Quinoline

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: GLP study performed according to the OECD guideline.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

during the acclimation phase the relative humidity in the animal room was between 27 and 65% instead of 45-65%. This has no impact on the validity of the study.

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:

Quinoline

EC Number:

202-051-6

EC Name:

Quinoline

Cas Number:

91-22-5

Molecular formula:

C9H7N

IUPAC Name:

quinoline

In vivo test system

Test animals

Species:

mouse

Strain:

other: CBA/CaOlaHsd

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan laboratories BV
- Age at study initiation: 8-12 weeks
- Weight at study initiation: between 17.4 and 24.3 g
- Housing: single caging, in Makrolon type II cages, with wire mesh top. Bedding: granulated soft wood.
- Diet: pelleted standard diet, ad libitum
- Water: tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C +/- 2°C
- Humidity (%): 27-65%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 16-MAR-2010 to 14-APR-2010

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

0.5; 1; 2.5; 5 and 10%

No. of animals per dose:

4

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: pre-test on solubility showed that the highest item concentration was 100%
- Irritation: due to the acute toxicity of the substance the highest concentration tested in the pre-test was 25%. The 2 mice died, so a new pre-test was performed with 10%. Sign of local irritation were not observed, reduced spontaneous activity was observed within 1 hour after treatment, and not seen afterwards.
- Lymph node proliferation response: not determined

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response:
A test item is regarded as a sensitiser in the LLNA if the following criteria are fulfilled:
- First, that 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 stimulation index.
- Second, that the data are compatible with a conventional dose response, although allowance must be made (especially at high topical concentrations) for either local toxicity or immunological suppression


TREATMENT PREPARATION AND ADMINISTRATION:
The test item was placed into a volumetric flask on a tared balance and acetone:olive oil (4+1) was quantitatively added. The different test item concentrations were prepared serially.
The preparations were made freshly before each dosing occasion.
A correction factor of 1.02 for the purity was used

Each test group of mice was treated by topical (epidermal) application to the dorsal surface of each ear (left and right) with test item concentrations of 0.5, 1, 2.5, 5, and 10% (w/w) in acetone:olive oil (4+1). The application volume, 25 µl, was spread over the entire dorsal surface (  8 mm) of each ear once daily for three consecutive days. A further group of mice was treated with an equivalent volume of the vehicle (acetone:olive oil (4+1)) alone (control animals).

Positive control substance(s):

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

Statistics:

The mean values and standard deviations were calculated in the body weight tables. The ANOVA (Dunnett-test) was conducted on the ear thickness to assess whether the difference is statistically significant between test item groups and negative control (vehicle) group. However, both biological and statistical significance were considered together.

Results and discussion

Positive control results:

test performed in Nov 2009.
EC3 value is 12.9%

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: see table 1

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: see table 1

Any other information on results incl. tables

Table 1: calculation and results of individual data

Vehicle: acetone:olive oil (4+1)

Test item concentration % (w/w)	Group	Measurement DPM	Calculation			Result
			DPM-BG^a)	number of lymph nodes	DPM per lymph node^b^)	S.I.
---	BG I	30	---	---	---	---
---	BG II	62	---	---	---	---
---	1	5461	5415	8	676.9
0.5	2	5822	5776	8	722.0	1.07
1	3	6324	6278	8	784.8	1.16
2.5	4	6890	6844	8	855.5	1.26
5	5	9263	9217	8	1152.1	1.70
10	6	6063	6017	8	752.1	1.11

BG =  Background (1 ml 5% trichloroacetic acid) in duplicate

1    =  Control Group

2-6=  Test Groups

S.I. =  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

The EC3 value could not be calculated, since all S.I.´s are below 3.

The measured ear thickness of all animals treated was recorded prior to the 1^stapplication and prior to necropsy. A relevant increase in ear thickness was not observed in the animals treated with different test item concentrations in comparison to the control group.

Table 2: ear thickness

Animal No.	Ear thickness before the 1^st application (µm)				Ear thickness prior to treatment with^3HTdR (µm)				Ear thickness gain (µm)
	right	left	mean	Mean dose group±SD	right	left	mean	Mean dose group±SD	Thick-ness gain	Mean dose group
1	250	250	250.0	250.0±3.8	255	250	252.5	253.8±4.3	2.5	3.8±3.2
2	255	240	247.5		255	250	252.5		5.0
3	250	255	252.5		260	260	260.0		7.5
4	250	250	250.0		255	245	250.0		0.0
5	255	260	257.5	250.0±5.4	260	265	262.5	253.1±8.8	5.0	3.1±4.3
6	250	245	247.5		250	250	250.0		2.5
7	245	245	245.0		240	245	242.5		-2.5
8	250	250	250.0		255	260	257.5		7.5
9	250	250	250.0	245.0±3.5	260	255	257.5	253.8±4.8	7.5	8.8±4.3
10	245	245	245.0		250	255	252.5		7.5
11	240	245	242.5		250	245	247.5		5.0
12	245	240	242.5		260	255	257.5		15.0
13	250	255	252.5	252.5±2.0	255	250	252.5	254.4±3.8	0.0	1.9±4.3
14	255	255	255.0		250	255	252.5		-2.5
15	250	255	252.5		260	260	260.0		7.5
16	250	250	250.0		255	250	252.5		2.5
17	245	250	247.5	246.3±4.3	250	255	252.5	251.9±5.2	5.0	5.6±1.3
18	250	245	247.5		255	250	252.5		5.0
19	245	255	250.0		255	260	257.5		7.5
20	240	240	240.0		250	240	245.0		5.0
21	250	245	247.5	249.4±2.4	245	250	247.5	255.0±5.4	0.0	5.6±3.8
22	245	250	247.5		255	255	255.0		7.5
23	250	250	250.0		260	255	257.5		7.5
24	250	255	252.5		255	265	260.0		7.5

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

Quinoline is not a skin sensitizer.

Executive summary:

In the study, the test item Quinoline dissolved in acetone:olive oil (4+1) was assessed for its possible contact allergenic potential.

For this purpose a local lymph node assay was performed using test item concentrations of 0.5, 1, 2.5, 5, and 10%.

The animals did not show any clinical signs during the course of the study and no cases of mortality were observed. A statistically significant increase in ear thickness was not observed in the animals treated with different test item concentrations in comparison to the control group.

In this study Stimulation Indices (S.I.) of 1.07, 1.16, 1.26, 1.70, and 1.11 were determined with the test item at concentrations of 0.5, 1, 2.5, 5, and 10% in acetone:olive oil (4+1), respectively.

The test item Quinoline was not a skin sensitiser in this assay.