Indolo[2,3-a]quinolizine-1-propanoic acid, 1-ethyl-1,2,3,4,6,7,12,12b-octahydro-.alpha.-(hydroxyimino)-, ethyl ester, (1S,12bS)-

Skin irritation / corrosion

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

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)

Test material

Test material information

Constituent 1

Reference substance name:

/-/-1α-Ethyl-1,2,3,4,6,7,12,12b α-octahydro-indolo[2,3-a]-quinolizine-1β-yl-ethyl-pyruvate oxime

EC Number:

617-734-8

Cas Number:

85647-38-7

Molecular formula:

C22H29N3O3

IUPAC Name:

/-/-1α-Ethyl-1,2,3,4,6,7,12,12b α-octahydro-indolo[2,3-a]-quinolizine-1β-yl-ethyl-pyruvate oxime

Test material form:

solid: crystalline

Details on test material:

Test item: hydroxy-imino-ethyl-esterBatch No.: K52212NPhysical state: solid substance, powderColour: light-yellowStorage: at 15-30 °C

Test system

Details on study design:

Human skin:EpiSkin(TM) Small Model (EpiSkin(TM)SM), manufactured by EPISKIN SNC Lyon, France, is a three-dimensional human epidermis model. Adult human-derived epidermal keratinocytes are seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. A highly differentiated and stratified epidermis model is obtained after 13-day culture period comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum (Tinois et al., 1994). Its use for skin irritation testing involves topical application of test materials to the surface of the epidermis, and the subsequent assessment of their effects on cell viability.Justification for selection of the test system:The EPISKIN model has been validated for irritation testing in an international trial.

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Value:

50

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

Disks of EPISKIN (three units / chemical) were treated with test item and incubated for 15 minutes at room temperature. Exposure of test material was terminated by rinsing with PBS 1x solution. Epidermis units were then incubated at 37 °C for 42 hours in an incubator with 5% CO2. The viability of each disk was assessed by incubating the tissues for 3 hours with MTT solution at 37°C in 5% CO2 protected from light. The precipitated formazan was then extracted using acidified isopropanol and quantified spectrophotometrically.SDS (5% aq.) and PBS treated epidermis were used as positive and negative controls respectively. For each treated tissue viability was expressed as a percentage relative to negative control.The test item is considered to be irritant to skin, if the mean relative viability after 15 minutes exposure and 42 hours post incubation is less or equal (≤) to 50% of the negative control.In this in vitro skin irritation test using the EPISKIN model, the test item showed significantly reduced cell viability in comparison to the negative control. The mean relative viability of the test item treated tissues was determined to be 50 %. Accordingly, the test item was considered as irritant to skin.Positive and negative controls showed the expected cell viability values within acceptable limits. The experiment was considered to be valid.

Any other information on results incl. tables

Substance	Optical Density (OD)		Viability(%)
Negative Control: 1 x PBS	mean	0.836	100
	standard deviation (SD)		8.44
Positive Control: SDS (5%aq.)	mean	0.033	4
	standard deviation (SD)		1.83
Test Item: Hydroxy-imino-ethyl-ester	mean	0.419	50
	standard deviation (SD)		12.00

Applicant's summary and conclusion

Interpretation of results:

irritating

Remarks:

Migrated informationIrritant to skin (UN GHS Category 2).Criteria used for interpretation of results: OECD GHS

Conclusions:

The results obtained from this in vitro skin irritation test, using the EPISKIN model, indicated that the test item reveals skin irritation potential under the utilised testing conditions. The test item HYDROXY-IMINO-ETHYL-ESTER (CAS No. 85647-38-7) is considered to be irritant to skin (UN GHS Category 2).