Fatty acids, vegetable-oil, sulfated, sodium salts

Endpoint summary

Description of key information

Not irritant for skin, slightly irritant for eyes

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 13-19, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study according to well-documented and supported procedures for which no guideline (OECD, etc.) exists

Justification for data waiving:

other:

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline available

Principles of method if other than guideline:

The principle of the assay was based on the measurement of cytotoxicity in reconstituted human epidermal cultures following topical exposure to the test material for 15 minutes and incubation for 42 hours by means of the colourmetric MTT reduction assay.

GLP compliance:

yes (incl. QA statement)

Species:

other: reconstituted human epidermis model

Details on test animals or test system and environmental conditions:

TEST ANIMALS - EPISKIN Model Kit
- Source: SkinEthic Laboratories, Nice, France
- Date received: 13 April 2010
- All other template details: Not applicable

ENVIRONMENTAL CONDITIONS: Not applicable

IN-LIFE DATES: Not applicable

Type of coverage:

other: uniform topical coverage

Preparation of test site:

other: Not applicable

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 10 µl

VEHICLE: Not applicable

Duration of treatment / exposure:

15 minutes

Observation period:

Not applicable

Number of animals:

Three tissues per test material, positive control, and negative control for a total of 9 skin models analyzed

Details on study design:

TEST SITE
- Area of exposure: entire EPISKIN tissue
- % coverage: 100
- Type of wrap if used: Not applicable

REMOVAL OF TEST SUBSTANCE
- Washing: rinsed with dulbeccos phosphate buffered saline (PBS) with calcium and magnesium
- Time after start of exposure: at the end of the 15 minute exposure period

SCORING SYSTEM: optical density at 540 nm was measured and divided by the negative control optical density and multiplied by 100 to obtain a percentage for the relative mean viability.
- If the relative mean tissue viability was less or equal to 50%, the compound is predicted to be an irritant.
- If the relative mean tissue viability was greater than 50%, the compound is predicted not to be an irritant.

Irritation / corrosion parameter:

other: other: relative mean viability based on OD540 measurements

Value:

102.2

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 15 minutes. Max. score: 108.9. Reversibility: other: not applicable. Remarks: scores are percentages of mean negative control OD540 values . (migrated information)

Irritant / corrosive response data:

See Table 1 for raw data values

Other effects:

Test material was shown to directly reduce MTT. However, rinsing was effective at removing the test material on or in the tissue following each exposure period and no degree of interference was noted.

Table 1. Individual and Mean OD540 Values and Percentage Viabilities for the Negative Control Material, Positive Control Material and Test Material.

Material	OD540of tissues	Mean OD540of triplicate tissues (±SD)	Relative individual tissue viability (%)	Relative mean viability (%)	± SD of Relative mean viability (%)
Negative control material	0.638	0.629 (0.008)	101.4	100	1.2
	0.623		99.0
	0.627		99.7
Positive control material	0.194	0.175 (0.016)	30.8	27.9	2.6
	0.164		26.1
	0.168		26.7
Test material	0.623	0.643 (0.036)	99.0	102.2	5.8
	0.621		98.7
	0.685		108.9

 

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test material was considered to be a non-irritant to the skin.

Executive summary:

Summary from data report:

Introduction: The purpose of this test was to evaluate the skin irritation potential of the test material using the EPISKIN^TMreconstituted human epidermis model after a treatment period of 15 minutes followed by a post-exposure incubation period of 42 hours. The principle of the assay was based on the measurement of cytotoxicity in reconstituted human epidermal cultures following topical exposure to the test material by means of the colourmetric MTT reduction assay. Cell viability is measured by enzymatic reduction of yellow MTT tetrazolium salt (3 -[4,5 -dimethylthiazol-2 -yl]-2,5 -diphenyl-tetrazolium bromide) to a blue formazan salt (within the mitochondria of viable cells) in the test material treated tissues relative to the negative controls. The concentration of the inflammatory mediator IL-1alpha in the culture medium retained following the 42 hour post-exposure incubation period is also determined for test materials which are found to be borderline non-irritant based upon the MTT reduction endpoint. This complimentary end-point will be used to either confirm a non-irritant result or will be used to override the non-irritant result.

 

Methods: Triplicate tissues were treated with the test material for an exposure period of 15 minutes. At the end of the exposure period each tissue was rinsed before incubation for approximately 42 hours. At the end of the post-exposure incubation period each tissue was taken for MTT-loading. The maintenance medium from beneath each tissue was transferred to pre-labelled micro tubes and stored in a freezer for possible inflammatory mediator determination. After MTT loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals out of the MTT loaded tissues.

 

At the end of the formazan extraction period each tube was mixed thoroughly and duplicate 200µl samples were transferred to the appropriate wells of a pre-labelled 96-well plate. The optical density was measured at 540 nm.

 

Data are presented in the form of percentage viability (MTT reduction in the test material treated tissues relative to the negative control tissues).

 

Results: The relative mean viability of the test material treated tissues was 102.2% after a 15 minute exposure.

 

Quality criteria: The quality criteria required for acceptance of results in the test were satisfied.

 

Conclusion: The test material was considered to be Non-Irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

May 17-27 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study (OECD 405, EU B.5)

Justification for data waiving:

other:

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Hillcrest, Belton, Loughborough, UK
- Age at study initiation: 12-20 weeks
- Weight at study initiation: 2.40 or2.42 kg
- Housing: individually housed in suspended cages
- Diet (e.g. ad libitum): ad libitum to 2030 Teklad global rabbit diet
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23 degrees C
- Humidity (%): 30-70%
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12 / 12 dark-light

IN-LIFE DATES: not reported

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 ml

VEHICLE: Not applicable

Duration of treatment / exposure:

Test material was applied to eye, and the eyelids were held together for about 1 second and then released. Eyes were not rinsed after exposure.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

2

Details on study design:

REMOVAL OF TEST SUBSTANCE: Not applicable

SCORING SYSTEM: initial pain reaction from 6-point scale, Draize scale for scoring ocular irritation was used during the observation period. After scores were assigned, a modified version of Kay and Calandra (1962) was used to classify the ocular irritancy potential.

TOOL USED TO ASSESS SCORE: ophthalmoscope

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 72 hours

Score:

0

Max. score:

12

Reversibility:

not specified

Remarks on result:

other: Both test animals had scores of 12 at 1 hour, which had diminished to 6 at 24 hours. Both had a score of 0 at 48 and 72 hours.

Irritant / corrosive response data:

See Table 1.

Other effects:

Body weight changes after 3 days: +0.05 kg

Table 1. Individual Scores and Individual Total Scores for Ocular Irritation Rabbit number and sex 69220 Male 69238 Male Time after treatment 1 hour 24 hours 48 hours 72 hours 1 hour 24 hours 48 hours 72 hours Cornea  E = Degree of opacity 0 0 0 0 0 0 0 0  F = Area of Cornea involved 0 0 0 0 0 0 0 0  Score (E x F) x 5 0 0 0 0 0 0 0 0 Iris  D 0 0 0 0 0 0 0 0  Score (D x 5) 0 0 0 0 0 0 0 0 Conjunctivae  A = Redness 2 1 0 0 2 1 0 0  B = Chemosis 2 1 0 0 2 1 0 0  C = Discharge 2 1 0 0 2 1 0 0  Score (A + B + C) x 2 12 6 0 0 12 6 0 0 Total Score 12 6 0 0 12 6 0 0

 

Interpretation of results:

slightly irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: modified Kay and Calandra 1962

Conclusions:

The test substance was determined to be a minimal irritant (Class 3 on a 1 - 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. .

Executive summary:

Study report summary:

Introduction. The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method was designed to meet the requirements of the following:

-      OECD guidelines for the testing of chemicals No. 405 “Acute Eye Irritation/Corrosion” (adopted 24 April 2002)

-      Method B.5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008

 

Result. A single application of the test material to the non-irrigated eye of two rabbits produced moderate conjuctival irritation. Both treated eyes appeared normal at the 48 -Hour observation.

 

Conclusion. The test material produced a maximum group mean score of 12.0 and was classified as a minimal irritant (Class 3 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.

 

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Some results for the whole family of fat liquors are reported. The whole family can be also compared to the family of Linear Alcan Sulphate/sulphonated and the fatty soap in general. In all those molcules, eye irritation is generally dependent to the molecular weight of the substance. being fatty acids derivatives the smaller molecular weight in the whole family, eye irritation can be assumedSkin and eye irritation data are available for the sodium salt of sulfated castor oil. These 1984 reasonably well documented studies conclude the test substance is not irritating to the skin but is irritating to the eye. Read across from similar substances (sulfited fat liquors EC 307-037-4 fish oil and EC 281-975-1 rape oil) indicate the substances are non-irritating and non-corrosive to the skin, and are slightly irritating to the eye based on OECD guideline studies.

Justification for selection of skin irritation / corrosion endpoint:
vegetal oil derivatives is the structural analogous more symilar to fatty acid vegetable and the first methabolite

Justification for selection of eye irritation endpoint:
vegetal oil derivatives is the structural analogous more symilar to fatty acid vegetable and the first methabolite

Effects on eye irritation: irritating

Justification for classification or non-classification

Based on existing studies the substance is classified as slightly irritant for the eyes, corresponding to a classification in category B and H 319 following Regulation 1272/2008 (CLP)