Cashew (Anacardium occidentale) Nutshell Extract, Decarboxylated, Distilled, oligomerisation products with 1-chloro-2,3-epoxypropane

Endpoint summary

Description of key information

- Skin irritation / corrosion: not irritating (based on the results of a reliable OECD 439 in vitro study; GLP)

- Eye irritation: not irritating (based on the results of two reliable in vitro studies: OECD 437 (BCOP) and a Human Cornea Model (similar to OECD 492))

- Respiratory irritation: no study available

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin 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:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: not applicable (Human reconstitued skin)

Strain:

other: not applicable

Details on test animals or test system and environmental conditions:

Commercially available Epi-200-SIT-Kit was used.
The EpiDermTM tissue consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multi-layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues are cultured on specially prepared cell cultures inserts.

Epi-200 tissues were procured from MatTek In Vitro Life Science Laboratories, Bratislava.
Day of delivery: 11. Dec. 2012; batch: 16865.

Type of coverage:

open

Preparation of test site:

other: not applicable

Vehicle:

unchanged (no vehicle)

Controls:

other: not applicable

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 µl/well

Duration of treatment / exposure:

35 - 60 min

Observation period:

not applicable

Number of animals:

three tisue were used

Irritation / corrosion parameter:

other: other: cell viability

Value:

90

Remarks on result:

other: Cell viability 90% (above threshold)

% Formazan Production

Designation	CARDOLITE NC-513	Positive Control
% Formazan production (Tissue 1)	92.3%	6.7%
% Formazan production (Tissue 2)	88.8%	6.8%
% Formazan production (Tissue 3)	89.1%	7.2%
% Formazan production Mean	90.0%	6.9%

Interpretation of results:

other: Not irritating according to EU GHS criteria

Conclusions:

In a GLP-study according to OECD Test Guideline 439, the registered substance was not skin irritating.

Executive summary:

The Skin Irritation Potential of CARDOLITE NC-513 was determined in the Human Skin Model Test following EU-Method B.46 resp. OECD 439.

One valid experiment was performed. Three tissues of the human skin model EpiDerm^TMwere treated withCARDOLITE NC-513for 60 minutes. 30 µL of the liquid test item (using a nylon mesh) were applied to each tissue and spread to match the tissue size (0.63 cm²; as indicated by supplier). DPBS-buffer was used as negative control, 5 % SDS-solution was used as positive control.

After treatment with the negative control, the absorbance values were within the required acceptability criterion of 1.0 < mean OD < 2.5. The positive control showed clear irritating effects. Variation within tissues was acceptable (< 18 %).

After the treatment with the test item, the relative absorbance values were reduced to 90.0 %. This value is well above the threshold for irritation potential (50 %). Therefore, CARDOLITE NC-513 is considered as not irritant in the Human Skin Model Test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

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:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

not applicable

Vehicle:

other: 0.9% sodium chloride solution

Controls:

other: not applicable

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µl/per eye
- Concentration (if solution): 20%

Duration of treatment / exposure:

10 minutes

Observation period (in vivo):

not applicable

Number of animals or in vitro replicates:

not applicable

Details on study design:

see below

Irritation parameter:

in vitro irritation score

Value:

0.002

 

IVIS – In Vitro Irritancy Score

Test Group	IVIS	Mean IVIS	Relative Standard Deviation IVIS
Negative Control 0.9 % NaCl	0.420	0.380	14.8%
	0.405
	0.316
Positive Control 10 % NaOH	162.467	149.960	26.0%
	106.209
	181.205
Solvent Control (olive oil )	0.555	0.533	10.4%
	0.574
	0.469
CARDOLITE NC-513	-0.010	0.002	2514.7%
	-0.042
	0.057

Interpretation of results:

other: EU GHS criteria not met

Conclusions:

In an in vitro eye irritation test according to OECD Test Guideline 437 (BCOP), the test item CARDOLITE NC-513 showed no severe eye irritation.

Executive summary:

In an in vitro eye irritation test according to OECD Test Guideline 437 (BCOP), the corneal irritation and damage potential of CARDOLITE NC-513

was investigated by quantitative measurements of changes in opacity and permeability in a bovine cornea.One experiment were performed. Bovine corneas were used. They were collected from slaughtered cattle which were between 12 and 60 months old.

The test item CARDOLITE NC-513 was dissolved in olive oil (10 % solution) and brought onto the cornea of a bovine eye which had been incubated with cMEM without phenol red at 32 ± 1 °C for one hour and whose opacity had been measured. The test item was incubated on the cornea for 10 minutes at 32 ± 1 °C. After removal of the test item and two hours post-incubation, opacity and permeability values were measured.

Olive oil was used as negative control. The negative control showed no irritating effect on the cornea.

10% sodium hydroxide solution was used as positive control. The positive control induced a severe irritation on the cornea.

The test item CARDOLITE NC-513 showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is 0.002.

In conclusion, it can be stated that in this study and under the experimental conditions reported, the test item CARDOLITE NC-513 possesses no eye irritation potential (according to ICCVAM).

According to OECD Guideline no. 437 (2009), a substance that induces an IVIS ≥ 55.1 is defined as a corrosive or severe irritant.

Reference 2

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:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Version / remarks:

OECD Guideline for the Testing of Chemicals, Part 492, adopted 18. Jun. 2019, “Reconstructed Human Cornea-like Epithelium (RhCE) test method for identifying chemicals not requiring classification and labelling for eye Irritation or serious eye damage”

Deviations:

not applicable

Principles of method if other than guideline:

This study was performed in order to evaluate the potential of CARDOLITE NC-513 to evoke eye irritation in a human cornea model in an in-vitro study.
The eye irritation test refers to the production of irreversible tissue damage following the application of a test material on a human cornea model.
Irritating materials are identified by their ability to produce a decrease in cell viability as determined, by using the MTT reduction assay below defined threshold levels at specified exposure periods.
The principle of the human cornea model assay is based on the hypothesis, that irritating chemicals are able to penetrate the epithelium by diffusion or erosion, and are cytotoxic to the underlying cell layers.

GLP compliance:

yes (incl. QA statement)

Species:

other: not applicable

Strain:

other: not applicable

Details on test animals or tissues and environmental conditions:

not applicable

Vehicle:

unchanged (no vehicle)

Controls:

other: not applicable

Amount / concentration applied:

100µl/tissue

Duration of treatment / exposure:

3, 30, 60 min

Observation period (in vivo):

not applicable

Number of animals or in vitro replicates:

not applicable

Irritation parameter:

other: ET50

Value:

107.9

Remarks on result:

other: 107.9% at 3mins

Irritation parameter:

other: ET50

Value:

96.6

Remarks on result:

other: 96.6% at 30 mins

Irritation parameter:

other: ET50

Value:

105.4

Remarks on result:

other: 105.4% at 60 mins

Interpretation of results:

other: EU GHS criteria not met

Conclusions:

Using a Human Cornea Model following MTT ET-50 Protocol, the Eye Irritation Potential of CARDOLITE NC-513 was determined as not irritating.

Executive summary:

The Eye Irritation Potential of CARDOLITE NC-513 was determined using a Human Cornea Model following
MTT ET-50 Protocol. One valid experiment was performed.

The test item CARDOLITE NC-513 was applied to a three-dimensional human cornea model tissue for three different exposure times in duplicate (3 min., 30 min. and 60 min.).

100 µL of the liquid test item were applied to each tissue.

Deionised water was used as negative control, 0.3% Triton X 100 solution was used as positive control.

After treatment, the respective substance was rinsed from the tissue; then, cell viability of the tissues was evaluated by addition of MTT which can be reduced to a blue formazan. Formazan production was measured by measuring the optical density (OD) of the resulting solution.

After treatment with the negative control, the absorbance values were well within the historical range and showed no cytotoxic effects. The positive control showed clear irritating effects.

After 3 minutes treatment with the test item, the relative absorbance values increased to 107.9 %. After 30 minutes treatment, relative absorbance values were reduced to 96.6 %. And after 60 minutes treatment, the relative absorbance values increased to 105.4 %. These values lie within the range of the negative control.

From these values, it is assumed that the ET₅₀of the test item CARDOLITE NC-513 is > 60 min.

Therefore, the test item CARDOLITE NC-513 is considered as not/minimal eye irritant in the Human Cornea Model test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

- Skin / Eye Irritation in vitro

The registered substance did not induce skin irritation in vitro. Therefore, it can be concluded that it is also not skin corrosive.

The substance was not severly eye irritating according to the BCOP. Subsequently, using a Human Cornea model, it was shwon to be non eye irritating.

 

- Skin / Eye Irritation in vivo

In vivo irritation studies do not need to be conducted because the available in vitro data are adequate for classification and risk assessment.

Justification for classification or non-classification

Based on results of reliable in vitro studies, the substance does not meet the CLP classification criteria with respect to irritation or corrosion to the skin and eye, as set out in Regulation (EC) No. 1272/2008.