Glycine max. (L.) Merr. (Fabacee) aqueous alcoholic ext., concentrate by sequential extractions and filtration

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May 24 - June 03, 2013

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: L’Oréal Standard Operating Procedure: “EpiSkin™ Skin Irritation Test Method“; - ECVAM Skin Irritation Validation Study - Validation of the EpiSkin™ Test Method15 min - 42 hours for the Prediction of Acute Skin Irritation of Chemicals. Version 1.8, 02-2009

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: ECVAM Performance Standards for in vitro Skin Irritation Test Methods based on Reconstructed human Epidermis (RhE). Updated Version 24-Aug-2009

Test guideline 5

Qualifier:

according to guideline

Guideline:

other: ISO 10993-10, Annex D: 2010, “Biological evaluation of medical devices - Part 10: Tests for irritation and skin sensitization”

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Glycine max. (L.) Merr. (Fabacee) aqueous alcoholic ext., concentrate by sequential extractions and filtration

EC Number:

938-980-5

IUPAC Name:

Glycine max. (L.) Merr. (Fabacee) aqueous alcoholic ext., concentrate by sequential extractions and filtration

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Name: Soybean (Glycine max) dry purified extract
Batch No.: 30432/M1
Physical State: powder
Colour: yellow to light brown
Purity: 100%
Storage Conditions: at room temperature

Test system

Details on study design:

Cytotoxicity is expessed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 15 min exposure and 42 h post incubation period and compared to those of the concurrent negative controls.

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

other: other: skin irritant potential

Remarks on result:

other:

Remarks:

Remarks: no irritant effects. (migrated information)

Any other information on results incl. tables

Pre-Experiments

The mixture of 10 mg test item per 2 mL MTT medium showed reduction of MTT compared to the solvent. The mixture turned blue/purple. For quantitative correction of results, the part of absorption due to the non specific reduction of MTT (NSMTT) was determined by using killed tissues (4). Therefore, three tissues were treated with the test item (KT) and with PBS (KU), respectively. NSMTT was calculated relative to the negative control of living tissues (NK) according to the following formula:

NSMTT = [(ODKT - ODKU)/ODNK] * 100

Mean ODKT = 0.130

Mean ODKU = 0.150

Mean ODNK = 0.788

Since the calculated NSMTT was < 0% (-2.5%) the true MTT metabolic conversion (TODTT) of the test item treated living tissues (TM) was not calculated. The mixture of 10 mg of the test item per 90 μl aqua dest. showed no relevant colouring detectable by unaided eye-assessment.

Experiment

Name	NegativeControl			PositiveControl			TestItem
Tissue	1	2	3	1	2	3	1	2 a	3
absoluteOD550	0.798	0.832	0.885	0.218	0.330	0.247	0.831	0.467	0.865
	0.761	0.842	0.869	0.213	0.333	0.242	0.841	0.455	0.840
OD550(blank- corrected)	0.755	0.788	0.842	0.175	0.287	0.203	0.787		0.821
	0.718	0.798	0.825	0.169	0.290	0.198	0.798		0.797
meanOD550oftheduplicates(blank- corrected)	0.736	0.793	0.833	0.172	0.288	0.201	0.792		0.809
totalmeanOD550of3replicatetissues(blank- corrected)	0.788*			0.220			0.801
SDOD550	0.046			0.054			0.014
relativetissue viabilities[%]	93.5	100.7	105.8	21.8	36.6	25.5	100.6		102.7
mean relative tissue viability [%]	100.0			28.0**			101.7
SDtissueviability[%]***	6.2			7.7			1.5
CV[%viability]	6.2			27.5			1.5

a Tissue 2 was classified as Outlier acc. to Nalimov, Grubbs and Dixon and not used for evaluation * Corrected mean OD550 of the negative control corresponds to 100% absolute tissue viability. ** Mean relative tissue viability of the three positive control tissues is <= 40%. *** The standard deviation (SD) obtained from the three concurrently tested tissues is < 18%

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item showed no irritant effects. The relative mean tissue viability after 15 min of exposure and 42 h post incubation was >50%. The test item is therefore classified as “non-irritant”.