Registration Dossier

Administrative data

Description of key information

Skin irritation: OECD 404: not irritating
Eye irritation: OECD 437: serious damage to eyes
Respiratory irritation: OECD 403: corrosive to respiratory tract (for further details refer to acute toxicity section)

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:
13 Mar 2011 - 18 Mar 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
(adopted 24 Feb 2002)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Bayerisches Landesamt fuer Gesundheit und Lebensmittelsicherheit, Erlangen, Germany
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Age at study initiation: 31-32 weeks
- Weight at study initiation: > 2 kg
- Housing: in ABS-plastic rabbit cages, floor 4200 cm²
- Diet: autoclaved hay and Altromin 2123 maintenance diet for rabbits (lot 1347), rich in crude fibre, ad libitum
- Water: tap water (drinking water, municipal residue control, microbiological controls at regular intervals), ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18±3
- Humidity (%): 55±10
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
water
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.5 g
Duration of treatment / exposure:
4 h
Observation period:
72 h
Number of animals:
3 females
Details on study design:
TEST SITE
- Area of exposure: dorsal area of trunk, approx. 6 cm²
- Type of wrap if used: patch was held in place with non-irritating tape and fixed with semi-occlusive dressing

REMOVAL OF TEST SUBSTANCE
- Washing: with tap water
- Time after start of exposure: 4 h

SCORING SYSTEM: Draize Scoring System
Irritation parameter:
erythema score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: reversibility: not applicable
Irritation parameter:
edema score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: reversibility: not applicable
Other effects:
There were no clinical signs of toxicity or any local effects noted in any of the animals at any of the reading time points.
The body weight development of animals was as expected.
Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008
Conclusions:
CLP: not classified
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:
09 Jan 2014 - 15 Jan 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Bayerisches Landesamt fuer Gesundheit und Lebensmittelsicherheit, Erlangen, Germany
Species:
other: cattle
Strain:
other: animals freshly slaughtered
Details on test animals or tissues and environmental conditions:
TEST METHOD
The bovine corneal opacity and permeability (BCOP) test is an in-vitro test method used for identifying i) chemicals inducing serious eye damage and ii) chemicals not requiring classification for eye irritation or serious eye damage. The potential of a test substance to cause ocular corrosivity or severe irritancy is measured by its ability to induce opacity and increased permeability in an isolated bovine cornea. The opacity and permeability assessments of the cornea are combined to derive an in-vitro irritancy score (IVIS), which is used to classify the irritancy level of the test substance.

IDENTIFICATION OF THE SOURCE OF THE EYES, STORAGE AND TRANSPORT CONDITIONS
- Source: A. Moksel AG, Buchloe, Germany
- Time interval prior to initiating testing: Cornea preparation was initiated immediately after arrival of the eyes.
- Transport medium and temperature conditions: Hanks´ Balanced Salt Solution (HBSS) on ice with penicillin/streptomycin

PREPARATION OF THE EYES (BEFORE EXPOSURE)
- Eyes free of defects (scratches, neovascularisation): yes
- Dissection of the eyes and treatment: Corneas were dissected with a 2 to 3 mm rim of sclera. Isolated corneas were mounted in cornea holders.
- Type of cornea holder used: MC2, Clermont, France
- Description of the cornea holder: The cornea holders consist of an anterior and a posterior compartment, which are tightened with screws.
- Test medium used in the cornea holder: RPMI (without phenol red) containing 1% FBS and 2 mM L-glutamine (complete RPMI)
- Equilibration time: 1 h at 32±1 °C
- Quality check of the equilibrated corneas: corneas with an initial opacity above 7 in the opacitometer were discarded.

DETERMINATION OF THE INITIAL OPACITY
- Method: Corneal opacity was determined by the amount of light transmission through the cornea via an opacitometer.
- Specification of the device: MC2, Clermont, France
Vehicle:
physiological saline
Remarks:
0.9% NaCl
Controls:
other: number of eyes for the negative control: 3
Amount / concentration applied:
TEST MATERIAL
- Amount applied (volume or weight with unit): 750 µL
- Concentration: 20% in vehicle

VEHICLE
- Substance: physiol. saline
- Concentration: 0.9% NaCl solution
- Source: Diprom
- Lot/batch no.: 13001-4
- Expiry date: 12/2015

POSITIVE SUBSTANCE
- Substance: Imidazole
- Concentration: 20% imidazole solution in physiol. saline
- Amount applied in the test: 750 µL
- Source: Sigma,
- Lot/batch no.: SLBC7446V
- Expiry date: 11/2014
Duration of treatment / exposure:
4 hours±5 minutes incubation at 32±1 °C
Observation period (in vivo):
Not applicable.
Number of animals or in vitro replicates:
Number of eyes for the test item: 3
Details on study design:
TEST CONDITIONS
- Short description of the method used: 750 µL of the test item preparation or the control substance was introduced into the anterior chamber (closed-chamber method). Corneas were exposed for 4 h at 32±1 °C with the test substance or the controls.

POST-EXPOSURE TREATMENT
- Removal of the test substance: The test substance was removed and the epithelium washed at least three times.
- Medium for washing the corneas: EMEM containing phenol red
- Medium for final rinsing: complete RPMI (without phenol red)

DETERMINATION OF THE FINAL OPACITY
- Method: Corneal opacity was determined by the amount of light transmission through the cornea via an opacitometer.
- Time of determination: After refilling complete RPMI in the anterior chamber the final opacity was measured.

DETERMINATION OF THE CORNEAL PERMEABILITY:
- Method: Sodium fluorescein solution was added to the anterior chamber of the cornea holder while the posterior chamber was filled with fresh complete RPMI. The amount of sodium fluorescein that crosses into the posterior chamber was quantitatively measured via UV/VIS spectrophotometry at 490 nm recorded as optical density (OD490).
- Amount and concentration of the dye: 1 mL sodium fluorescein solution (5 mg/mL)
- Incubation time: 90 min at 32±1 °C
Irritation parameter:
in vitro irritation score
Run / experiment:
mean value of 3 corneae
Value:
280.15
Vehicle controls validity:
valid
Remarks:
1.30
Negative controls validity:
not applicable
Positive controls validity:
valid
Remarks:
172.12

Tab. 1: Opacity values

Parameter

Cornea No.

Initial opacity

Final opacity

Change of opacity value

Corrected opacity value

Negative control

1

3

4

1

 

2

3

2

-1

3

3

2

-1

Mean

3.00

2.67

-0.33

Positive control

1

4

137

133

133.3

2

4

151

147

147.33

3

4

153

149

149.33

Mean

4.00

147.00

143.00

143.33

Test substance

1

3

330

327

327.33

2

3

251

248

248.33

3

3

268

265

265.33

Mean

3.00

283.00

280.00

280.33

 

Tab. 2: Permeability values (optical density (OD) at 490 nm)

Parameter

Cornea No.

OD490

Corrected OD490 value

Negative control

1

0.104

 

2

0.120

3

0.1102

Mean

0.109

Positive control

1

2.048

1.939

2

2.036

1.927

3

1.999

1.890

Mean

2.028

1.919

Test substance

1

0.134

0.025

2

0.110

0.001

3

0.046

-0.063

Mean

0.097

-0.012

 

Tab. 3: In-Vitro Irritancy Score (IVIS) values

Parameter

Cornea No.

Change of opacity value

Corrected OD490 value

IVIS

Negative control

1

1.00

0.104

1.30

2

-1.00

0.120

3

-1.00

0.1102

Mean

-0.33

0.109

Positive control

1

133.33

1.939

172.12

2

147.33

1.927

3

149.33

1.890

Mean

143.33

1.919

Test substance

1

327.33

0.025

280.15

2

248.33

0.001

3

265.33

-0.063

Mean

280.33

-0.012

 

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
CLP: Cat. 1, H318
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Additional information

Skin irritation/corrosion

A reliable key skin irritation /corrosion study with Reaction mass of ammonium diaqua[bis(oxalate)]oxoniobate(1-) hydrate and ammonium hydrogen oxalate oxalic acid (1:1:1) dihydrate is available, conducted according to OECD test guideline 404 (skin irritation in vivo), and in compliance with GLP (CBMM Europe BV, Key, 2011, OECD 404). Three female New Zealand White rabbits were semiocclusively applied 0.5 g of the test item dissolved in water on the dorsal area of the trunk (approx. 6 cm²) for 4 h. At the end of the exposure duration, residues of the test item were removed with tap water. The animals were observed for skin reactions according to the Draize Scoring System 1, 24, 48 and 72 h following end of exposure. There were no local effects noted in any of the animals at any of the reading time points. The body weight development of the animals was as expected and none of the animals showed any clinical signs of toxicity throughout the study period. Thus, the test item is concluded to be not irritating to the rabbits’ skin under the conditions of this test.

 

A supporting skin irritation/corrosion study with Reaction mass of ammonium diaqua[bis(oxalate)]oxoniobate(1-) hydrate and ammonium hydrogen oxalate oxalic acid (1:1:1) dihydrate is available, conducted in principle according to OECD test guideline 404 (skin irritation in vivo), but not in compliance with GLP (CBMM Europe BV, Supporting, 2006, OECD 404). Three rabbits of unknown strain were applied 0.5 g of the unchanged test item for 4 h. No information is given on the type of coverage in the study report. 4 h after start of exposure, residues of the test item were removed. The animals showed no skin irritation when observed 24, 48, and 72 h after treatment. Body weight development of all test animals was as expected and there were no clinical signs of toxicity noted in any of the animals throughout the study period. Thus, the test item is concluded to be not irritating to the rabbits’ skin under the conditions of this test.

 

Eye irritation

Eye irritation of Reaction mass of ammonium diaqua[bis(oxalate)]oxoniobate(1-) hydrate and ammonium hydrogen oxalate oxalic acid (1:1:1) dihydrate was investigated in an in-vitro bovine corneal opacity and permeability test, conducted according to OECD 437, and in compliance with GLP (CBMM Europe BV, Key, 2014, OECD 437). The bovine corneal opacity and permeability (BCOP) test is an in-vitro test method used for identifying chemicals inducing serious eye damage and chemicals not requiring classification for eye irritation or serious eye damage. The potential of a test substance to cause ocular corrosivity or severe irritancy is measured by its ability to induce opacity and increased permeability in an isolated bovine cornea. The opacity and permeability assessments of the cornea are combined to derive an in-vitro irritancy score (IVIS), which is used to classify the irritancy level of the test substance. In the present study, mean in-vitro irritancy scores calculated were 1.30, 280.15 and 172.12 for the negative control, the test item, and the positive control, respectively. The in-vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid. The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background opacity and permeability values for bovine corneas treated with the respective negative control. Thus, the test item is concluded to be corrosive to the bovine eye in the in-vitro bovine corneal opacity and permeability test. Reaction mass of ammonium diaqua[bis(oxalate)]oxoniobate(1-) hydrate and ammonium hydrogen oxalate oxalic acid (1:1:1) dihydrate meet therefore the criteria to be classified for inducing serious damage to eyes (Eye dam. 1).

 

Respiratory irritation

A reliable key acute inhalation toxicity study with Reaction mass of ammonium diaqua[bis(oxalate)]oxoniobate(1-) hydrate and ammonium hydrogen oxalate oxalic acid (1:1:1) dihydrate is available, conducted according to OECD test guideline 403, and in compliance with GLP (CBMM Europe BV, Key, 2014, AIT). Deaths observed in this study are contributed to local corrosive effects to the respiratory tract, indicated by observations such as irregular breathing, rales, nasal discharge, ocular discharge, gasping, etc., as well as gross necropsy findings (red lungs), which are observations commonly seen in animals suffering from local irritating/corrosive effects to the respiratory tract. Together with the facts that Reaction mass of ammonium diaqua[bis(oxalate)]oxoniobate(1-) hydrate and ammonium hydrogen oxalate oxalic acid (1:1:1) dehydrate is corrosive to eyes (mucous membranes) and the pH in aqueous solution is 0.4, the substance is concluded to be irritating to the respiratory tract. For further details please refer to the acute toxicity section.

Justification for classification or non-classification

The available data are reliable and suitable for classification. Based on these data, Reaction mass of ammonium diaqua[bis(oxalate)]oxoniobate(1-) hydrate and ammonium hydrogen oxalate oxalic acid (1:1:1) dihydrate meets the criteria to be classified for irreversible effects on the eye (Cat 1, H318) according to EC/1272/2008. Furthermore, based on expert judgement a labelling as STOT SE Cat 3 (H335 - May cause respiratory irritation) is warranted (for further details please refer to the acute toxicity section).