Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1978
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: No GLP, short documentation, 24 h exposure time, purity not specified.
Qualifier:
equivalent or similar to guideline
Guideline:
other: [application time: 24 h (OECD TG 404: 4 h); 6 animals have been used (OECD: 3); observation period 8 days (OECD: 14 d); occlusive testing (OECD: semiocclusive); additionally scarified skin was tested (OECD: no abraded skin test required)]
Principles of method if other than guideline:
Method: §1500.41; Federal Register Vol. 38, No. 187, pp 26019 dated 27.09.1973
GLP compliance:
no
Remarks:
GLP was not compulsory at the time the study was conducted
Species:
rabbit
Strain:
Vienna White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Gaukler, Offenbach, Germany
- Weight at study initiation: 3.1 kg (2.45 kg - 3.43 kg)
- Diet: Ssniff K, standard diet, Intermast, Soest, Germany, ad libitum
- Water: ad libitum
Type of coverage:
occlusive
Preparation of test site:
other: clipped skin as well as clipped and scarified skin on the same animal
Vehicle:
water
Controls:
not required
Amount / concentration applied:
0.5 g of a 50% aqueous solution
Duration of treatment / exposure:
24 hour(s)
Observation period:
24h, 3 and 8 days
Number of animals:
6
Details on study design:
TEST SITE
- Area of exposure: 2.5 X 2.5 cm
- Type of wrap if used: The cloth on which the test substance was applied was covered with a gummed linnen cloth, which was fixed by sticking plasters and wrapped by a gauze bandage.

REMOVAL OF THE TEST SUBSTANCE
- no whashing done

ADDITIONAL INFORMATION
- During exposre animals were fixed and fasted.

SCORING SYSTEM:
- Draize Scale
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 24 and 72 h
Score:
1.1
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
other: Skin irritation results on the intact skin
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 24 and 72 h
Score:
0
Max. score:
4
Remarks on result:
other: Skin irritation results on the intact skin
Other effects:
- Four of the six test animals suffered from parasites in the liver

Reversible reddening was observed at the intact skin which
disappeared after three days. Mild to severe reddening and
edema was observed at the scarified skin. These effects were
reversible after 1 week and scale formation was observed.

Observation scores:

Intact skin:

Reddening:
time        score animal 1/2/3/4/5/6
24 h                     2/2/2/3/2/2
3 days                   0/0/0/0/0/0
8 days                   0/0/0/0/0/0
Oedema observation:
24 h                     0/0/0/0/0/0
3 days                   0/0/0/0/0/0
8 days                   0/0/0/0/0/0

Scarified skin

Reddening:
24 h                      2/3/3/3/2/2
3 days                    2/1/2/1/1/1
8 days                    0/0/0/0/0/0 scale formation in every case


Oedema observation:
24 h                       2/2/2/2/2/2
3 days                     2/0/2/0/1/0
8 days                     0/0/0/0/0/0

Conclusions:
slightly irritating
Executive summary:

500 mg of a 50 % aqueous suspension of adipic acid (99.8 %) was tested on intact and scarified skin of six rabbits, respectively. The compound was applied to an area of 5 x 5 cm, covered and held in contact for 24 hours. Responses were scored immediately after dosing (24 hours), 3 and 8 days. Reversible reddening was observed at the intact skin (scored 2-3 on a scale up to a maximum of 4) which disappeared after three days. Mild to severe reddening and edema was observed at the scarified skin (scores 24 h: 2, 3 days: 0 - 2). These effects were reversible after 1 week (all scores 0) and scar formation was observed.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06/2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP comparable to OECD guideline
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Qualifier:
according to guideline
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Qualifier:
according to guideline
Guideline:
other: Background Review Document (BRD): current status of in vitro test methods for identifying ocular corrosives and severe irritants: The Bovine Corneal Opacity and Permeability (BCOP) Test Method, March 2006.
Qualifier:
according to guideline
Guideline:
other: - In Vitro Techniques in Toxicology Database (INVITTOX) protocol 127. Bovine Opacity and Permeability (BCOP) Assay, 2006.
Qualifier:
according to guideline
Guideline:
other: - Gautheron P., Dukic M., Alix D. and Sina J.F., Bovine corneal opacity and permeability test: An in vitro assay of ocular irritancy. Fundam Appl Toxicol 18:442-449, 1992.
Principles of method if other than guideline:
/
GLP compliance:
yes
Species:
other: Cow
Strain:
other: not specified
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source:Bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands

ENVIRONMENTAL CONDITIONS
- Temperature (°C): corneas were stored at 32 +/- 1°C
Vehicle:
physiological saline
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Concentration (if solution): 20% (w/w) in physiological saline

VEHICLE
- Amount(s) applied (volume or weight with unit): 750µL
Duration of treatment / exposure:
240 +/- 10 mintues
Observation period (in vivo):
Opacity was determined without further incubation
Number of animals or in vitro replicates:
9 corneas were tested
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): After the incubation the solutions and the test compound were removed and the epithelium was washed at least three times with MEM with phenol red (Eagle’s Minimum Essential Medium, Invitrogen Corporation).
- Time after start of exposure: 240 min

SCORING SYSTEM:
OPACITY MEASUREMENT
The opacitometer determined the difference in the light transmission between each control or treated cornea and an air filled chamber. The numerical opacity value (arbitrary unit) was displayed and recorded. The change in opacity for each individual cornea (including the negative control) was calculated by subtracting the initial opacity reading from the final post-treatment reading. The corrected opacity for each positive control or test substance treated cornea was calculated by subtracting the average change in opacity of the negative control corneas from the change in opacity of each positive control or test substance treated cornea.
The mean opacity value of each treatment group was calculated by averaging the corrected opacity values of the treated corneas for each treatment group.
APPLICATION OF SODIUM FLUORESCEIN
Following the final opacity measurement, permeability of the cornea to Na-fluorescein (Merck) was evaluated.

The medium of both compartments (anterior compartment first) was removed. The posterior compartment was refilled with fresh cMEM. The anterior compartment was filled with 1 ml of 5 mg
Na-fluorescein/ml cMEM solution. The holders were slightly rotated, with the corneas maintained in a horizontal position, to ensure uniform distribution of the sodium-fluorescein solution over the entire cornea. Corneas were incubated in a horizontal position for 90 ± 5 minutes at 32 ± 1°C.
PERMEABILITY DETERMINATION
After the incubation period, the medium in the posterior compartment of each holder was removed and placed into a sampling tube labelled according to holder number. 360 μl of the medium from each sampling tube was transferred to a 96-well plate. The optical density at 490 nm (OD490) of each sampling tube was measured in triplicate using a microplate reader (TECAN Infinite® M200 Pro Plate Reader. Any OD490 that was 1.500 or higher was diluted to bring the OD490 into the acceptable range (linearity up to OD490 of 1.500 was verified before the start of the experiment). OD490 values of less than 1.500 were used in the permeability calculation.

The mean OD490 for each treatment was calculated using cMEM corrected OD490 values. If a dilution was performed, the OD490 of each reading was corrected for the mean negative control OD490 before the dilution factor was applied to the readings.
ELECTRONIC DATA CAPTURE
Observations/measurements in the study were recorded electronically using the following programme: Magellan Tracker 7.0 (TECAN, Austria) for optical density measurement.

IN VITRO IRRITANCY SCORE
The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:

In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).

Additionally the opacity and permeability values were evaluated independently to determine whether the test substance induced irritation through only one of the two endpoints.

A test substance that induces an IVIS ≥ 55.1 is defined as a corrosive or severe irritant.


TOOL USED TO ASSESS SCORE: fluorescein
Irritation parameter:
cornea opacity score
Value:
> 55.1
Remarks on result:
no indication of irritation
Interpretation of results:
slightly irritating
Remarks:
Criteria used for interpretation of results: OECD GHS
Conclusions:
It is concluded that this test is valid and that POTASSIUM ADIPATE is not severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.
Executive summary:

Screening for the eye irritancy potential of POTASSIUM ADIPATE using the Bovine Corneal Opacity and Permeability test (BCOP test).

 

This report describes the ocular irritation properties of POTASSIUM ADIPATE on an isolated bovine cornea. The possible ocular irritancy of POTASSIUM ADIPATE was tested through topical application for approximately 240 minutes.

 

The study procedures described in this report were based on the most recent OECD and EC guideline.

 

Batch 150413AK DRIED of POTASSIUM ADIPATE was a white crystalline powder with lumps.The test substance was applied as a 20% (w/w) solution (750 µl) directly on top of the corneas.

 

The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical rangeindicating that the negative control did not induce irritancy on the corneas.The meanin vitroirritancy score of the positive control (20% (w/v) Imidazole) was 150 and within the historical positive controldata range.It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

 

The meanin vitroirritancy score was 6.3 after 240 minutes of treatment with POTASSIUM ADIPATE.

Since the meanin vitroirritancy score for POTASSIUM ADIPATE was below 55.1 after 240 minutes treatment POTASSIUM ADIPATE is considered to be not severe irritant or corrosive.

 

Finally, it is concluded that this test is valid and that POTASSIUM ADIPATE is not severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for selection of skin irritation / corrosion endpoint:

The key study was selected

Justification for selection of eye irritation endpoint:

POTASSIUM ADIPATE is not severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.

Effects on eye irritation: slightly irritating

Justification for classification or non-classification

Potassium adipate is classified as Category 2B according to the CLP (EC) No. 1272/2008.