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Administrative data

Description of key information

Skin corrosion/irritation:

Key study. Test method according to OECD 431 and EU B.40 Bis. GLP study. The substance is not corrosive or irritant to reconstructed human epidermis model based on cell viability.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation / corrosion
Remarks:
in vitro
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: B.40 Bis In vitro skin corrosion: Human skin model test
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Deviations:
no
GLP compliance:
yes
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: Reconstructed Human Epidermis
Source strain:
other: MatTek´s EpiDerm System
Details on animal used as source of test system:
Reconstructed Human Epidermis.
Justification for test system used:
In vitro validated system.
Vehicle:
water
Details on test system:
The Epiderm Skin Corrosivity/Irritation Test (Model EPI-200) was performed to reveal possible irreversible tissue damages of the skin following the application of the test substance. Firstly the test substance was topically applied for 3 minutes and 1 hour to the epidermal surfaces of three-dimensional human epidermis models, followed by immediate determination of the cytotoxic effect. As there was no corrosive effect observed, the test substance was topically applied for 15 minutes to the epidermal surfaces of three-dimensional human epidermis models. After a post-incubation of 42 hours, a cell viability test was performed.

Test system
MatTek´s EpiDerm System consists of normal, human-derived epidermal keratinocytes which have been cultured form a multilayered, 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 EpiDerm™ tissues (surface 0.6 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs, 10 mm diameter) and shipped as kits, containing 24 tissues on shipping agarose.
Control samples:
not required
Amount/concentration applied:
Application of the reference substances:
50 µL of each reference substance were dispensed directly atop the Epi-200 tissue.

Application of the test substance:
The application spoon was filled with 25 mg finely grounded test substance. The "spoonful" was levelled by gently scratching the excess material away, avoiding compression.
25 µL H2O were added for wetting of the test substance.
Duration of treatment / exposure:
Epiderm Skin Corrosivity Test: Exposure times were 3 minutes and 1 hour.
Epiderm Skin Irritation Test: Exposure time was 15 minutes.
Duration of post-treatment incubation (if applicable):
42 h.
Number of replicates:
Two tissue replicates were used for each treatment (exposure time) in the corrosivity test.
Three tissue replicates were used for each treatment (exposure time) in the irritation test.
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Corrosivity test
Value:
101.3
Remarks on result:
other: 3 min exposure.
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Corrosivity test
Value:
125.5
Remarks on result:
other: 1h exposure
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Irritation test
Value:
97.93
Remarks on result:
other: 3 min exposure
Irritant / corrosive response data:
Corrosivity test:
Mean tissue viability = 101.3 % (3 min exposure).
Mean tissue viability = 125.5 % (1 h exposure).

Irritation test:
Mean tissue viability = 97.93 % (3 min exposure).

Epiderm Skin Corrosivity Test:

Assay acceptance criteria according to the protocol INVITTOX n°119 by ECVAM:

- The mean optical density (OD) of the tissues, treated with deionised water (negative control) was 2.560 after 3 minutes, and 1.936 after 1 hour of exposure, that is higher than 0.8, as required by the assay acceptance criteria.

- The mean tissue viability of the 3 minutes positive control was 14.9 %, that is lower than 30 %, as required by the assay acceptance criteria.

- The maximum inter tissue viability differences of the test substance treated skin discs were 10.6 % for 3 minutes and 0.1 % for 1 hour exposure, that is below 30 % as required by the assay acceptance criteria.

Test substance:

The mean percentage viability of the treated skin discs after 3 minutes of exposure was 101.3 % which is above the threshold of 50 % for classification.

The mean percentage viability of the treated skin discs after 1 hour of exposure was 125.5 % which is above the threshold of 15 % for classification.

Epiderm Skin Irritation Test:

Assay acceptance criteria according to the protocol used during the ECVAM validation study:

- The mean OD of the tissues, treated with deionised water (negative control) was 1.799 , that is higher than 1.0, as required by the assay acceptance criteria.

- The mean tissue viability of the positive control was 6.88 %, that is lower than 20 %, as required by the assay acceptance criteria.

- The standard deviation calculated from individual percentual tissue viabilities of the test substance treated skin discs was 4.42, that is below 11 as required by the assay acceptance criteria.

Test substance:

The mean percentage viability of the treated skin discs was 97.93 % which is above the threshold of 50 % for classification.

According to the results of this study and the Directive 2001/59/EC, the test substance is considered to be non-corrosive and non-irritant to skin

Interpretation of results:
GHS criteria not met
Remarks:
CLP Implementation.
Conclusions:
Conclusion
According to the results of this study, the test substance is considered to be non-corrosive and non-irritant to skin.
Executive summary:

The Epiderm Skin Corrosivity/Irritation Test (Model EPI-200) was performed to reveal possible irreversible tissue damages of the skin following the application of "TAZOBACTAM ACID".

Firstly the test substance was topically applied for 3 minutes and 1 hour to the epidermal surfaces of three-dimensional human epidermis models, followed by immediate determination of the cytotoxic effect. As there was no corrosive effect observed, the test substance was topically applied for 15 minutes to the epidermal surfaces of three-dimensional human epidermis models. After a post-incubation of 42 hours, a cell viability test was performed.

Investigations performed were in conformance with the Directive 2000/33/EC, B. 40, "Skin corrosion", the OECD-Guideline 431, "In Vitro Skin Corrosion: Human Skin Model Test ", 13 April 2004 and the ESAC statement, 27 April 2007.

Results:

Corrosivity test:

Mean tissue viability = 101.3 % (3 min exposure).

Mean tissue viability = 125.5 % (1 h exposure).

 

Irritation test:

Mean tissue viability = 97.93 % (3 min exposure).

 

According to the results of this study , the test substance is considered to be non-corrosive and non-irritant to skin.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Based on the available information, the substance is not classified for skin irritation/corrosion according to CLP Regulation (EC) No 1272/2008.