Trisodium 6,6',6''-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate

Skin sensitisation

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

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate

IUPAC Name:

trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate

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)

Results

Results 1

Parameter:

SI

Remarks on result:

other: See table 2 for stimulation indexes.

Results 2

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
1	0.96	Negative
5	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
5	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.