4,4'-Isopropylidenediphenol, ethoxylated, esters with fatty acids, coco

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018-05-24 to 2018-06-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 442B (Skin Sensitization: Local Lymph Node Assay: BrdU-ELISA)

Version / remarks:

2010-07-22

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA): BrdU-ELISA

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-Isopropylidenediphenol, ethoxylated, esters with fatty acids, coco

EC Number:

500-303-2

EC Name:

4,4'-Isopropylidenediphenol, ethoxylated, esters with fatty acids, coco

Cas Number:

115340-85-7

IUPAC Name:

4,4'-Isopropylidenediphenol, ethoxylated, esters with fatty acids, coco

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: R-127. Lot 170825
- Expiration date of the lot/batch: 2019-07-26

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Solubility and stability of the test substance in the solvent/vehicle: soluble

In vivo test system

Test animals

Species:

mouse

Strain:

Balb/c

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: InVivos Pte Ltd., Singabpore
- Females (if applicable) nulliparous and non-pregnant: yes
- Microbiological status of animals, when known: Murine pathogen free (MPF)
- Age at study initiation: 9 weeks
- Weight at study initiation: 18.9 - 21.5 g
- Housing: OptiMICE Caging Systems
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 1 week
- Indication of any skin lesions: none

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 26 °C
- Humidity (%): 30 - 70 %
- IN-LIFE DATES: From: 2018-05-17 To: 2018-06-06

Study design: in vivo (LLNA)

Vehicle:

other: sesame oil, Ph Eur standard

Concentration:

50 % in vehicle.
25 µl administered to each ear

No. of animals per dose:

5

Details on study design:

Dose interval:
Topical application of 25 µl of test substance/negative control/positive control on the dorsum of each ear once a day for 3 consecutive days.

Administration of BrdU (10 mg/ml):
Administration: Intraperitoneal injection (IP)
Dose level: 5 mg (0.5ml of 10 mg/ml solution) per mice
Dose Interval: Single Dose

ELISA using BrdU colorimetric kit:
Approximately 24 hours after BrdU injection, the animals were euthanized using carbon dioxide. The draining auricular lymph nodes from each mouse ear were dissected and processed separately in phosphate buffered saline (PBS) for each animal. The dissected lymph nodes were trimmed fascia and fat, weighed, immediately used for Lymph node cells (LNC) preparation.
LNC suspension was prepared using #70 nylon cell strainer following the Test facility SOP. Approximately 25 ml of LNC was prepared per animal.
ELISA was performed with stop solution based on instruction manual of supplier (Roche Applied Science, Cat No 11 647 229 001) and Test Facility SOP.

Positive control substance(s):

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

Statistics:

Results for each test and control group were expressed as mean Stimulation Index (SI). The SI was derived by dividing the mean BrdU labelling index of test or positive control group by the mean BrdU labelling index of negative control group. The SI for negative control group was defined as 1.

BrdU labelling index (LI) = (Mean ABS450-Mean ABSblank 450) – (Mean ABS690-Mean
ABSblank 690)

SI = LI test (positive control) / LI negative control

Results and discussion

Positive control results:

SI = 1.692 (> 1.6); valid

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Remarks:

mean value

Value:

0.959

Results 2

Parameter:

SI

Remarks:

1st value

Value:

0.487

Results 3

Parameter:

SI

Remarks:

2nd value

Value:

1.451

Results 4

Parameter:

SI

Remarks:

3rd value

Value:

0.415

Results 5

Parameter:

SI

Remarks:

4th value

Value:

1.906

Results 6

Parameter:

SI

Remarks:

5th value

Value:

0.536

Cellular proliferation data / Observations:

CLINICAL OBSERVATIONS:
No adverse effect was observed on all the animals, except that very slight erythema at the dorsal part of the ears were observed in all the 5 animals of positive control group throughout all 3 induction days.
BODY WEIGHTS
Normal development

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the above study, using 50% (v/v) of the test item – 4, 4’-Isopropylidenediphenol, ethoxylated, esters with fatty acids, coco as test substance and sesame oil, Ph Eur standard as vehicle and negative control, no skin sensitization was produced in mice.

Executive summary:

The potential to cause skin sensitisation was examined in a study according to OECD 442 B.

Based on the above study, using 50% (v/v) of the test item – 4, 4’-Isopropylidenediphenol, ethoxylated, esters with fatty acids, coco as test substance and sesame oil, Ph Eur standard as vehicle and negative control, no skin sensitization was produced in mice.