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Toxicological information

Skin sensitisation

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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

Constituent 1
Chemical structure
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-isoindolo[2,1-a]perimidin-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)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: s. b.
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.