Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well documented study according to international accepted guidelines and GLP compliant.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Test item 4-nitrophenyl-acetic acid
Batch No. 0046D93
Physical state solid, crystalline powder
Colour yellowish to yellow-brownish
Storage cool, dry place, protected from light

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
Species and strain: CBA/Ca Ola Hsd mice
Source: TOXI-COOP ZRT.
Hygienic level at arrival: SPF
Hygienic level during the study: Good conventional
Number of animals: 24 animals/main test (4 animals/treatment group)
Sex: Female, nulliparous, non-pregnant
Age of animals: Young adult mice; 10-11 weeks old (at start of the main test)
Body weight range at starting: 17.6-22.7 g
Acclimatization time: 7 days
Animal health: Only healthy animals were used

Housing during acclimatization period: Grouped caging in small groups
Housing during the test: Grouped caging (4 animals/cage)
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 – 70 %

Study design: in vivo (LLNA)

Vehicle:
dimethylformamide
Remarks:
AOO (acetone:olive oil, 4:1) for positive control groups
Concentration:
The test item was administered at three different concentrations (25; 10; 5 w/v%) according to the results of the dose range finding test.
No. of animals per dose:
4 animal/dose
Details on study design:
Selection of test item concentrations was based on the results of a formulation evaluation and also results of preliminary irritation/toxicity test to find an appropriate solvent and the maximum applicable concentration according to the relevant guidelines. Based on the preliminary test results the test item was examined in the main test at 25 %, 10 % and 5 % (w/v) concentrations as formulations in the selected vehicle of N,N-Dimethylformamide (DMF). Appropriate positive control (α-Hexylcinnamaldehyde [HCA] in 25 % (w/v) concentration), furthermore two negative control groups dosed with the vehicles of the test and positive control groups, respectively, were employed.
The positive control item (25 % (w/v) HCA in Acetone: Olive oil 4:1 (v/v) mixture, AOO) induced the appropriate stimulation over the control, thus confirming the validity of the assay.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:
The positive control group animals were treated with 25 % (w/v) HCA solution (dissolved in AOO) concurrent to the test item groups. No mortality, cutaneous reactions or signs of toxicity were observed in the positive control group.
Significant lymphoproliferative response (SI ≥ 3) was noted for HCA (SI = 12.8). The results of the positive control item demonstrated appropriate performance of the test in accordance with the relevant guidelines and confirmed validity of the assay.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: see Remark
Remarks:
The stimulation index (SI = the DPN of a treated (positive control or test item) group divided by the DPN of the respective negative control group) for each treatment group was also calculated. A stimulation index of 3 or greater is an indication of a positive result. When monotonic dose-response is seen, attempt is made to calculate EC3 (dose calculated to induce a stimulation index of 3) with the help of linear regression on dose-response relationship. All calculations were made by Microsoft Excel Software. (See Table1 for the values.)
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Remark
Remarks:
DPM (disintegration per minute) was measured for each treatment group. The measured DPM values were corrected with the background DPM value. The average of the two measured DPM values of 5 % (w/v) TCA solutions was used as the background DPM value. The results were expressed as DPN (DPM divided by the number of pooled lymph nodes). (See Table1 for the values.)

Any other information on results incl. tables

Test Group Name

Measured

DPM/group

Group*

DPM

DPN

(DPM/Node)

Stimulation

Index Values

Vehicle control for the positive control: AOO

7283

7258.0

907.3

1.0

Positive control:
25 % HCA
in AOO

93261

93236.0

11654.5

12.8

4-nitrophenyl-acetic acid
25 %
in DMF

6763

6738.0

842.3

1.3

4-nitrophenyl-acetic acid
10 %
in DMF

5070

5045.0

630.6

0.9

4-nitrophenyl-acetic acid
5 % in DMF

7473

7448.0

931.0

1.4

Vehicle control for the test item: DMF

5397

5372.0

671.5

1.0

 *Group DPM = measured DPMgroup- average DPMbackground

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Migrated information
Conclusions:
In conclusion, under the conditions of the present Local Lymph Node Assay, 4-nitrophenyl-acetic acid (CAS No. 104-03-0) tested at the maximum attainable concentration of 25 % (based on solubility) and at concentrations of 10 % and 5 % (w/v) as formulations in an appropriate vehicle (DMF) was shown to have no skin sensitization potential.