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

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Number of animals: 3 animals of one sex
Age and body weight: Animals used within the study were at least 6 weeks old and body weights were at least 1 .0 kg.
Identification: earmark

Conditions
A controlled environment was maintained in the room with optimal conditions considered as being approximately 1 5 air changes per hour, a temperature of 21 ±3°C, a relative humidity of 30-70% and 12 hours artificial fluorescent light and 12 hours dark per day.

Accommodation
Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm). Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.

Diet
Standard laboratory rabbit diet {Teklad Global Rabbit Diet, code 2030, Harlan, Blackthorn, England) approx. 1 00 g. per day. Certificates of analysis were examined and retained in the NOTOX archives. In addition, hay (BMI, Helmond, the Netherlands) was provided twice a week.

Water
Free access to tap-water. Certificates of quarterly analysis were examined and retained in the NOTOX archives.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 ml

Duration of treatment / exposure:

Sinngle application

Observation period (in vivo):

Single application. Total observation period of 14 days post instillation.

Number of animals or in vitro replicates:

3

Details on study design:

Since marked effects were considered possible and at request of the sponsor this eye irritation study was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 1 4 days later, after considering the degree of eye irritation observed in the first animal.

A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the eyes, which were free from any abnormality.

Each animal was treated by instillation of 0 .1 ml of the test substance in the conjunctiva! sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control.

Immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7 .0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. When considered necessary, this procedure was repeated to assess recovery. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

316 - Sentinel

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 hrs

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

Animal # 355

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 hrs

Results 3

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

Animals # 356

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hrs

Results 4

Irritation parameter:

iris score

Basis:

mean

Remarks:

316 - Sentinel

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 72 hrs

Results 5

Irritation parameter:

iris score

Basis:

mean

Remarks:

Animal #355

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

0.67

Max. score:

2

Reversibility:

not fully reversible within: 14 days

Results 6

Irritation parameter:

iris score

Basis:

mean

Remarks:

Animal #356

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

0

Max. score:

2

Remarks on result:

other: Effects noted at 1 hr, but not observed at 24 hrs and subsequent to that.

Results 7

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Remarks:

316 - Sentinel

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

2.67

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 8

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Remarks:

Animal #355

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

2.33

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 9

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Remarks:

Animal #356

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

2.33

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 10

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

316 - Sentinel

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 11

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

Animal #355

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 12

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

Animal #356

Time point:

other: Mean of scores at 24, 48 & 72 hrs

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 72 hrs

Irritant / corrosive response data:

Irritation
Instillation of 0 .1 ml of the test substance into one eye of each of three rabbits resulted in effects on the cornea, iris and conjunctivae.

Corneal injury was seen as opacity (maximum grade 1 ) and epithelial damage (maximum 25 or 50 % of the corneal area). The corneal injury had resolved within 72 hours in two animals and within 7 days in the other animal.

lridial irritation (grade 1 ) was observed which had resolved within 24 or 72 hours or within 7 days.

Irritation of the conjunctivae was seen as redness, chemosis and discharge, which had completely resolved within 1 4 days in all animals.

Corrosion
There was no evidence of ocular corrosion.

Other effects:

Colouration I Remnants
No staining of (peri) ocular tissues by the test substance was observed.

oxicity I Mortality
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

Category 2A (irritating to eyes)

Remarks:

Migrated information Criteria used for interpretation of results: EU