12H-phthaloperin-12-one

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

12H-phthaloperin-12-one

EC Number:

230-049-5

EC Name:

12H-phthaloperin-12-one

Cas Number:

6925-69-5

Molecular formula:

C18H10N2O

IUPAC Name:

12H-phthaloperin-12-one

Test material form:

solid

Details on test material:

orange colour
Insoluble in water
Storage at room temperature, protected from moisture
Expiration date: July 21, 2007

Specific details on test material used for the study:

orange solid

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands; B.V. Postbus 6174; NL 5960 AD Horst/ The Netherlands
- Age at study initiation: 8 -12 weeks
- Weight at study initiation: 14 - 19,3 g
- Housing: groups of four in Makrolon type-3 cagas
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 /12

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

0; 2.5 %, 5% and 10 %

No. of animals per dose:

4

Details on study design:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period: 3 days
- Test groups: 3
- Control group: 1
- Site: back of ear lobe
- Frequency of applications: 1/ day
- Concentrations: s.a.

Positive control substance(s):

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

Results and discussion

Positive control results:

ALPHA·HEXYLCINNAM-ALDEHYDE showed an allergenic potency when tested at concentration of 25 % (w/v).
EC3 is the estimated concentration for a STIMULATION INDEX of 3. In this study EC3 of 11 .3% (w/v) was theoretically calculated with STIMULATION INDICES of 2.6 and 7.1 at test item concentrations of 10 % and 25% (w/v).

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: s. b.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: s.b.

Any other information on results incl. tables

Test Item Concentration % (w/v)		Measurement dpm	Calculation			Result
			dpm - BG	Number of lymph nodes	dpm / lymph node	SI
	BG I	1	-	-	-	-
	BG II	1	-	-	-	-
0	CG 1	8186	8185	8	1023	-
2,5	TG 2	10841	10840	8	1355	1,3
5	TG 3	6286	6285	8	786	0,8
10	TG 4	5066	5065	8	633	0,6

 

BG = Background (1 ml 5% trichloro-acetic acid) in duplicate

CG = Control Group

TG = Test Group

SI = Stimulation Index

a) = The mean value was taken from the figures BG I and BG II

b) = Since the lymph nodes of the animals of a dose group were pooled, dpm/node was determined by dividing the measured value by the number of lymph nodes pooled

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was found to be a no sensitiser when apllied up to highest achievable concentration of 10 % (w/v) in acetone:olive oil, 4: I (v/v).
The test item caused slight local lrritation at ocncentrations of 2.5 %, 5% and 10 % (w/v).

Executive summary:

In order to study a possible allergenic potential of the test item three groups of four female mice each were treated with the test item at concentrations of 2.5 %, 5% and 10% (w/v) in acetone:olive oil, 4:1 (v/v) by topical application to the dorsum of each ear lobe (left and right) on three consequtive days. 10 % was the highest technically achievable concentration in the vehicle. A control group of four mice was treated with the vehicle (acetone:olive oil, 4:1 (v/v)) only. Five days after the first topical application the mice were injected intravenously into a tail vein with radio-labelled thymidine (³H·methyl thymidine).

Apploximately five hours after intravenouse injection, the mice were sacrificed, the draining lymph nodes excised and pooled per group.

Single cell suspensions of lymph nodes cells were prepared from pooled lymph nodes which were washed subsequently and incubated with trichloro acetic acid overnight. The proliferation of the pooled lymph node cells was determined by the incorporation of ³H-methyl-thymidine in a ß-scintillation counter.

No test item-relatad clinical signs were observed in any animals of the control group. Two hours after the first topical application, slight swelling and erythema were observed at both dosing sites in all mice of group 2 (2.5 %), group 3 (5 %) and group 4 (10 %), persisting for four to five days.

AlI treated animals survived the scheduled study period.

A test item is regarded as a Sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of ³HTdR at least 3-fold or greater than that recorded in control mice, as indicated by the stimultation index (SI).

 

	Test item concentration % (w/v)	SI
Group 2	2,5	1,3
Group 3	5	0,8
Group 4	10	0,6
No positive dose-response relation was observed. Calculation of the EC3 value was not done because no test concentration produced a stimulation index of 3 or higher.