Reaction mass of Octadec-9-en-1-yl ammonium di-n-hexyl phosphorodithioate and Octadec-9-en-1-yl ammonium mono- and di-butylphosphate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was conducted between 12 June 2002 and 18 July 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

Buehler test

Justification for non-LLNA method:

An in vitro study or chemico skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available

Test material

Test material information

Constituent 1

Reference substance name:

Octadec-9-en-1-yl ammonium mono and di-butylphosphate

Molecular formula:

C22H48NO4P - C26H56NO4P

IUPAC Name:

Octadec-9-en-1-yl ammonium mono and di-butylphosphate

Constituent 2

Reference substance name:

Octadec-9-en-1-yl ammonium di-n-hexyl phosphorodithioate

Molecular formula:

C30H64NO2PS2

IUPAC Name:

Octadec-9-en-1-yl ammonium di-n-hexyl phosphorodithioate

Test material form:

liquid: viscous

Details on test material:

Material is a yellow viscous liquid

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male/female

Details on test animals and environmental conditions:

Description, Identification and Housing
Young adult, Hartley-derived albino guinea pigs were received from Hilltop Lab Animals, Inc., Scottdale, PA. Upon receipt, plastic ear tags displaying unique identification numbers were used to individually identify the animals. Cage cards displaying at least the study number, animal number and sex were affixed to each cage. The animals were housed individually in suspended stainless steel cages. All housing and care were based on the standards recommended by the Guide for the Care and Use of Laboratory Animals.

Environment
The animal room temperature and relative humidity ranges were 64-74 ° F (18-23°C) and 32-8 1 %, respectively. Environmental control equipment was monitored and adjusted as necessary to minimize fluctuations in the anim al room environment. Light timers were set to maintain a 12-hour light/ 12-hour dark cycle and room ventilation was set to produce 10- 15 air changes/hour. The room temperature and relative humidity were recorded a minimum of once daily.

Food
PMI Certified Guinea Pig Chow #5026 ( Purina Mills, Inc.) was provided ad libitum to the animals throughout the study. The lot number and expiration date of each batch of diet used during the study were recorded. The feed was analyzed and cer tified by the supplier for nutritional components and environmental contaminants. Dietary limitations for various environmental contaminants, including heavy metals, pesticides, polychlorinated biphenyls and total aflatoxin are set by the manufacturer. Within these limits, contaminants which may have been present were not expected to compromise the purpose of this study.

Results of the dietary analyses ( Certificates of Analysis) are provided by the manufacturer for each lot of diet. These are maintained by SLI.

Water
Municipal tap water treated by reverse osmosis was available ad libitum throughout the study. The purified water was supplied by an automatic watering system. Monitoring of the drinking water for contaminants is conducted by SU and the records are available for inspection. Within generally accepted limits, contaminants which may have been present were not expected to compromise the purpose of this study. The water meets the standards specified under the E PA National Drinking Water Regulations (40 CF R Part 14 1) .

Acclimation
Upon receipt, the animals were removed randomly from the shipping cartons, examined by qualified personnel, identified with plastic ear tags and then acclimated to the laboratory conditions for a minimum of five days. The animals were obse rved daily for overt physical or behavioral abnormalities, general health/moribundity and mortality.

Animal Selection
The animals chosen for study use were arbitrarily selected from healthy stock animals to avoid potential bias. All animals received a detailed pretest
obse rvation prior to dosing. Only healthy animals were chosen for study use.
Females were nulliparous and nonpregnant. The male animals were approximately 6 weeks of age and weighed 358-411 g on the day prior to Induction I dosing. The female animals were approximately 8 weeks of age and weighed 325- 351 g on the day prior to Induction I dosing.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

epicutaneous, occlusive

Vehicle:

other: Mineral oil, USP.

Induction 2

Concentration / amount:

Concentration of test material and vehicle used at induction: Induction concentrations/ test subjects: 0.5% in 80% ethanol. Concentration of test material and vehicle used for each challenge: Test subjects: 0.1% in acetone

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: Mineral oil, USP.

Challenge 2

Concentration / amount:

Concentration of test material and vehicle used at induction: Induction concentrations/ test subjects: 0.5% in 80% ethanol. Concentration of test material and vehicle used for each challenge: Test subjects: 0.1% in acetone

No. of animals per dose:

Number of animals in test group: 15
Number of animals in negative control group: 10

Details on study design:

Method of Test Article Preparation
The test article was utilized at 25% w/w in mineral oil, U S P, (Induction) and 10% w/w in mineral oil, U S P, ( Challenge/ Rechallenge). The test article preparations were prepared/dispensed fresh on each day of dosing and stirred continuously during dosing.

Study Design
This study consisted of a topical range-finding group, a test group, a challenge control group and a rechallenge control group.

Sensitization Study
Preliminary Procedures
On the day prior to each dose administration, the guinea pigs had the hair removed wit h a small animal clipper with a No. 40 blade followed by fur ther close clipping with a Braun Synco Electric Razor (Model 7507). Care was taken to avoid abrading the skin.

Dosing
A dose of 0.3 ml of the test article was placed on a 25 mm Hilltop chamber backed by adhesive tape (occlusive patch). The chambers were then applied to the clipped surface as quickly as possible.
Following chamber application, the trunk of the animal was wrapped with elastic wrap which was secured with adhesive tape to prevent removal of the chamber and the animal was returned to its cage.

Induction
On the day prior to the first induction dose administration (day -1), all test and control animals were weighed and the hair was removed from the left side of the test animals.

The induction procedure was repeated on study day 7 and on study day 14 so that a total of three consecutive induction exposures were made to the test animals.

Challenge
On the day prior to challenge dose administration, the test and challenge control animals were weighed and the hair was removed from the right side of the animals.

Rechallenge
A rechallenge was conducted in order to substantiate and clarify the challenge results. On the day prior to rechallenge dose administration, all test and rechallenge control animals were weighed and the hair was then removed from the right side of the animals.

Test Article Removal
Approximately six hours after chamber application, the binding materials were removed. The test sites were wiped with gauze moistened in mineral oil, USP , followed by dry gauze, followed by deionized water, followed by dry gauze, to remove test article residue. The animals were then returned to their cages.

Dermal Observations
The test sites were graded for irritation at approximately 24 and 48 hours following chamber application (Induction) or chamber removal (Challenge and Rechallenge) using the Dermal Grading System.

Clinical Obse rvations
Any unusual obse rvations and mortality were recorded. The animals were observed for general health/mortality twice daily, once in the morning and once in the afternoon.

Body Weights
Individual body weights were obtained for all sensitization study animals on the day prior to the first induction (day - 1) and for the appropriate test and challenge control animals on the day prior to challenge dosing. The appropriate test and rechallenge control animals were weighed on the day prior to rechallenge dosing.

Scheduled Euthanasia
All sensitization study animals were euthanized by carbon dioxide inhalation following each animal's final scoring interval. Gross necropsy examinations were not required for these animals.

Positive control substance(s):

yes

Remarks:

?-Hexylcinnamaldehyde

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

.1 %

No. with + reactions:

4

Total no. in group:

15

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: .1 %. No with. + reactions: 4.0. Total no. in groups: 15.0.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

.1 %

No. with + reactions:

1

Total no. in group:

15

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: .1 %. No with. + reactions: 1.0. Total no. in groups: 15.0.

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

.1 %

No. with + reactions:

3

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: .1 %. No with. + reactions: 3.0. Total no. in groups: 10.0.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

.1 %

No. with + reactions:

3

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: .1 %. No with. + reactions: 3.0. Total no. in groups: 10.0.

Any other information on results incl. tables

Topical Range- Finding Study

The results of the range-finding study indicated that a test article concentration of 25% w/w in mineral oil, USP, was considered appropriate for induction as the 25% concentration produced a mild to moderate dermal response. A concentration of 10% w/w in mineral oil, USP, was considered appropriate for challenge/rechallenge as it produced minimal irritation which was considered to be the highest non-irritating concentration.

 

Sensitization Study

Following challenge with 10% w/w OLOA 289 M in mineral oil, U S P, dermal scores of 1 were noted in 5/20 test animals at the 24- and 48-hour scoring intervals. Dermal reactions in the remaining test and challenge control animals were limited to scores of 0 to±.Group mean dermal scores were noted to be higher in the test animals as compared with the challenge control animals.

 

Following rechallenge with 10% w/w OLOA289 M in mineral oil, U S P, dermal scores of 1 were noted in 2/20 test animals at the 24 and 48-hour scoring intervals. Dermal reactions in the remaining test and rechallenge control animals were limited to scores of 0 to±.Group mean dermal scores were noted to be similar in the test animals as compared with the rechallenge control animals.

 

Two of the 20 (10%) induced animals responded at both challenge and rechallenge. No confounding irritation (no scores>.±) was observed at either challenge. The following animals were considered sensitized: G8658/F and G8659/ F.

 

Body Weights

The sensitization study animals gained weight during the test period and generally appeared in good health.

 

Historical Control

Using a-Hexylcinnamaldehyde (HCA) as a positive control, Springborn Laboratories, Inc., Spencerville, Ohio, has completed a study during the past six months which provided historical control data for contact sensitization to this agent utilizing the test system described herein ( Modified Buehler Design).

Following induction at 5% w/v H CA in ethanol and challenge at levels of 2.5% and 1 % w/v H CA in acetone, a contact sensitization response was observed, thereby demonstrating the susceptibility of the test system to this sensitizing agent.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information not classified Criteria used for interpretation of results: EU