Distilled acetalization product between glucose and C12, C14, C18, C20 ,C22 alcohol

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: OECD guideline, GLP study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (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:

LCE 10047

IUPAC Name:

LCE 10047

Details on test material:

Test Substance/Itern : LCE 10047
Chemical name : Not applicable
(IUPAC)
Manufactured date : 18.11.2009
Retest date : 17.11.2012
Purity (as per Certificate Analysis) Acid value : 0.4 mg KOH/g
Hydroxyl value: 247 mg KOH/g

Physical appearance : Pearls appearance
Storage condition : Ambient (+18 to +36°C)
Note: Date of receipt of test item at test facility: 17.06.20 10 Test item code by test facility: S015-03

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

Species : Mice
Strain : CBA/Ca
Source : Harlan, The Netherlands
No. of groups : 7 groups
Vehicle control (G1)
Positive control (G2)
Test item: Five concentrations (G3 to G7)
Age at treatment : 11-12 weeks
Body weight range : 19.29 to 24.80 grams
Identification : Identification of individual animals: by cage card and body markings using turmeric solution.
Acclimatization : After physical examination for good health and the suitability for the study, the animals were acclimatized for 6 days before start of the treatment. Animals were nuliiparous and nonpregnant.

Husbandry
Room Number: Room No. SC-3
Conditions
Mice were housed in an environment controlled room at 21 to 23°C and relative humidity of 65 to 67 per cent. The photoperiod was 12 hours light and 12 hours darkness, light hours being 06.00 to 18.00 hours approximately. Adequate fresh air suppiy of 14.6 air changes/hour was maintained in the experimental room.
The maximum and minimum temperature and relative humidity in the experimental room were recorded once daily. The relative humidity in the experimental room was calculated daily from dry and wet bulb temperature recordings.

Housing
Animals were housed individually in solid floor standard polysulfone cages (Size: Approximately L 360 x B 205 x H 140 mm), with stainless steel top grill having facilities providing pelletted food and drinking water in polycarbonate bottle with stainless steel sipper tubes. Steam sterilized clean paddy husk was used as bedding and changed along with the cage twice a week. Cages were placed on 5 tier racks.

Diet and Water
The experimentai animals were provided ad libitum standard pelletted food (Ssniff mice peliet food — maintenance manufactured by Ssniff Spezialditen GmbH., Ferdinand-Gabriel-Weg 16, D-59494 Sôest, Germany) and deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-une water filter-cum-purifier (manufactured by Eureka Forbes Ltd., Mumbai -400001, India).
The food and water provided to the animals was tested for contaminants.

Study design: in vivo (LLNA)

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

positive control: 30% v/v

test material: 0.5 / 1 / 2.5 / 5 / 10 % w/v

No. of animals per dose:

5

Details on study design:

Grouping
Animals were randomly distributed to different groups by body weight stratification method. The mice procured for the study were weighed and grouped into body weight ranges. These body weight stratified animals were distributed to au the study groups. Mice with extreme body weights were removed from the study.

Dose Preparation and Analysis
Concentrations tested for the irritancy screen were selected based upon miscibility/solubility in an appropriate LLNA vehicle. The toxicity data regarding the irritation potential was also taken into consideration.
The required quantity of the test item was melted in hot water bath (60 - 70 °C). The test item LCE 10047 was mixed with AOO to obtain concentrations of 0.5, 1, 2.5, 5 and 10% w/v, vortexed and kept in water bath until application. The test item solutions were preparcd daily just prior to dosing. Preparation of the dosing materials was documented in the study file. The concentration of the dosing solution was not verified analytically.

Irritation Screen
Prior to the LLNA main study, concentrations of 0.1, 0.5, 1, 2.5, 5 and 10% w/v of LCE 10047, suspended in AOO were evaluated for irritation potential as measured by erythema of the ears.
Both ears of six female mice (one mouse/concentration) were topically treated once daily for three consecutive days with one of the concentrations of the test item.
The test item was administered using an adjustable micropipette with disposable tips. All mice received 25 uL (12 and 13 uL to each ear) of one concentration of the test item, spread over the dorsal surface of each ear in a manner to prevent test item loss. Similarly the vehicle was applied to the ears of one animal. Both ears were observed prior to application of the test item, and erythema was valuated on Days 2, 3 and 6. All mice were weighed on Days 1 and 6. Erythema scores and body weight data following the test item applications were compared to the response ofthe animals treated with vehicle alone (irritation to mouse ears is only relevant in the context of the LLNA and should not be interpreted as an indicator for an irritation potential in humans).

Dermal Sensitization
The application of the test item (25 uL/ear) was made on the dorsum of both ears as described above. Five female mice/group received the vehicle (AOO), or the positive control substance (30% a-hexylcinnamaldehyde), or 0.5, 1, 2.5, 5 and 10% w/v of LCE 10047, once daily for three consecutive days. Ears were inspected prior to each application of the test item solution, and erythema was evaluated on days 2, 3 and 6. All mice were weighed on days 1 and 6.

Positive control substance(s):

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

Results and discussion

Positive control results:

The sensitivity of this LLNA test was demonstrated via the response from the positive control (30% HCA in AOO), which elicited a stimulation index (SI) of 5.22, in comparison with the vehicle-treated mice.

In vivo (LLNA)

Results

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Average per group vehicle group: 1871.8 positive control group: 9773.6 0.5% group: 1636.2 1%: 2157.4 2.5%: 1974.4 5%: 2144.6 10%: 2198.2

Any other information on results incl. tables

negative control group: 1

positive control group: 5.22

0.5% group: 0.87

1%: 1.15

2.5%: 1.06

5%: 1.15

10%: 1.17

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

A topical application with 0.5, 1, 2.5, 5 and 10% w/v LCE 100467elicited a stimulation index (SI) of 0.87, 1.15, 1.06, 1.15 and 1.17 respectively. The test item LCE 10047 did not elicit a stimulation index (SI) that met the 3X threshold, thus indicating a lack of dermal sensitization potential in the mouse
LLNA.

Executive summary:

This Local Lymph Node Assay (LLNA) was conducted to evaluate the potential of LCE 10047 to cause contact sensitization by measuring lymphocyte proliferative response from auricular lymph nodes following topical application of the test item to the female CBA/Ca mouse ear.

Screening Study : Three daily topical application at 0.1, 0.5, 1, 2.5, 5 and 10% w/v of LCE 10047 in acetone:olive ou [AOO] (4:1 mixture v/v) were given to one animal at each dose level. There were no clinical signs of toxicity, no effect on body weight and no reactions at site of application. Results from this study were used to determine the dosing concentration for the main study.

LLNA main study : Five female CBA/Ca mice/group received the vehicle (AOO) or 30% a-hexylcinnamaldehyde (HCA: positive control in AOO) or 0.1, 0.5, 1, 2.5, 5 and 10%w/v

of LCE 10047 in AOO on days 1 to 3. On day 6, uptake of 3H-methyl thymidine into the auricular lymph nodes draining the site of test item application was measured five hours post administration. Proper conduct of the LLNA was confirmed via a positive response using 30% a-hexylcinnamaldehyde, a contact sensitizer, which elicited proliferation with a Stimulation Index (SI) value of 5.22, in comparison to vehicle-treated mice.

The test item LCE 10047 did not elicit erythema at the tested concentrations and no significant effect on body weight gain.

The test item LCE 10047 at dose concentrations of

0.1, 0.5, 1, 2.5, 5 and 10%

w/v elicited proliferative response with SI of 0.87, 1.15, 1.06, 1.15 and 1.17, respectively in comparison with the vehicle-treated mice.

LCE 10047 did not demonstrate dermal sensitization potential in the mouse LLNA, as the lymph nodes draining the area of topical application did not elicit a proliferative response greater than the 3X threshold.