Lupine, L. albus, ext.

Endpoint summary

Description of key information

The test item was tested for eye irritation by an alternative method (application to fibroblast of rabbit cornea and cytotoxicity assessment by the neutral red release method). The substance has insignificant cytotoxicity.

The product applied as such in a single patch test, can be considered as non-irritant after an application with the help of an occlusive patch for 48 h on 11 volunteers.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Reason / purpose for cross-reference:

data waiving: supporting information

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20-21 January 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Remarks:

Study performed according to French offical method published in December 1999 in National register N° 302, and Good Laboratory Practice

Qualifier:

according to guideline

Guideline:

other: Appendix VI of the French National Register N°302 of December, 1999

Deviations:

not specified

Principles of method if other than guideline:

This test is an alternative method which aims to assess a test item eye irritant potential,. The principle is based on the test item cytotoxicity assessment by determination of the concentration which leads to 50% of cells death (IC50) on a cell monolayer, using the Neutral Red Uptake Method.

GLP compliance:

yes

Species:

rabbit

Strain:

other: Rabbit cornea fibroblasts: SIRC line (ATCC CCL60)

Details on test animals or tissues and environmental conditions:

Rabbit cornea fibroblasts: SIRC line (ATCC CCL60)

Vehicle:

other: physiological serum or cottonseed oil

Controls:

yes, concurrent vehicle

yes, concurrent positive control

Amount / concentration applied:

Dilution at 25% and 50% of the test material are used.

Duration of treatment / exposure:

contact with the test item = 60 secondes

Observation period (in vivo):

Not applicable

Duration of post- treatment incubation (in vitro):

Not specified

Number of animals or in vitro replicates:

Each sample and positive control are tested on three cultures wells by assessed concentration. Negative control is tested on three culture wells per plate.,

Irritation parameter:

other: Percentage of mortality observed at the 50% dilution

Value:

3

Vehicle controls validity:

not applicable

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

other: Estimated IC 50 (%)

Value:

> 50

Vehicle controls validity:

not applicable

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Substance tested	Dose (%)	mean percent of cells mortality
SDS	0.2%	85
SDS	0.05%	39
SDS	0.01%	20
test item	25%	10
test item	50%	3

Interpretation of results:

other: insignificant toxicity

Remarks:

according to table of French national register N°302, December 1999

Conclusions:

The cytotoxicity of tested material is insignificant.
IC 50 is greater to 50% and the percentage of mortality observed at the dilution 50% is 3%.
Under the retained experimental conditions, the cytotoxicity of the tes item may be classified as insignificant cytotoxicity according to the adopted scale

Executive summary:

An in vitro eye irritation study was performed on rabbit cornea fibroblasts according to French national Method published in December 1999 in National register N° 302. The principle of the method is based on assessing the cytotoxicity of the product tested by identifying the concentration causing 50% mortality (IC50) using the technique of neutral red release.

Positivie and Negative (vehicle) controls were used.

The IC50 was up to 50% and the percentage of mortality at the dilution 50% was 3%.

According to this method the cytotoxicity is considered as insignificant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

An in vitro eye irritation study was performed, on rabbit cornea fibroblasts, according to French national Method published in December 1999 in National register N° 302.

The principle of the method is based on assessing the cytotoxicity of the product tested by identifying the concentration causing 50% mortality (IC50) using the technique of neutral red release.

The IC50 was up to 50% and the percentage of mortality at the dilution 50% is 3%.

According to the method the cytotoxicity is considered as insignificant.

Skin irritation was assessed by a single patch test, on 11 volunteers (men and women, aged from 18 to 65 years) without any dermatological lesion on the experimental area. 0.02 mL of test item was applied, on the external face of the arm, maintained for 48 hours in contact with the skin, with the help of an occlusive patch (Finn Chamber).

The average irritation index was 0.

Under the conditions employed, in this study, the test item was considered as non irritant for skin.

Justification for classification or non-classification

On a single patch test, performed on 11 volunteers, the product has been demonstrated to be non irritant for skin.

On an in vitro test, based on neutral red release method on rabbit cornea fibroblasts, the test item has been considered to have insignificant cytotoxicity.

Also, considering the composition of the registered substance, mainly composed of proteins from Lupinus Albus seeds, the registered substance does not need to be classified as irritating for eyes and skin under CLP Regulation (1272/2008).