Fatty acids, C6-19-branched, calcium salts

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018-07-16 to 2018-09-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2015-07-28

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

signed 2018-04-26

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C6-19-branched, calcium salts

EC Number:

270-064-4

EC Name:

Fatty acids, C6-19-branched, calcium salts

Cas Number:

68409-80-3

IUPAC Name:

Fatty acids, C6-19-branched, calcium salts

Test material form:

other: highly viscous, sticky

Details on test material:

- State of aggregation: highly viscous, sticky, yellow-brown

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature in a sealed container

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, 97633 Sulzfeld, Germany
- Age at study initiation: approx 45, 48 and 46 weeks old
- Weight at study initiation: 4.2 – 4.3 kg
- Housing: individually housed in ABS-plastic or Noryl rabbit cages, floor 4200 cm2
- Diet (ad libitum): autoclaved hay and Altromin 2123 maintenance diet for rabbits, rich in crude fibre
- Water (ad libitum): tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 18 ± 3 °C
- Relative humidity: 55 ± 10 %
- Air changes: at least 10 x / hour
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g of the test item
In order to ensure good skin contact, it was moistened with aqua ad injectionem.

Duration of treatment / exposure:

animal #1: 3 minutes, 1 hour and 4 hours
animal #2: 4 hours
animal #3: 4 hours

Observation period:

animal #1: 14 days
animal #2: 7 days
animal #3: 14 days

Number of animals:

3 male rabbits

Details on study design:

TEST SITE
- Area of exposure/Type of wrap if used: approx. 24 hours before the test, the fur was removed from the dorsal area of the trunk by using an electric clipper. The test item was applied to a small area (approx. 6 cm²) of skin on one side of the dorsal area and covered with a gauze patch, which was held in place with a non-irritating tape. The untreated other side served as control. The test item was applied to the patch first, moistened with the smallest amount of aqua ad injectionem and then applied to the skin. The patch was fixed with a semi-occlusive dressing. The limits of the application site were marked with an ink marker.

INITIAL AND CONFIRMATORY TESTING
As there was no information about the skin irritation/ corrosion potential of the test item, a single animal test was employed. Up to three test patches were applied sequentially to the animal. The first patch was removed after three minutes. No serious skin reaction was observed, so a second patch was applied at a different site and removed after one hour. The observations at this stage indicated that exposure could humanely be allowed to extend to four hours, so a third patch was applied and removed after four hours, and the response was graded. No corrosive effect was observed after the last patch was removed, so the animal was further observed.

The results of the initial test did not indicate the test item to be corrosive or a severe irritant to the skin using the procedure described. In order to confirm the response, two additional animals were treated in the same manner.

REMOVAL OF TEST SUBSTANCE
At the end of the exposure period, the residual test item was removed with sterile water from animal no. 1. As test item residues still stuck to the skin an appropriate solvent, cottonseed oil, was subsequently used daily to remove residual test material from animal no.1 and immediately after patch removal from animals no. 2 and 3 until no residues could be observed. Cottonseed oil was chosen in order not to alter the existing response or the integrity of the epidermis. The application site was rinsed daily before observation with cottonseed oil as test item residues were still observed.

OBSERVATION TIME POINTS
- animal #1: immediatley and 1 hour, 48, 72 and 96 hours after patch removal
- animal #2: 1 hour, 24, 48 and 72 hours after patch removal
- animal #3: 1 hour, 24, 48 and 72 hours after patch removal

SCORING SYSTEM
according to the Draize scale

FURTHER OBSERVATIONS
- body weights: prior to the administration and at the end of the observation period
- local effects such as hyperplasia, scaling, discolouration, fissures and scabs
- systemic effects

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Remarks:

animal #1

Time point:

other: 48, 72 and 96 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: Due to delayed symptom onset, the mean score is calculated from the values 48, 72 and 96 hours after patch removal

Results 2

Irritation parameter:

edema score

Basis:

mean

Remarks:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 3

Irritation parameter:

erythema score

Basis:

mean

Remarks:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 4

Irritation parameter:

edema score

Basis:

mean

Remarks:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 5

Irritation parameter:

erythema score

Basis:

mean

Remarks:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 8 days

Results 6

Irritation parameter:

edema score

Basis:

mean

Remarks:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

ANIMAL #1:
After patch removal during sequential application (3 min, 1 hour) erythema grade 1 was observed in animal no. 1. Immediately after patch removal after 4-hour exposure, no signs of irritation were noted in animal no. 1. Erythema grade 1 was again observed 48 hours after patch removal. Irritation score rose to erythema grade 2 from 72 hours until 6 days after exposure and decreased again to erythema grade 1 until day 13. 10, 11 and 12 days after exposure also slight scaling was observed at the application site. On the last day of observation, no signs of irritation were observed in animal no. 1. The observed effects were reversible within the observation period. The mean erythema score of animal no. 1, following grading at 48, 72 and 96 hours after installation of the test material (due to delayed onset of effects), was calculated to be 1.67.
ANIMAL #2:
Animal no. 2 showed erythema grade 1 at 24 and 72 hours and 6 days after patch removal. Erythema was increased to grade 2 at 48 hours, 4 and 5 days after patch removal. Erythema grade 1 together with scales at the application site were noted 6 days after exposure. The effects disappeared within 7 days after application. The mean erythema score of animal no. 2, following grading at 24, 48 and 72 hours post-application, was calculated to be 1.33.
ANIMAL #3:
Animal no. 3 showed erythema grade 2 from 24 hours until 5 days after patch removal. Erythema was decreased to grade 1 at 6 and 7 days after exposure. Erythema was reversible within 8 days after patch removal. Scaling was noted from 4 days after patch removal onwards and was not reversible but persisted until end of the observation period. The mean erythema score of animal no. 3, following grading at 24, 48 and 72 hours post-application, was calculated to be 2.00.

Other effects:

- Neither mortalities nor significant clinical signs of toxicity were observed.
- There were no significant body weight changes during the observation period, the body weight development was within the expected range.
- Residual test item at the application site was noted during sequential application in animal no. 1 and up to 72 hours after patch removal in the other both animals. Immediately after 4-hour exposure in animal no. 1 residual test item stuck to the application site and some adjoining fur was turn off upon patch removal.

Applicant's summary and conclusion

Interpretation of results:

other: not irritating to skin

Conclusions:

According to Regulation (EC) No 1272/2008 and subsequent adaptations, the test item is not irritating to skin and does not require classification and labelling.
According to UN GHS, the test item causes mild skin irritation and requires classification and labelling (Category 3).