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Administrative data

Description of key information

Diammonium sodium hexakis(nitrito-N)rhodate was non-sensitising in a mouse local lymph node assay (LLNA), conducted according to OECD Test Guideline 429 and to GLP (Henzell, 2013).

 

No respiratory tract sensitisation data are available.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 August - 18 September 2013
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Guideline study performed largely to GLP (except that the stability of the test item was determined in a separate analytical study, and no analysis was conducted to determine the homogeneity or concentration of the test item). Minor deviations (humidity outside 30-70% range) not expected to affect study validity.
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
yes
Remarks:
on occasions, relative humidity was outside the target range of 30-70%. This was not considered to affect study integrity
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
GLP compliance:
yes (incl. certificate)
Remarks:
Except that the stability of the test item was determined in a separate analytical study, and no analysis was conducted to determine the homogeneity or concentration of the test item.
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
other: CBA/Ca (CBA/CaOlaHsd)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories Ltd., Oxon, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 15-23 g
- Housing: individually housed in suspended solid floor polypropylene cages furnished with softwood woodflakes
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): target: 19-25 (no data on actual values, but presumably not outside this range)
- Humidity (%): target: 30-70 (no data on actual values, but occasional deviations outside this range were noted, and were not considered to affect study integrity)
- Air changes (per hr): ~15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: no data
Vehicle:
propylene glycol
Concentration:
Range-finding study: 25% (w/w)
Main study: 0, 5, 10 or 25%
No. of animals per dose:
Range-finding study: 1
Main study: 5
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: solubility in various common vehicles assessed to find the vehicle giving the highest concentration suitable for dosing
- Irritation: yes. Range-finding mouse observed twice daily on days 1, 2 and 3, and once daily on days 4, 5 and 6, with erythema scored from 0-4.
- Lymph node proliferation response: not performed

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: local lymph node assay
- Criteria used to consider a positive response: the proliferation response of lymph node cells was expressed as the number of radioactive disintegrations per minute per animal and as the ratio of 3HTdR incorporation into lymph node cells of test nodes relative to that recorded for the control nodes (Stimulation Index). The test item was regarded as a sensitiser if at least one concentration resulted in a threefold or greater increase in 3HTdR incorporation compared to control values. Any test item failing to produce a threefold or greater increase in 3HTdR incorporation was classified as a "non sensitiser."

TREATMENT PREPARATION AND ADMINISTRATION: the test item was freshly prepared as a suspension in propylene glycol. The test item was formulated within two hours of being applied to the test system. 25 µl was applied topically to the ears of mice on days 1, 2 and 3 in both the range-finding and the main study.
Positive control substance(s):
other: phenylacetaldehyde (>90% purity) at 2.5% (v/v) in propylene glycol
Statistics:
Data were processed to give group mean values for disintegrations per minute and standard deviations where appropriate. Individual and group mean disintegrations per minute values were assessed for dose response relationships by analysis of homogeneity of variance followed by one way analysis of variance (ANOVA). In the event of a significant result from the ANOVA, pairwise comparisons were performed between control and treated groups. For homogenous datasets, Dunnett’s Multiple Comparison test was used, and for non homogenous datasets, Dunnett’s T3 Multiple Comparison Method was used.
Parameter:
SI
Remarks on result:
other: See Table 1 in "any other information on results incl. tables".
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: See Table 1 in "any other information on results incl. tables".

In the range-finding study, there were no signs of systemic toxicity, visual skin irritation, or 25% increased ear thickness (considered a sign of local irritation).

Table 1      Individual Disintegrations per Minute and Stimulation Indices

Treatment Group

Animal Number

dpm/
Animala

Mean dpm/Animal
(Standard Deviation)

Stimulation Indexb

Result

Vehicle
propylene glycol

1-1

1030.91

2038.00
(±812.38)

N/A

N/A

1-2

2227.51

1-3

3264.38

1-4

1801.09

1-5

1866.09

Test Item
5% w/w in
propylene glycol

2-1

1495.69

2421.31
(±646.90)

1.19

Negative

2-2

1993.61

2-3

2898.45

2-4

2752.74

2-5

2966.04

Test Item
10% w/w in
propylene glycol

3-1

1719.93

1949.44
(±493.19)

0.96

Negative

3-2

1565.86

3-3

2793.73

3-4

1963.41

3-5

1704.28

Test Item
25% w/w in
propylene glycol

4-1

1923.37

2781.17
(±763.81)

1.36

Negative

4-2

3983.18

4-3

2742.12

4-4

2399.61

4-5

2857.56

Positive Control Item

2.5% v/v in
propylene glycol

5-1

25068.56

12617.71
(±8237.59)

6.19

Positive

5-2

16448.63

5-3

10121.97

5-4

5236.74

5-5

6212.65

 

The results of the statistical analysis of the data indicated there was no significant difference between the control group and the test groups.

 

dpm=    Disintegrations per minute

N/A =     Not applicable

a=  Total number of lymph nodes per animal is 2

b=  Stimulation Index of 3.0 or greater indicates a positive result

Interpretation of results:
GHS criteria not met
Conclusions:
Diammonium sodium hexakis(nitrito-N)rhodate was non-sensitising in a mouse LLNA, conducted according to OECD guideline 429.
Executive summary:

The skin sensitising potential of diammonium sodium hexakis(nitrito-N)rhodate was assessed in a murine local lymph node assay (LLNA) conducted in accordance with OECD Test Guideline 429 and to GLP. Following a preliminary test using a concentration of 25% w/w, female CBA/Ca mice (5/group) received 25 µL applications of the test material, in propylene glycol at 0, 5, 10 or 25%, to the skin of both ears for three consecutive days. Two additional groups of 5 females were respectively treated with propylene glycol alone and phenylacetaldehyde, a known sensitiser, at 2.5%.

 

Mice were monitored for mortality and overt signs of toxicity, as well as changes in body weight. Three days after the induction period (on day six), mice were given tail vein injections of radiolabelled thymidine, and were sacrificed after approximately five hours. Lymph nodes were excised and processed for radioactivity.

 

No signs of skin irritation or systemic toxicity were observed in the preliminary test. In the main experiment, SI values were 1.19, 0.96 and 1.36 for animals given respective concentrations of 5%, 10% and 25%. An SI value of over 3 is judged to be indicative of a substance being sensitising. Hence, under the conditions of this assay, diammonium sodium hexakis(nitrito-N)rhodate was considered non-sensitising.

 

Based on the results of this study, diammonium sodium hexakis(nitrito-N)rhodate does not warrant classification for skin sensitisation, according to EU CLP criteria (EC 1272/2008).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

No relevant human skin sensitisation data were identified. No in vitro skin sensitisation studies were identified, or are required, as a reliable in vivo study is already available.

 

The skin sensitising potential of diammonium sodium hexakis(nitrito-N)rhodate was assessed in a mouse LLNA conducted in accordance with OECD Test Guideline 429 and to GLP. Following a preliminary test using a concentration of 25% w/w, female CBA/Ca mice (5/group) received 25 µL applications of the test material, in propylene glycol at 0, 5, 10 or 25%, to the skin of both ears for three consecutive days. Two additional groups of 5 females were respectively treated with propylene glycol alone and phenylacetaldehyde, a known sensitiser, at 2.5%. Mice were monitored for mortality and overt signs of toxicity, as well as changes in body weight. Three days after the induction period (on day six), mice were given tail vein injections of radiolabelled thymidine, and were sacrificed after approximately five hours. Lymph nodes were excised and processed for radioactivity. No signs of skin irritation or systemic toxicity were observed in the preliminary test. In the main experiment, SI values were 1.19, 0.96 and 1.36 for animals given respective concentrations of 5%, 10% and 25%. An SI value of over 3 was judged to be indicative of a substance being sensitising. Hence, under the conditions of this assay, diammonium sodium hexakis(nitrito-N)rhodate was considered non-sensitising (Henzell, 2013).

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:

No respiratory tract sensitisation data are available. A new study was not conducted as no standard and validated test method is available and it is not a REACH Standard Information Requirement.

Justification for classification or non-classification

Based on the results of the available and reliable murine LLNA assay, diammonium sodium hexakis(nitrito-N)rhodate does not warrant classification for skin sensitisation, according to EU CLP criteria (EC 1272/2008).