

The corneas treated with the test item were clear post treatment.

The corneas treated with the negative control item were clear post treatment.

The corneas treated with the positive control item were cloudy post treatment.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
No category. Not requiring classification to UN GHS or EU CLP.
Executive summary:

In a Klimisch 1 GLP study from Warren (2016) the potential of the test item to induce serious eye damage and to identify test items not requiring classification for eye irritation or serious eye damage using the Bovine Corneal Opacity and Permeability (BCOP) test method was investigated. The test item's In Vitro Irritancy Score was < 3. The test item was not considered to induce serious eye damage and is not requiring classification to UN GHS or EU CLP.

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