(3R,4S,5R)-3,4,5-trihydroxycyclohex-1-enecarboxylic acid

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

other information

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

According to: OECD Guideline for the testing of Chemicals 439: In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method (Original Guideline adopted July 26, 2013), and as described in detail in the Protocol for: In Vitro EpiDermTM Skin Irritation Test (EPI-200-SIT) for use with MatTek Corporation’s Reconstructed Human Epidermal Model EpiDerm (EPI-200), Rev. 1/19/2010

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

(-)-Shikimic acid

IUPAC Name:

(-)-Shikimic acid

Details on test material:

Identification: (-)-Shikimic acid
CAS No.: 138-59-0
Roche No.: Ro0642565-000
Batch: 1329R037/ Lot. No. 0700791872
Purity: Min. 98.0% (dose calculation was not adjusted to purity)
Expiry Date: 22 August 2014
Storage Conditions: At room temperature, protected from light*
Stability in Solvent: Not indicated by the Sponsor

Test system

Amount / concentration applied:

Each approximately 25 mg of the neat test item were applied to three skin equivalents. Additionally the test item treated skin equivalents were wetted with 25 µL DPBS per tissue.

Duration of treatment / exposure:

The skin equivalents were exposed to the neat test item for on hour.

Details on study design:

Each approximately 25 mg of the neat test item and 30 µL of the positive control (5% SDS in deionised water) or the negative control (DPBS) were applied to each three skin equivalents per dose group. Additionally the test item treated skin equivalents were wetted with 25 µL DPBS per tissue. The skin equivalenst were exposed to the neat test item and the controls for 1 hour. Subsequently the test item as well as the controls were rinsed off the skin equivalents and they were further incubated for about 41 hours. Afterwatrds the viability was determined using the MTT assay.

Results and discussion

In vivo

Irritant / corrosive response data:

Compared to the relative absorbance value of the negative control the mean relative absorbance value was not reduced (100.8%) after exposure of the test item to the skin tissues. The threshold for irritancy is ≤ 50%. Therefore, the test item is not considered to possess an irritant potential.

Any other information on results incl. tables

Results after treatment with (-)-Shikimic acid and the controls

Dose Group	Treat-ment Interval	Absorbance 570 nm Tissue 1*	Absorbance 570 nm Tissue 2*	Absorbance 570 nm Tissue 3*	Mean Absorbance of 3 Tissues	Rel. Absorbance [%] Tissue 1, 2 + 3**	Relative Standard Deviation [%]	Mean Rel. Absorbance [% of Negative Control]***
Negative Control	60 min	1.666	1.624	1.606	1.632	102.1 99.5 98.4	1.9	100.0
Positive Control	60 min	0.063	0.065	0.062	0.063	3.8 4.0 3.8	2.7	3.9
Test Item	60 min	1.592	1.599	1.744	1.645	97.5 98.0 106.8	5.2	100.8

 

*      Mean of three replicate wells after blank correction
**       relative absorbance per tissue [rounded values]:
***     relative absorbance per treatment group [rounded values]:

 

Optical evaluation of the MTT-reducing capacity of the test item after 1 hour incubation with MTT-reagent did not show blue colour.

The mean relative absorbance value of the test item, corresponding to the cell viability, did not decrease (100.8%; threshold for irritancy:≤50%), consequently the test item was non irritant to skin. 

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

In conclusion, it can be stated that in this study and under the experimental conditions reported, (-)-Shikimic acid is not irritant to skin.

Executive summary:

This in vitro study was performed to assess the irritation potential of (-)-Shikimic acid by means of the Human Skin Model Test.

Each three tissues of the human skin model EpiDerm^™were treated with the test item, the negative or the positive control for 60 minutes.

Each about 25 mg of the test item was applied to each tissue.

30 µL of either the negative control (DPBS) or the positive control (5% SLS) were applied to each tissue.

The test item and the positive and negative controls were washed off the skin tissues after 60 minutes treatment. After further incubation for about 41 hours the tissues were treated with the MTT solution for 3 hours following approximately 70 hours extraction of the colorant from the cells. The amount of extracted colorant was determined photometrically at 570 nm.

After treatment with the negative control the absorbance values were well within the required acceptability criterion of mean OD equal or greater than 0.8 and less or equal than 2.8 for the 60 minutes treatment interval thus showing the quality of the tissues.

Treatment with the positive control induced a decrease in the relative absorbance compared to the negative control to 3.9% thus ensuring the validity of the test system.

The rel. standard deviations between the % variabilities of the test item, the positive and negative controls were below 6% (threshold of the "OECD Guideline for the Testing of Chemicals 439: In vitro Skin Irritation: Reconstructed Human Epidermis Test Method”: 18%), thus ensuring the validity of the study.

Compared to the relative absorbance value of the negative controlthe mean relative absorbance value was not reduced (100.8%) after exposure of the test item to the skin tissues. The threshold for irritancy is ≤ 50%. Therefore, the test item is not considered to possess an irritant potential.