Registration Dossier

Administrative data

Description of key information

Skin irritation (in vivo): not irritating.

Eye irritation ( in vitro / ex vivo): not irritating.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin corrosion: in vitro / ex vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
(only basic information given; occlusive type of coverage, only 48 h observation period)
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
GLP compliance:
no
Species:
rabbit
Strain:
other: albino
Type of coverage:
occlusive
Preparation of test site:
clipped
Remarks:
abraded skin was additionally tested, but not used for evaluation
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
0.5 g
Duration of treatment / exposure:
4 h
Observation period:
48 h
Reading time points: 4, 24 and 48 h
Number of animals:
6
Details on study design:
TEST SITE
- Type of wrap if used: The patches (1.5 inch x 1.5 inch 12 ply gauze squares) were secured to the area by thin bands of adhesive tape. The entire trunks of the animals were then wrapped in clear plastic trunk bands.

REMOVAL OF TEST SUBSTANCE
- Washing: Test sides were washed with appropraite solvent
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
4, 24 and 48 h

SCORING SYSTEM:
- Method of calculation: Draize scoring system
Irritation parameter:
erythema score
Basis:
mean
Remarks:
of 6 animals
Time point:
24/48 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
edema score
Basis:
mean
Remarks:
of 6 animals
Time point:
24/48 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritant / corrosive response data:
Very slight edema were observed after the exposure period at 4 h on abraded skin in all animals. These effects were fully reversible within 24 h.
Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008
Conclusions:
Under the conditions of this skin irritation study the test substance was not irritating.
Executive summary:

In a test for skin irritation, rabbits were exposed to test material (test site occluded), following detailed examinations, post-exposure, the test substance was demonstrated not to be irritating.

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

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Circa 1973
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Qualifier:
no guideline followed
Principles of method if other than guideline:
Draize eye irritation test. Original reference source for Draize test: Draize J, Woodard G, Calvery H. 1944. Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membranes. J Pharm Exp Ther 82:377–390
GLP compliance:
no
Species:
rabbit
Strain:
not specified
Remarks:
Albino
Details on test animals or tissues and environmental conditions:
Not specified.
Vehicle:
not specified
Controls:
yes
Amount / concentration applied:
0.1 ml of DHA-Na sprayed.
Duration of treatment / exposure:
Spray 48 seconds.
Observation period (in vivo):
7 days.
Details on study design:
Sodium Dehydroacetate (100 percent) was analysed in a modified Draize eye irritation test. A 0.1 ml dose was sprayed for 48 seconds from a distance of 6 inches into the conjunctival sac of one eye of each of 6 albino rabbits. Eyes were graded and scored according to Draize at 1, 2, 3,4, and 7 days or until negative. Sodium Dehydroacetate had average irritation scores of 5, 1, 2, and 0 (max - 110) on Days 1, 2, 3, and 4, respectively.
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
24 h
Score:
ca. 1 - ca. 5
Max. score:
5
Reversibility:
fully reversible within: before 4 days
Remarks on result:
probability of weak irritation
Interpretation of results:
study cannot be used for classification
Conclusions:
Minimally irritating according to the Draize classification, reversible before 4 days after application.
Executive summary:

A Draize tested carried out in single rabbits on sodium dehydroacetate was minimally irritating and reversible before 4 days after application. The results suggests that sodium dehydroacetate does not warrant classification.

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06 Oct 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Version / remarks:
adopted Jul 2013
Deviations:
no
Qualifier:
according to
Guideline:
EU method B.48 (Isolated chicken eye test method for identifying occular corrosives and severe irritants)
Deviations:
not specified
GLP compliance:
yes (incl. certificate)
Remarks:
National Institute of Pharmacy and Nutrition, Budapest, Hungary
Species:
chicken
Strain:
other: ROSS 308
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: Taravis KFT., Sárvár, Hungary
- Storage, temperature and transport conditions of ocular tissue: Transport of the chicken heads approximately within 2 h from collection under ambient temperature (19.3 - 20.4 °C). After collection, the heads were inspected for appropriate quality, wrapped with paper moistened with saline and placed in closable box
- Time interval prior to initiating testing: 45 - 60 min
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.03 g

Duration of treatment / exposure:
10 s
Duration of post- treatment incubation (in vitro):
30, 75, 120, 180 and 240 min
Number of animals or in vitro replicates:
Number of eyes for the negative control: 1
Number of eyes for the positive control: 3
Number of eyes for the test substance: 3
Details on study design:
SELECTION AND PREPARATION OF ISOLATED EYES
Selection:The surface of the cornea was treated with fluorescein solution (2% (w/v)) for a few seconds and examined with slit lamp microscope to ensure that the cornea was not damaged.


EQUILIBRATION AND BASELINE RECORDINGS
At the end of the acclimatization period, a zero reference measurement was recorded for cornea thickness and opacity to serve as a baseline for each individual eye. The cornea thickness of the eyes should not change by more than ± 5 - 7% within approximately 45 - 60 min before the start of application. Changes in thickness were not observed in the eyes. Following the equilibration period, the fluorescein retention was measured. If any eye was considered to be unsuitable following baseline assessment, it was discarded.

NEGATIVE CONTROL USED
Saline solution (9 g/L)

POSITIVE CONTROL USED
Imidazole (0.03 g)

APPLICATION DOSE AND EXPOSURE TIME
0.03 g for test item and positive control, 30 uL for negative control; exposure period: 10 s

OBSERVATION PERIOD
30, 75, 120, 180 and 240 min

REMOVAL OF TEST SUBSTANCE
- Volume and washing procedure after exposure period: The cornea was rinsed thoroughly with 20 mL saline solution at ambient temperature

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Corneal opacity is evaluated by using the area of the cornea that is most densely opacified for scoring. The mean corneal opacity value for all test eyes is calculated for all observation time points. Based on the highest mean score for corneal opacity, as observed at any time point, an overall category score is then given for the test substance.
- Damage to epithelium based on fluorescein retention: Fluorescein retention is evaluated at the 30 minute observation time point. The mean fluorescein retention value of all test eyes is then calculated for the 30-minute observation time point, and used for the overall category score given for the test substance.
- Swelling: measured with optical pachymeter on a slit-lamp microscope
- Macroscopic morphological damage to the surface: subjective according to the interpretation of the study director

SCORING SYSTEM AND DECISION CRITERIA:
- Mean corneal swelling (%)
Cornea swelling: For the calculation of Maximum Swelling, small negative numbers for swelling (0 to -5%) following application are counted as zero. Large negative numbers (> -12%) are probably due to erosion and indicate a severe effect (scored as class IV). Cases of values of -5% to -12% are evaluated on a case by case basis but in the absence of other findings do not indicate a severe effect.

CS at time t : [(CT at time t - CT at t=0)/CT at t=0] x 100
Mean CS at time t = [FECS (at time t) + SECS (at time t) + TECS (at time t)] / 3

CS : Cornea swelling
CT : Cornea thickness
FECS : First eye cornea swelling
SECS : Second eye cornea swelling
TECS : Third eye cornea swelling
Mean CS : The mean percentage of corneal swelling
At time t : Observation time at 30, 75, 120, 180 or 240 min after the post-treatment rinse
At t=0 : Reference value

- Mean maximum opacity score
Score 0: No opacity
Score 0.5: Very faint opacity
Score 1: Scattered or diffuse areas; details of the iris are clearly visible
Score 2: Easily discernible translucent area; details of the iris are slightly obscured
Score 3: Severe corneal opacity; no specific details of the iris are visible; size of the pupil is barely discernible
Score 4: Complete corneal opacity; iris invisible

ΔCO at time t : CO at time t – CO at t=0
Mean CO (at time t) : [FEΔCO (at time t) + SEΔCO (at time t) + TEΔCO(at time t)]/3

CO : Cornea opacity
ΔCO : Difference between cornea opacity and cornea opacity reference value
FEΔCO : Difference between first eye cornea opacity and first eye cornea opacity reference value
SEΔCO : Difference between second eye cornea opacity and second eye cornea opacity reference value
TEΔCO : Difference between third eye cornea opacity and third eye cornea opacity reference value
Mean CO : The mean corneal opacity value
At time t : Observation time at 30, 75, 120, 180 or 240 min after the post-treatment rinse
At t=0 : Reference value

- Mean fluorescein retention score at 30 minutes post-treatment
Scores for fluorescein retention:
Score 0: No fluorescein retention
Score 0.5: Very minor single cell staining
Score 1: Single cell staining scattered throughout the treated area of the cornea
Score 2: Focal or confluent dense single cell staining
Score 3: Confluent large areas of the cornea retaining fluorescein

ΔFR at time t : FR at time t – FR at t=0
Mean FR (at time t) : [FEΔFR (at time t) + SEΔFR (at time t) + TEΔFR(at time t)]/3

FR : Fluorescein retention
ΔFR : Difference between fuorescein retention and fuorescein retention reference value
FEΔFR : Difference between first eye fuorescein retention and first eye fuorescein retention reference value
SEΔFR : Difference between second eye fuorescein retention and second eye fuorescein retention reference value
TEΔFR : Difference between third eye fuorescein retention and third eye fuorescein retention reference value
Mean FR : The mean fuorescein retention value
At time t : Observation time at 30 min after the post-treatment rinse
At t=0 : Reference value
Irritation parameter:
percent corneal swelling
Run / experiment:
75 min
Value:
1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
2%
Positive controls validity:
valid
Remarks:
24%
Irritation parameter:
percent corneal swelling
Run / experiment:
240 min
Value:
2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
2%
Positive controls validity:
valid
Remarks:
29%
Irritation parameter:
cornea opacity score
Run / experiment:
mean maximum o 3 eyes
Value:
0.3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
0.0
Positive controls validity:
valid
Remarks:
4.0
Irritation parameter:
fluorescein retention score
Run / experiment:
mean of 3 eyes
Value:
0.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
0.0
Positive controls validity:
valid
Remarks:
3.0
Other effects / acceptance of results:
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes

Table 1 Results of ICE Assay

Na-Dehydroacetic acid (0.03 g/eye)
Relative Observation time (min) Corneal thickness (instrument units) Corneal opacity score Fluorescein retention score
-45-60 0 Δ 30 75 120 180 240 0 30 75 120 180 240 0 30
5 59 59 0% 60 60 61 61 61 0 0 0 0.5 0.5 0.5 0 0.5
6 60 60 0% 60 60 60 60 60 0 0 0 0.5 0.5 0.5 0 0
7 60 60 0% 61 61 61 61 61 0 0 0 0 0 0 0 0
  Mean Swelling 1% 1% 2% 2% 2% MOS 0.0 0.0 0.3 0.3 0.3 MFRS 0.2
Imidazole (0.03 g/eye)
Relative Observation time (min) Corneal thickness (instrument units) Corneal opacity score Fluorescein retention score
-45-60 0 Δ 30 75 120 180 240 0 30 75 120 180 240 0 30
2 62 62 0% 70 74 74 75 77 0 4 4 4 4 4 0 3
3 60 60 0% 72 75 75 77 79 0 4 4 4 4 4 0 3
4 60 60 0% 73 76 78 79 79 0 3 4 4 4 4 0 3
  Mean Swelling 18% 24% 25% 27% 29% MOS 3.7 4.0 4.0 4.0 4.0 MFRS 3.0
NaCl (9g/L saline solution: 30 uL/eye)
Relative Observation time (min) Corneal thickness (instrument units) Corneal opacity score Fluorescein retention score
-45-60 0 Δ 30 75 120 180 240 0 30 75 120 180 240 0 30
2 60 60 0% 61 61 61 61 61 0 0 0 0 0 0 0 0.5
  Swelling 2% 2% 2% 2% 2% MOS 0 0 0 0 0 FRS 0.0

 Δ = cornea thickness change between the start of the acclimatization period (t = -60 to -45 min) and the baseline (t = 0)

MOS =  Mean opacity score

OS = Opacity score

MFRS = Mean fluorescein retention score

FRS = Fluorescein retention score

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008
Conclusions:
Under the conditions of this study the test material was shown not to be an eye irritant.
Executive summary:

In an isolated chicken eye test method conducted to assess potential eye irritation the test material was shown not to be irritating to the eye.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Skin irritation (in vivo): not irritating.: GHS criteria not met

Eye irritation ( in vitro / ex vivo): not irritating. : GHS criteria not met