Reaction product of 6H-dibenz[c,e][1,2]oxaphosphorin 6-oxide, itaconic acid and ethylene glycol

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2008-11-28

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

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:

Bis(2-hydroxyethyl) (6H-dibenz[c,e][1,2]oxaphosphorin-6-ylmethyl)succinate P-oxide

EC Number:

264-313-6

EC Name:

Bis(2-hydroxyethyl) (6H-dibenz[c,e][1,2]oxaphosphorin-6-ylmethyl)succinate P-oxide

Cas Number:

63562-34-5

Molecular formula:

C21H23O8P

IUPAC Name:

Butanedioic acid, 2-​[(6-​oxido-​6H-​dibenz[c,​e]​[1,​2]​oxaphosphorin-​6-​yl)​methyl]​-​, 1,​4-​bis(2-​hydroxyethyl) ester

Details on test material:

Test item: UKANOL FR 50/1, EG-FREI

Chemical name:10-Dihydro-9-oxa-10-[2,3-Di-(2-hydroxyethoxy)carbonyl propyl]-10-phosphaphenanthren-10-oxid

CAS No.:63562-34-5 9

Name for the report:UKANOL FR 50/1, EG-FREI

Activity:100%

Test concentration: 10%, 25% and 50%

Description of the test item:brownish / glassy (observation by the study direction: solid, brittle)

In vivo test system

Test animals

Species:

mouse

Strain:

other: Crl:NMRI

Sex:

female

Details on test animals and environmental conditions:

The study was performed in 24 female SPF albino mice of the strain Crl:NMRI from the Charles River Deutschland GmbH, D-97633 Sulzfeld. At the time of the study the animals were about 9-10 weeks old and had a body weight ranging from 31 g to 36 g. An acclimatization period of at least 5 days was allowed.
The study took place in animal room No. 9 provided with filtered air at a temperature of 21°C ± 3°C, relative humidity being at least 30 % and preferably not exceed 70 % and air changes 10 times/ hour. The room was illuminated to give a cycle of 12 hours light and 12 hours darkness. Light was on from 6 am to 6 pm.

The mice were kept in groups in transparent macrolone cages (type 150, floor area 810 cm2) with six animals in each cage. The cages were cleaned and the bedding changed at least twice a week.

Bedding was "Lignocel-Fasern" from Altromin, D-32791 Lage, Lippe. Regular analyses for relevant possible contaminants are performed. Certificates of analysis are retained.

A pelleted complete rodent diet "Altromin 1314" from Altromin GmbH, D-32791 Lage, Lippe, was available ad libitum. Analyses for major nutritive components and relevant possible contaminants are performed regularly on the diet and certificates are retained.

The animals had free access to bottles with domestic quality drinking water acidified with hydrochloric acid to pH 2.5 in order to prevent microbial growth. Analyses for possible contaminants are performed regularly. Certificates of analysis are retained.

Study design: in vivo (LLNA)

Vehicle:

dimethyl sulphoxide

Concentration:

10%, 25% and 50%

No. of animals per dose:

6

Details on study design:

On day 1 the animals were weighed, the thickness of ears was measured by using a micrometer (Oditest S0247) and an amount of 25 pi of the test item or vehicle each was applied topically on the dorsal side of the ears.

The procedure as an open application was repeated on day 2 and 3 with the different concentrations of the test item or with the vehicle.

On day 4 the mice were weighed again and the thickness of ears was measured. Subsequently the animals were killed by inhalation of CO2 and a dissection with removal of the auricular lymph nodes was carried out. Furthermore circular tissue pieces with a diameter of 7 mm were punched from the ears of the animals in order to determine their weights. The lymph node pairs were also weighed and LN cell suspensions were prepared by mechanical tissue disruption.

The cell counts per millilitre of these suspensions were determined manually by trypan blue exclusion using a NEUBAUER-chamber and the LN cell counts were calculated from this.

Positive control substance(s):

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

Results and discussion

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

1.36

Test group / Remarks:

50 % test item

Results 2

Key result

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: LN cell count: Negative control: 12,326,667 50 % test item: 16,792,000 25 % test item: 14,360,000 10 % test item: 12,426,667

Results 3

Key result

Parameter:

SI

Value:

1.16

Test group / Remarks:

25% test item

Results 4

Key result

Parameter:

SI

Value:

1.01

Test group / Remarks:

10% test item

Results 5

Key result

Parameter:

SI

Value:

1

Test group / Remarks:

negative control

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the results of the study described neither a specific (sensitizing) nor a non-specific (irritant) stimulation potential shall be attributed to the test item UKANOL FR 50/1, EG-FREI in the tested concentrations of 10 %, 25 % and 50 %.

Executive summary:

The test item UKANOL FR 50/1, EG-FREI was investigated for the existence of a specific (sensitizing) or non-specific (irritant) stimulation potential by using the modified Local Lymph Node Assay (IMDS). Twenty-four albino mice of the strain Crl:NMRI in groups consisting of six animals each were treated with three concentrations of the test item UKANOL FR 50/1, EG-FREI [10 %, 25 % and 50 %] or only with the vehicle DMSO on three consecutive days. An amount of 25 µl of the test substances was applied on the dorsal side of each ear. The determination of ear thickness, ear weight, lymph node weight and lymph node cell count compared to the negative control (vehicle group) should provide the information whether the test item has a specific (sensitizing) or nonspecific (irritant) stimulation potential. In the test group treated with the 50 % test item 1 out of 6 mice was excluded from the study evaluation. It was not possible to remove the lymph nodes of animal No. 5 completely. Therefore, its data were not considered for evaluation and calculation of indices. In any case at least four animals per group were available. Within the study on day 4 there was no appreciable increase in ear thickness measured at the test groups animals compared with the negative control. The determination of ear weights did not show an appreciable increase at the test groups animals compared with the negative control either. The lymph nodes of the test group animals with test concentrations of 25 % and 50 % had a higher weight compared with the negative control. The cell counting of lymph node cells showed an increase of proliferation at the animals of the 25 % and 50 % test groups compared with the negative control. But on no account the treatment with the test item did lead to the exceeding of the positive threshold values. Since the positive threshold values for the increase of ear thickness and increase of lymph node cell (LN cells) were not exceeded through the treatment with the 10 %, 25 % and 50 % test item the calculation of the differentiation indices was not necessary. Based on the results of the study described in this final report neither a specific (sensitizing) nor a non-specific (irritant) stimulation potential shall be attributed to the test item UKANOL FR 50/1, EG-FREI in the tested concentrations of 10 %, 25 % and 50 %.