Benzenepropanamine, 2-methoxy-5-methyl-N,N-bis(1-methylethyl)-.gamma.-phenyl-, hydrobromide (1:1)

Endpoint summary

Description of key information

Key study: Skin irritation: OECD guideline 404 and EU method B.4. GLP study.
The test item produced a primary irritation index of 0.0 and was classified as non-irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.
Key study: Eye irritation: OECD guideline 405 and EU method B.5. GLP study. 
The persistence of reactions in one treated eye at the 21-Day observation was considered to be indicative of irreversible ocular damage. One treated eye appeared normal at the 7-Day observation. The test item produced irreversible ocular damage in one animal and was considered to be CORROSIVE to the rabbit eye.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline and EU method. GLP study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Leicestershire, UK
- Age at study initiation: Approximately 12 - 20 weeks at the start of treatment
- Weight at study initiation: 2.49-3.14 kg
- Housing: lndividually housed in suspended metal and/or plastic cages.
- Diet (e.g. ad libitum): 2930 Teklad Global Certified Diet, at libitum
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23 ºC
- Humidity (%): 30 to 70%
- Air changes (per hr): At least 15 changes per hour
- Photoperiod (hrs dark / hrs light): Twelve hours of continuous artificial light in each twenty-four hour period

Type of coverage:

semiocclusive

Preparation of test site:

other: On the day before treatment the back and flanks of each animal will be clipped free of fur using veterinary clippers.

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g moistened sufficiently with 0.5 ml of distilled water to achieve a paste.

Duration of treatment / exposure:

Animal 1: 3 minutes, 1 hour and 4 hours
Animal 2: 4 hours

Observation period:

Immediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation.

Number of animals:

Two animals

Details on study design:

TEST SITE
- Area of exposure: 6 cm2
- Type of wrap if used: The test item will be applied to a small area of skin (approximately 6 cm2) under a cotton gauze patch (2.5 cm x 2.5 cm). The patch will be secured to the skin using a single strip of surgical adhesive tape (approximate size 2.5 cm x 4.0 cm). To prevent interference with the patches and ingestion of the test item the animals will be fitted with an elasticated corset for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Any residual test item was removed by gentle swabbing with cotton wool soaked in distilled water.

SCORING SYSTEM:
Erythema and Eschar formation:
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness) to eschar formation preventing grading of erythema 4

Oedema Formation:
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well defined by definite raising) 2
Moderate oedema (raised approximately 1 millimetre) 3
Severe oedema (raised more than 1 millimetre extending beyond the area of exposure) 4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Irritant / corrosive response data:

3-Minute Exposure Period
No evidence of skin irritation was noted during the study.
1-Hour Exposure Period
No evidence of skin irritation was noted during the study.
4-Hour Exposure Period
No evidence of skin irritation was noted during the study.

Other effects:

Both animals showed expected gain in bodyweight during the study.

Table 1: Individual Skin Reactions Following 4-Hour Exposure

	24 h		48 h		72 h		Mean 24-72 h
	Animal No.		Animal No.		Animal No.
	1	2	1	2	1	2
Erythema	0	0	0	0	0	0	0
Oedema	0	0	0	0	0	0	0

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item produced a primary irritation index of 0.0 and was classified as non-irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

Executive summary:

The study was performed to assess the irritancy potential of the test item to the skin of the New Zealand White rabbit. The method was designed to be compatible with the following:

- OECD Guidelines for the Testing of Chemicals No. 404 “Acute Dermal Irritation/Corrosion” (adopted 24 April 2002)

. Method B4 Acute Toxicity (Skin Irritation) of Commission Regulation (EC) No. 440/2008

3-minute and 1-hour semi-occluded applications of the test item to the intact skin of one rabbit produced no evidence of skin irritation.

A single 4-hour, semi-occluded application of the test item to the intact skin of two rabbits produced no evidence of skin irritation.

The test item produced a primary irritation index of 0.0 and was classified as non-irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

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: OECD guideline and EU method. GLP study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Leicestershire, UK
- Age at study initiation: Approximately 12 - 20 weeks at the start of treatment
- Weight at study initiation: 2.11 or 2.50 kg
- Housing: lndividually housed in suspended metal and/or plastic cages.
- Diet (e.g. ad libitum): 2930 Teklad Global Certified Diet, at libitum
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23 °C
- Humidity (%): 30 to 70%
- Air changes (per hr): At least 15 changes per hour
- Photoperiod (hrs dark / hrs light): Twelve hours of continuous artificial light in each twenty-four hour period

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eye remained untreated and was used for control purposes.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): A volume of 0.1 ml of the test item, which was found to weigh approximately 75 mg (as measured by gently compacting the required volume into an adapted syringe).

Duration of treatment / exposure:

24 h

Observation period (in vivo):

Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment. Additional observations were made on Days 7, 14 and 21 to assess the reversibility of the ocular effects.

Number of animals or in vitro replicates:

Two animals

Details on study design:

SCORING SYSTEM:
1. CONJUNCTIVAE
Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Vessels normal 0
Vessels definitely injected above normal 1
More diffuse, deeper crimson red, individual vessels not easily discernible 2
Diffuse beefy red 3
Chemosis
No swelling 0
Any swelling above normal (includes nictitating membrane) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids half closed to completely closed 4
Discharge
No discharge 0
Any amount different from normal (does not include small amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs a considerable area around the eye 3
2. IRIS
Values
Normal 0
Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or 1
combination of any thereof) iris still reacting to light (sluggish reaction is positive)
No reaction to light, haemorrhage, gross destruction (any or all of these) 2

3. CORNEA
Degree of Opacity (most dense area used)
No opacity 0
Scattered or diffuse areas, details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Opalescent areas, no details of iris visible, size of pupil barely discernible 3
Opaque, iris not discernible through the opacity 4
Area of Cornea Involved
One quarter (or less) but not zero 1
Greater than one quarter but less than half 2
Greater than half but less than three quarters 3
Greater than three quarters, up to whole area 4

TOOL USED TO ASSESS SCORE: Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Time point:

other: 24-72 h

Score:

1.33

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

1.33

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

2

Max. score:

2

Reversibility:

not reversible

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 21 days

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Time point:

other: 24-72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

3

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

No corneal effects were noted in one treated eye during the study. Iridial inflammation was noted in this treated eye one hour after treatment. Moderate conjunctival irritation was noted in this treated eye one and 24 hours after treatment with minimal conjunctival irritation noted at the 48 and 72-Hour observations. This treated eye appeared normal at the 7-Day observation.

Translucent corneal opacity was noted in the other treated eye throughout the study. Iridial inflammation was noted in this treated eye one hour after treatment and at the 24, 48, 72-Hour, 7 and 14-Day observations. Severe conjunctival irritation was noted in this treated eye one hour after treatment and at the 24 and 48-Hour observations with moderate conjunctival irritation noted at the 72-Hour and 7-Day observations. Haemorrhage on the upper and lower nictitating membranes was noted in this treated eye at the 72-Hour observation. A white area on the cornea, approximately 2 mm x 7 mm in size, was noted in this treated eye at the 7 and 14-Day observations. Vascularisation, with a generalised ingrowth of vessels approximately 3 mm in size, was noted in this treated eye at the 14 and 21-Day observations. The persistence of reactions in this treated eye at the 21-Day observation was considered to be indicative of irreversible ocular damage.

Other effects:

Both animals showed expected gains in bodyweight during the study.

Table 1 Individual Scores for Ocular Irritation

Rabbit Number and Sex	72207 Male					72261 Male
Time After Treatment	1 Hour	24 Hours	48 Hours	72 Hours	7 Days	1 Hour	24 Hours	48 Hours	72 Hours	7 Days	14 Days	21 Days
CORNEA
Degree of Opacity	0	0	0	0	0	2	2	2	2	2 x	2 x V	2V
Area of Cornea Involved	0	0	0	0	0	3	3	4	4	4	4	4
IRIS	1	0	0	0	0	1	1	1	1	1	1	0
CONJUNCTIVAE
Redness	2	2	1	1	0	2	2	2	2H	2	0	0
Chemosis	2	2	1	1	0	3	3	3	3	2	0	0
Discharge	2	2	1	0	0	3	3	3	2	1	0	0
H = Areas of haemorrhage on the upper and lower nictitating membranes V = Vascularisation, with a generalised ingrowth of vessels approximately 3 mm in size x = A white area on the cornea, approximately 2 mm x 7 mm in size

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The persistence of reactions in one treated eye at the 21-Day observation was considered to be indicative of irreversible ocular damage. One treated eye appeared normal at the 7-Day observation. The test item produced irreversible ocular damage in one animal and was considered to be CORROSIVE to the rabbit eye.

Executive summary:

The study was performed to assess the irritancy potential of the test item to the eye of the New Zealand White rabbit. The method was designed to be compatible with the following:

-OECD Guidelines for the Testing of Chemicals No. 405 “Acute Eye Irritation/Corrosion” (adopted 24 April 2002)

-Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008

A single application of the test item to the non-irrigated eye of two rabbits produced translucent corneal opacity, iridial inflammation and moderate to severe conjunctival irritation. Other ocular effects noted in one treated eye were a white area on the cornea, vascularisation, with a generalised ingrowth of vessels and areas of haemorrhage on the upper and lower nictitating membranes. The persistence of reactions in one treated eye at the 21-Day observation was considered to be indicative of irreversible ocular damage. One treated eye appeared normal at the 7-Day observation. The test item produced irreversible ocular damage in one animal and was

considered to be CORROSIVE to the rabbit eye.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Key study: Skin irritation: OECD guideline 404 and EU method B.4. GLP study.

The test item produced a primary irritation index of 0.0 and was classified as non-irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

Key study: Eye irritation: OECD guideline 405 and EU method B.5. GLP study.

The persistence of reactions in one treated eye at the 21-Day observation was considered to be indicative of irreversible ocular damage. One treated eye appeared normal at the 7-Day observation. The test item produced irreversible ocular damage in one animal and was considered to be CORROSIVE to the rabbit eye.

Justification for selection of skin irritation / corrosion endpoint:
Only one study available

Justification for selection of eye irritation endpoint:
Only one study available

Effects on eye irritation: corrosive

Justification for classification or non-classification

Based on the available data, the substance is classified as Eye damage Category 1.