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: Follows a recognised guideline and performed to GLP standards.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
mouse
Strain:
other: CBA/Ca
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK limited, Bicester, Oxon, UK
- Age at study initiation: 8-12 weeks old
- Weight at study initiation: 15-23 g
- Housing: Suspended solid-floor polypropylene cages furnished with softwood woodflakes.
- Diet (e.g. ad libitum): ad libitum (2014 Teklad global rodent diet)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 C
- Humidity (%): 30-70%
- Air changes (per hr): 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark.

Study design: in vivo (LLNA)

Vehicle:
other: 1% pluronic L92 in distilled water.
Concentration:
25%, 10%, or 5% in 1% pluronic L92 in distilled water.
No. of animals per dose:
4 animals per dose.
Details on study design:
Preliminary test: A mouse was treated by daily application of 25 microlitres of trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate at a concentration of 25% w/w in 1% pluronic L92 in distilled water, to the dorsal surface of each ear for 3 consecutive days (days 1,2,3). The mouse was observed twice daily on day 1, 2, and 3 and once daily on days 4, 5, and 6. Signs of toxicity or excessive local irritation noted during this period were recorded.

Main test: Groups of 4 mice were treated with trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate at concentrations of 25%, 10% or 5% w/w in 1% pluronic L92 in distilled water. The preliminary test suggested that trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate would not produce systemic toxicity or excessive local irritation at the highest suitable concentration. The mice were treated by daily application of 25 microlitres of the appropriate concentration of trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate to the dorsal surface of each ear for 3 consecutive days (days 1, 2, 3). Trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate was administered using an automatic micropipette and spread over the dorsal surface of the ear using the tip of the pipette.
Positive control substance(s):
other: 2,4-Dinitrobenzenesulfonic acid, sodium salt

Results and discussion

Positive control results:
The stimulation index for the positive control can be found in table 1. 2,4-Dinitrobenzenesulfonic acid, sodium salt was considered to be a sensitiser.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: See table 2 for stimulation indexes.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Remark
Remarks:
Clinical observations, bodyweight and mortality data for the preliminary screening test can be seen in table 2. The radioactive disintegrations per minute per lymph node and the stimulation index are given in table 3. A stimulation index of less than 3 was recorded for the three concentrations of the test material (25%, 10% and 5% w/w in 1% pluronic L92 in distilled water). Table 4 shows clinical observations and mortality data from the main test.

Any other information on results incl. tables

Table 1. Stimulation index results for 2,4 -Dinitrobenzenesulfonic acid, sodium salt

 Concentration (% w/w) in 1% pluronic L92 in distilled water Stimulation index  Result 
0.96  Negative 
4.29  Positive 
10  13.15  Positive 

Table 2 Preliminary test: clinical observations, bodyweight and mortality data.

Concentration (%w/w) in 1% pluronic L92 in distilled water Animal number Bodyweight (g) Day Day
Day 1 Day 6 1 2 3 4 5 6
Pre-dose Post-dose Pre-dose Post-dose Pre-dose Post-dose      
25 S-1 22 23 0 0 0 0Rt 0 0Rt 0 0 0

(0 = no signs of systemic toxicity; Rt = Off white residual test material on the ears)

Table 3 Main test: Disintegrations per minute, Disintegrations per minute/node and stimulation index

 Concentration (% w/w) in 1% pluronic L92 in distilled water dpm  dpm/node (a)  Stimulation index (b)  Result 
Vehicle  2688.88  336.11  na  na 
5001.70  625.21  1.86  Negative 
10  2947.37  368.42  1.10  Negative 
25  3678.64  459.83  1.37  Negative 

(a) - Disintegrations per minute/node obtained by dividing the disintegrations per minute value by 8 (total number of lymph nodes)

(b) - Stimulation Index of 3.0 or greater indicates a positive result.

na - not applicable.

Table 4. Main test: individual clinical observations and mortality data.

Concentration (%w/w) in 1% pluronic L92 in distilled water Animal number Day
1 2 3 4 5 6
Pre-dose Post-dose Pre-dose Post-dose Pre-dose Post-dose 0 0 0
Vehicle 1-1 0 0 0 0 0 0 0 0 0
1-2 0 0 0 0 0 0 0 0 0
1-3 0 0 0 0 0 0 0 0 0
1-4 0 0 0 0 0 0 0 0 0
5 2-1 0 0 0 0 0 0 0 0 0
2-2 0 0 0 0 0 0 0 0 0
2-3 0 0 0 0 0 0 0 0 0
2-4 0 0 0 0 0 0 0 0 0
10 3-1 0 0 0 0 0 0Rt 0 0 0
3-2 0 0 0 0 0 0Rt 0 0 0
3-3 0 0 0 0 0 0Rt 0 0 0
3-4 0 0 0 0 0 0Rt 0 0 0
25 4-1 0 0 0 0Rt 0 0Rt 0 0 0
4-2 0 0 0 0Rt 0 0Rt 0 0 0
4-3 0 0 0 0Rt 0 0Rt 0 0 0
4-4 0 0 0 0Rt 0 0Rt 0 0 0

(0 = No signs of systemic toxicity; Rt = Off white residual test material on the ears)

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate was considered to be a non-sensitiser under the conditions of this test.
Executive summary:

In a dermal sensitization study with trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate in 1% pluronic L92 in distilled water, 12 female CBA/Ca mice (8 -12 week old) were tested using the method of Local Lymph Node Assay (LLNA). 2,4-Dinitrobenzenesulfonic acid, sodium salt was used as the positive control material. No clinical signs were observed during this study.                           

 

In this study, trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate is not a dermal sensitiser.