Reaction mass of 9-icosyl-9-phosphabicyclo[3.3.1]nonane and 9-icosyl-9-phosphabicyclo[4.2.1]nonane

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

9 February 1967 and 26 May 1967

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1967

Report date:

1967

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: The intracutaneous injection procedure employed was modeled upon that described by Draize*.

Deviations:

not specified

Principles of method if other than guideline:

* "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. " Published by The Editorial Committee, Association of Food and Drug
Officials of the United States (1959). Business Office, Bureau of Food and Drugs, Texas State Department of Health, Austin 1, Texas.
See chapter entitled "Dermal Toxicity" by John H. Draize, p. 46 et seq.

GLP compliance:

no

Remarks:

Predates GLP

Type of study:

Draize test

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 9-icosyl-9-phosphabicyclo[3.3.1]nonane and 9-icosyl-9-phosphabicyclo[4.2.1]nonane

EC Number:

915-206-4

Molecular formula:

C28H55P

IUPAC Name:

Reaction mass of 9-icosyl-9-phosphabicyclo[3.3.1]nonane and 9-icosyl-9-phosphabicyclo[4.2.1]nonane

Details on test material:

- Name of test material (as cited in study report): RM-17
- Substance type: No data
- Physical state: No data
- Analytical purity: No data
- Impurities (identity and concentrations): No data
- Purity test date: No data
- Lot/batch No.: No data
- Expiration date of the lot/batch: No data
- Stability under test conditions: No data
- Storage condition of test material: No data

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: Albino

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Not stated.
- Age at study initiation: Not stated.
- Weight at study initiation: Weight range 300 to 500 grams.
- Housing: The animals were housed in standard laboratory cages.
- Diet: ad libitum; Commercial stock guinea pig ration, supplemented with fresh cabbage.
- Water: ad libitum
- Acclimation period: Not stated.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not stated.
- Humidity (%): Not stated.
- Air changes (per hr): Not stated.
- Photoperiod (hrs dark / hrs light): Not stated.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

other: intracutaneous

Vehicle:

propylene glycol

Induction 2

Concentration / amount:

The solution employed for the injections was 0.10 per cent (w/v) RM-17 in propylene glycol.

Challenge

Challenge 1

Route:

other: intracutaneous

Vehicle:

propylene glycol

Challenge 2

Concentration / amount:

The solution employed for the injections was 0.10 per cent (w/v) RM-17 in propylene glycol.

No. of animals per dose:

Induction
Ten male guinea pigs received a total of ten injections each.
Challenge
Eight remaining guinea pigs received one injection each.*

* Two of the animals died during the course of the study. The two deaths were not thought to be associated with administration of test material.

Details on study design:

RANGE FINDING TESTS: No data

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: Ten exposures per animal
- Exposure period: A series of "sensitizing" injections were made intracutaneously on Mondays, Wednesdays and Fridays until each pig had received a total of ten such injections.
- Control group: no
- Site: On the day before initiating the experiment, the backs· and flanks of each animal were clipped free of hair with electric clippers using care to avoid injuring the integument. Thereafter, clipping of each guinea pig was done immediately prior to giving each injection in the specified series.
- Concentrations: The solution employed for the injections was 0.10 per cent (w/v) test material in propylene glycol.
A volume of 0.05 mL was used for the first injection.
The remaining injections (numbers 2 through 10 of the "sensitizing" injections) were 0.1 mL each.
All injections were made using a one-cc tuberculin syringe fitted with a 26-gauge hypodermic needle.

B. CHALLENGE EXPOSURE
- No. of exposures: One retest or challenge injection was made into each animal.
- Day(s) of challenge:
- Exposure period: Two weeks after the tenth induction injection.
- Control group: no
- Site: Injections were made at random into an area three to four centimeters square on the back and upper flanks of each animal.
- Concentrations: The solution employed for the injections was 0.10 per cent (w/v) test material in propylene glycol.
A volume of 0.05 mL was used for the challenge injection.

- Evaluation (hr after challenge): Twenty-four hours after each injection the diameter (for a circular reaction site) or the major axis (of an elliptical reaction site) was measured and recorded for each individual reaction. In addition, both erythema and edema were graded separately on scales of 0 to 4 using the system described by Draize for the scoring of skin irritation.

Challenge controls:

No data.

Positive control substance(s):

not specified

Results and discussion

Positive control results:

Not applicable.

In vivo (non-LLNA)

Results

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0.10 per cent (w/v)

Total no. in group:

10

Clinical observations:

Challenge responses were well within the statistical limits set by the responses to the sensitizing injections.

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.10 per cent (w/v). Total no. in groups: 10.0. Clinical observations: Challenge responses were well within the statistical limits set by the responses to the sensitizing injections..

Any other information on results incl. tables

The results of the skin sensitization study on the test material are surnmarized in Tables I, II, and III below. In these tables the reaction site data following the challenge injection are compared with those data for the ten sensitizing injections on an animal- by-animal basis.

Examination of these data disclosed no evidence of a challenge response in a given animal which differed significantly from that which would be predictable from the mean data accumulated for the responses to the sensitizing injections for that animal. Challenge responses were well within the statistical limits set by the responses to the sensitizing injections.

Table I: Statistics for Major Axes (Diameters) of Reaction Sites

Length of Major Axes (Diameters) of Reaction Sites
Values for Ten Sensitizing Injections
Animal Number	Mean (cm)	Range (cm)	Estimated Standard Deviation (cm)	Challenge Injection Value for Individual Animal (cm)
1	1.16	1.08 to 1.27	0.06	1.15
2	1.14	1.05 to 1.28	0.08	1.19
3	1.17	1.02 to 1.30	0.09	1.18
4	1.19	1.10 to 1.35	0.08	1.09
5	1.15	0.95 to 1.30	0.11	1.08
6	1.22	1.12 to 1.30	0.06	1.25
7	1.14	1.01 to 1.25	0.08	1.10
8	1.21	1.10 to 1.34	0.08	1.20

Table II: Statistics for Erythema Scores of Reaction Sites

Degree of Erythema at Reaction Sites
Values for Ten Sensitizing Injections
Animal Number	Mean	Range	Estimated Standard Deviation	Challenge Injection Value for Individual Animal
1	2.6	2 to 3	0.3	3
2	2.4	2 to 3	0.3	3
3	2.7	2 to 3	0.3	3
4	2.9	2 to 3	0.3	3
5	2.5	2 to 3	0.3	3
6	2.9	2 to 3	0.3	3
7	2.7	2 to 3	0.3	3
8	3.0	None	0.0	3

Table III: Statistics for Edema Scores of Reaction Sites

Degree of Edema at Reaction Sites
Values for Ten Sensitizing Injections
Animal Number	Mean	Range	Estimated Standard Deviation	Challenge Injection Value for Individual Animal
1	1.4	1 to 2	0.3	1
2	1.2	1 to 2	0.3	1
3	1.4	1 to 3	0.6	1
4	1.6	1 to 3	0.6	1
5	1.5	1 to 3	0.6	1
6	1.5	1 to 2	0.3	1
7	1.4	1 to 2	0.3	2
8	1.7	1 to 3	0.6	1

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

It was concluded from these data that evidence for significant skin sensitizing properties of the test material was lacking.

Executive summary:

A number of toxicological studies were requested on the test material, among which was an evaluation of its potential skin sensitizing properties.

Accordingly, the guinea pig intracutaneous injection technique (as described by Draize), a procedure useful in "screening out" potentially strong or severe skin sensitizing agents, was conducted on the test material as a 0. 10 per cent (w/v) solution in propylene glycol.

The results of this test provided no evidence that the test material could produce skin sensitization in the guinea pig.