6-methyl-1,2,3-oxathiazin-4(3H)-one 2,2-dioxide, potassium salt

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Remarks:

Local Lymph Node Assay in the Mouse with Acesulfame Potassium

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-Jul-12 through 2012-Sep-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

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:

6-methyl-1,2,3-oxathiazine-4(3-H)-one-2,2-dioxide potassium salt

IUPAC Name:

6-methyl-1,2,3-oxathiazine-4(3-H)-one-2,2-dioxide potassium salt

Constituent 2

Reference substance name:

6-methyl-1,2,3-oxathiazin-4(3H)-one 2,2-dioxide, potassium salt

EC Number:

259-715-3

EC Name:

6-methyl-1,2,3-oxathiazin-4(3H)-one 2,2-dioxide, potassium salt

Cas Number:

55589-62-3

Molecular formula:

C4H5NO4S.K

IUPAC Name:

potassium 6-methyl-2,2,4-trioxo-3,4-dihydro-1,2λ⁶,3-oxathiazin-3-ide

Constituent 3

Reference substance name:

Acesulfame potassium

IUPAC Name:

Acesulfame potassium

Test material form:

solid: crystalline

Details on test material:

- Name of test material (as cited in study report): Acesulfame potassium
- Physical state: White, crystalline powder
- Analytical purity:100%
- Lot/batch No.: 0000522972
- Expiration date of the lot/batch: 08 October 2012
- Stability under test conditions: stable
- Storage condition of test material: Controlled Room temperature (15-25 °C, below 70 RH%), protected from humidity

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: ELEVAGE JANVIER; Route des Chènes Secs B.P. 4105; 53940 LE GENEST-ST-ISLE, France
- Age at study initiation: 10 weeks
- Weight at study initiation: 20.7 - 22.8 g
- Housing: group caging
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 20 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70
- Air changes (per hr): 15 - 20
- Photoperiod (hrs dark / hrs light) 12 h/12h

IN-LIFE DATES:
From: 19-Sep-2012
To: 25-Sep-2012

Study design: in vivo (LLNA)

Vehicle:

dimethyl sulphoxide

Concentration:

5%, 10%, 25% (w/v)

No. of animals per dose:

4

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: soluble at 25% (w/v)
- Irritation: none at 25% (w/v)


MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method:
- Criteria used to consider a positive response: the DPN value of the negative (vehicle) control group falls within the range of historical laboratory control data, the positive control substance produces a significant lympho-proliferative response increases (SI>3), each treated and control group includes at least 4 animals, the test item did not cause serious systemic or local toxicity.

TREATMENT PREPARATION AND ADMINISTRATION:
Groups of four female CBA/J Rj mice were treated with: 25, 10 and 5% (w/v), respectively Acesulfame Potassium in DMSO. The negative control group received DMSO and the positive control group received 25 % -Hexylcinnamaldehyde (HCA) in DMSO.
The solutions of the test item were applied on the dorsal surface of ears of experimental animals (25 µl/ear) for three consecutive days (Days 1, 2 and 3). There was no treatment on Days 4, 5 and 6. On Day 6, the cell proliferation in the local lymph nodes was measured by incorporation of tritiated methyl thymidine (3HTdR) and the values obtained were used to calculate stimulation indices (SI).

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

Not applicable

Results and discussion

Positive control results:

α-Hexylcinnamaldehyde (25% (w/v) dissolved in DMSO) was used as a positive control to demonstrate the appropriate performance of the assay [1]. A significant lymphoproliferative response (stimulation index value of 15.7) was noted for the positive control chemical and this result confirmed the validity of the assay.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: Stimulation index values of the test item were 1.3, 1.4, and 1.5 at treatment concentrations of 25, 10 and 5% (w/v), respectively.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: DPM/group were were 1061, 1195 and 1275 at treatment concentrations of 25, 10 and 5% (w/v), respectively.

Results 3

Parameter:

SI

Value:

1.3

Results 4

Parameter:

SI

Value:

1.4

Results 5

Parameter:

SI

Value:

1.5

Any other information on results incl. tables

Body Weights

 

Animal	Test Group	Initial Body	Terminal Body	Change#
Number	Name	Weight (g)	Weight (g)*	(%)
424		22.8	22.7	-0.4
456	Negative control	21.8	23.8	9.2
478	DMSO	21.7	21.7	0.0
481		21.0	22.5	7.1
	Mean	21.8	22.7	3.9
427		22.3	23.4	4.9
451	Acesulfame Potassium	21.8	21.7	-0.5
460	25% (w/v) in DMSO	21.2	23.0	8.5
462		21.4	21.0	-1.9
	Mean	21.7	22.3	2.8
449		22.7	22.7	0.0
474	Acesulfame Potassium	20.7	20.5	-1.0
468	10% (w/v) in DMSO	21.4	23.1	7.9
452		21.3	20.5	-3.8
	Mean	21.5	21.7	0.8
466		22.0	22.7	3.2
454	Acesulfame Potassium	22.4	22.9	2.2
470	5% (w/v) in DMSO	21.0	21.9	4.3
480		20.9	21.8	4.3
	Mean	21.6	22.3	3.5
477		22.4	21.5	-4.0
484	Positive control	21.5	21.7	0.9
455	25 % HCA in DMSO	21.4	23.9	11.7
469		20.8	22.4	7.7
	Mean	21.5	22.4	4.1
*: Terminal body weights were measured on Day 6. #: (Terminal Body Weight – Initial Body Weight) / Initial Body Weight x 100

 

Desintegration per minute (DPM), desintegration per node (DPN) and Stimulation Index Values for all Groups:

Test Group	Measured	Group	No. of	DPN	Stimulation
Name	DPM/group	DPM	Nodes		Index Values
Background
(5 (w/v) % TCA )	37		-
Negative control
DMSO	850	813	8	101.6	1.0
Acesulfame Potassium
25% (w/v) in DMSO	1061	1024	8	128.0	1.3
Acesulfame Potassium
10% (w/v) in DMSO	1195	1158	8	144.8	1.4
Acesulfame Potassium
5% (w/v) in DMSO	1275	1238	8	154.8	1.5
Positive control
25 % HCA in DMSO	12789	12752	8	1594.0	15.7

 

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Acesulfame potassium tested in dimethyl sulphoxide as vehicle, was shown to have no sensitization potential in the Local Lymph Node Assay in female CBA/J Rj mice .

According to the UN Globally Harmonised System of Classification and Labelling of Chemicals, Acesulfame Potassium does not require classification as a skin sensitiser.

Executive summary:

The potential of Acesulfame potassium to induce skin sensitization was investigated in groups of four female CBA/J Rj mice treated with: 25, 10 and 5% (w/v) in DMSO. The negative control group received DMSO and the positive control group received 25 %a-Hexylcinnamaldehyde (HCA) in DMSO.

The solutions of the test item were applied on the dorsal surface of ears of experimental animals (25 µl/ear) for three consecutive days (Days 1, 2 and 3). There was no treatment on Days 4, 5 and 6. On Day 6, the cell proliferation in the local lymph nodes was measured by incorporation of tritiated methyl thymidine (³HTdR) and the values obtained were used to calculate stimulation indices (SI).

No mortality or sign of systemic toxicity or local irritation were observed during the study. No treatment body weight loss was observed during the study.

Stimulation index values of the test item were 1.3, 1.4, and 1.5 at treatment concentrations of 25, 10 and 5% (w/v), respectively.

The positive control demonstrated the appropriate performance of the assay as a significant lymphoproliferative response (stimulation index value of 15.7) was noted.

Conclusion

Acesulfame Potassium tested for its sensitizing potential according to OECD 429 under GLP conditions was shown to have no sensitization potential in the murine Local Lymph Node Assay.

The study result triggers the following classification/labelling:

EU Directive 1999/45/EC (as amended):        none

Regulation (EC) No 1272/2008 (CLP):           none

GHS (rev. 4) 2011:                                      unclassified