Insulin DesB30

Endpoint summary

Description of key information

Data from OECD 404 guideline studies on MI3 and X14DesB30 indicate no potential for skin irritation. Due to very close structural similarities a lack of skin irritation potential can be concluded for Insulin DesB30 as well.

Data from OECD 405 guideline studies on MI3 and X14DesB30 indicate no potential for eye irritation. Due to very close structural similarity to Insulin DesB30

a lack of eye irritation potential can be concluded for Insulin DesB30 as well.

For further read-across justification see document attached to section 13.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Interpretation of results:

GHS criteria not met

Conclusions:

No data is available for Insulin DesB30 (T).
Data from OECD 404 guideline studies on MI3 and X14DesB30 indicate no potential for skin irritation. Due to very close structural similarities, a lack of skin irritation potential can be concluded for Insulin DesB30 as well.
See justification for read-across attached in section 13.

Executive summary:

No data is available for Insulin DesB30 (T).

Data from OECD 404 guideline studies on MI3 and X14DesB30 indicate no potential for skin irritation. Due to very close structural similarities a lack of skin irritation potential can be concluded for Insulin DesB30 as well.

See justification for read-across attached in section 13.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000.04.04 - 2000.07.04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

Storage temperature of samples, the humidity in the animal holding room, and the time of recording the dermal reactions deviated from what stated in the protocol. None of the deviation is considered to have affected the study outcome.

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Storage temperature of samples, the humidity in the animal holding room, and the time of recording the dermal reactions deviated from what stated in the protocol. None of the deviation is considered to have affected the study outcome.

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

yes

Remarks:

Storage temperature of samples, the humidity in the animal holding room, and the time of recording the dermal reactions deviated from what stated in the protocol. None of the deviation is considered to have affected the study outcome.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL (MI3 dried)
- Source and lot/batch No.of test material: C202483
- Expiration date of the lot/batch: 2001.05.31
- Purity: 98.4%
- Purity test date: 2001.05.31

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: < -10 degrees Celcius
- Stability under test conditions: NA
- Solubility and stability of the test substance in the solvent/vehicle: Soluble in vehicle (water)
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: NA

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Purified water (Elgastat purifier used)
- Final preparation of a solid: 100mg/ml MI3 in water, pH of 8.0.

FORM AS APPLIED IN THE TEST (if different from that of starting material): solid dissolved in purified water.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Rabbit (New Zealand White rabbit)
- Sex: male and female
- Age at study initiation: 10 - 14 weeks.
- Weight at study initiation: 2.35 - 2.41 kg
- Housing: Single housed in floor-pens with wood chips bedding
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: NA

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-22
- Humidity (%): 40 – 80 %
- Air changes (per hr): at least 10 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours/12 hours

IN-LIFE DATES: From: To: at least 10 - 13 days

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.5 mL of MI3 moistened with water (paste)

Duration of treatment / exposure:

4 hours

Observation period:

one hour, 24, 48 and 72 hours after removal of patch and dressing.

Number of animals:

3 ( one male and two females)

Details on study design:

TEST SITE
- Area of exposure: 30 x 20 mm
- % coverage: NA
- Type of wrap if used: Dense elastic gauze bandage (semi-occlusive)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Light brushing and swabbing with moistened cotton wool
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) one hour, 24, 48 and 72 hours after application.

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No indication of irritation or corrosive response. Individual and group mean values for dermal changes ( Erythema and Oedema) in the tested animals were zero at 0, 24, 48 and 72 hours.

Other effects:

No other effects observed

Interpretation of results:

GHS criteria not met

Conclusions:

The Study was conducted to determine the irritation caused to intact rabbit skin following a single (4 hour), semi-occluded, topical application of MI3. The study was designed in accordance with OECD guideline method 404. One male and two female rabbits were exposed to a single dose of 0.5mL MI3 applied to 30 x 20 mm area.All animals were killed by an intravenous injection of an overdose of barbiturate at day 4. No reactions were observed following a single semi-occluded, topical application of MI3 to intact rabbit skin for four hours.

Executive summary:

The Study was conducted to determine the irritation caused to intact rabbit skin following a single (4 hour), semi-occluded, topical application of MI3. The study was designed in accordance with OECD guideline method 404, the commission directive 92/69/EEC, Method B4, and the US EPA Health effects test Guidelines OPPTS 870.2500. A single dose of 0.5 mL MI3 that was applied to a 30 x 20 mm area on the clipped dorsum of each of three rabbits, one male and two females. The treated area of skin was covered by a semi- occlusive bandage for 4 hours and assessed for dermal reaction the following three days. All animals were killed by an intravenous injection of an overdose of barbiturate at day 4.

In conclusion, no dermal reactions were observed following a single semi-occluded, topical application of MI3 to intact rabbit skin for four hours.

Reference 3

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000.03.31 - 2000.07.04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

The relative humidity in the animal holding room was consistently lower (15 %) than stated in the protocol. The sentinel rabbit assessments were conducted on Day 9 rather than Day 8. The deviations are not considered to compromise the study integrity

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

The relative humidity in the animal holding room was consistently lower (15 %) than stated in the protocol. The sentinel rabbit assessments were conducted on Day 9 rather than Day 8. The deviations are not considered to compromise the study integrity

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

yes

Remarks:

The relative humidity in the animal holding room was consistently lower (15 %) than stated in the protocol. The sentinel rabbit assessments were conducted on Day 9 rather than Day 8. The deviations are not considered to compromise the study integrity

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL (X14DesB30)
- Source and lot/batch No.of test material: X1YIS0201014
- Expiration date of the lot/batch: NA
- Purity 80%
- Purity test date: 2000.02.18

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: < -70 degrees Celcius
- Stability under test conditions: NA
- Solubility and stability of the test substance in the solvent/vehicle: Soluble in vehicle (water)
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: NA

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Purified water.
- Final preparation of a solid: A 50% m/v dispersion in purified water, pH of 3.22.

FORM AS APPLIED IN THE TEST (if different from that of starting material): A 50% m/v dispersion in purified water.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Rabbit (New Zealand White rabbit)
- Sex: male and female
- Age at study initiation: 11 - 13 weeks
- Weight at study initiation: 2.35 - 2.41 kg
- Housing: Single housed in floor-pens with wood chips bedding
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: NA

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-22
- Humidity (%): 40 – 80 %
- Air changes (per hr): at least 10 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours/12 hours

IN-LIFE DATES: From: To: at least 17 days

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Remarks:

50% m/v dispersion, pH of 3.22

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5mL X14DesB30 as a paste

Duration of treatment / exposure:

four hours

Observation period:

One hour, 24, 48 and 72 hours

Number of animals:

3 (one male and two females)

Details on study design:

TEST SITE
- Area of exposure: 30 x 20 mm
- % coverage: NA
- Type of wrap if used: Dense elastic gauze bandage (semi-occlusive)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Light brushing and swabbing with moistened cotton wool
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) one hour, 24, 48 and 72 hours after application.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial haemorrhage (transient - recovery after 4 days) present after removal of dose residuals.

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial hemorrhage (transient - recovery after 24h) present after removal of dose residuals. Developed scabs by Day 3 and showed full resolution by Day 4.

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial hemorrhage (transient - recovery after 24h) present after removal of dose residuals. Developed scabs by Day 3 and showed full resolution by Day 4.

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial haemorrhage (transient - recovery after 4 days) present after removal of dose residuals.

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial hemorrhage (transient - recovery after 24h) present after removal of dose residuals. Developed scabs by Day 3 and showed full resolution by Day 4.

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial hemorrhage (transient - recovery after 24h) present after removal of dose residuals. Developed scabs by Day 3 and showed full resolution by Day 4.

Interpretation of results:

GHS criteria not met

Conclusions:

The Study was conducted to determine the irritation caused to intact rabbit skin following a single (4 hour), semi-occluded, topical application of X14DesB30. The study was designed in accordance with OECD guideline method 404. One male and two female rabbits were exposed to a single dose of 0.5 mL X14DesB30 applied to a 30 x 20 mm area on the clipped dorsum. All animals were killed by an intravenous injection of an overdose of barbiturate at day 4. X14DesB30 did not cause any notable dermal reactions. All observed reactions were a result of mechanical trauma caused by removal of the dose residuals.

Executive summary:

The Study was conducted to determine the irritation caused to intact rabbit skin following a single (4 hour), semi-occluded, topical application of X14DesB30. The study was designed in accordance with OECD guideline method 404, the commission directive 92/69/EEC, Method B4, and the US EPA Health effects test Guidelines OPPTS 870.2500. A single dose of 0.5 mL X14DesB30 was applied to a 30 x 20 mm area on the clipped dorsum of each of three rabbits, one male and two female rabbits. A semi- occlusive bandage covered the treated area of skin for 4 hours and dermal reactions to treatment were assessed for up to nine days after removal of the dressings. All animals were killed by an intravenous injection of an overdose of barbiturate at Day four or Day nine. Transient petechial hemorrhage was observed as a result of mechanical trauma caused by removal of the dose residuals. Summarized, X14DesB30 did not cause any notable dermal reactions and classified not to cause skin irritation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Remarks:

S2 and S3

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Remarks:

S2 and S3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 0.1

Max. score:

0.3

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Remarks:

S2

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 0.1

Max. score:

0.3

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Remarks:

S3

Interpretation of results:

GHS criteria not met

Conclusions:

No data is available for Insulin DesB30 (T).
Data from OECD 405 guideline studies on MI3 and X14DesB30 indicate no potential for eye irritation. Due to very close structural similarity to Insulin DesB30, a lack of eye irritation potential can be concluded for Insulin DesB30 as well.
See further read-across documentation attached in section 13.

Executive summary:

No data is available for Insulin DesB30 (T).

Data from OECD 405 guideline studies on MI3 and X14DesB30 indicate no potential for eye irritation. Due to very close structural similarity to Insulin DesB30, a lack of eye irritation potential can be concluded for Insulin DesB30 as well.

See further read-across documentation attached in section 13.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000.03.07 - 2000.07.04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

The nominal temperature of the freezer was kept at < -10oC and not <-20 oC as, and the relative humidity in the animal holding room was up to 15% lower than stated in the protocol. The deviations are not considered to compromise the study integrity

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

The nominal temperature of the freezer was kept at < -10oC and not <-20 oC as, and the relative humidity in the animal holding room was up to 15% lower than stated in the protocol. The deviations are not considered to compromise the study integrity

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

yes

Remarks:

The nominal temperature of the freezer was kept at < -10oC and not <-20 oC as, and the relative humidity in the animal holding room was up to 15% lower than stated in the protocol. The deviations are not considered to compromise the study integrity

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL (MI3 dried)
- Source and lot/batch No.of test material: C202483
- Expiration date of the lot/batch: 2001.05.31
- Purity: 98.4%
- Purity test date: 2001.05.31

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: < -10 degrees Celcius
- Stability under test conditions: NA
- Solubility and stability of the test substance in the solvent/vehicle: Soluble in vehicle (water)
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: NA

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Purified water (Elgastat purifier used)
- Final preparation of a solid: 100mg/ml MI3 in water, pH of 8.0.

FORM AS APPLIED IN THE TEST (if different from that of starting material): solid dissolved in purified water.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Rabbit (New Zealand White rabbit)
- Sex: male and female
- Age at study initiation: 11 - 14 weeks.
- Weight at study initiation: 1.97 - 2.54 kg
- Housing: Single housed in floor-pens with wood chips bedding
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: NA

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-22
- Humidity (%): 40 – 80 %
- Air changes (per hr): at least 10 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours/12 hours

IN-LIFE DATES: From: To: at least 14 - 16 days

Vehicle:

water

Controls:

other: The unexposed eye of the same animal served as a untreated control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

Duration of treatment / exposure:

single instillation dose into the left conjunctival

Observation period (in vivo):

Observations were recorded at: immediately after dosing, 30 min, one hour after dosing and at least twice within the first 24 hours after dosing. Thereafter observations were made once a day up to the third day after exposure.
However, the observation period was extended for the sentinel to five days to allow resulution of minor iridial changes prior to dosing of the remaining animals.

Number of animals or in vitro replicates:

3 animals

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): NA
- Time after start of exposure: NA

SCORING SYSTEM: Standard grading scheme (SGS) graded 0 to 4 (where 0 is no response and 4 is the most severe response).
The following endpoints were evaluated/scored (Sting repsonse; Cornea - degree of opacity; Cornea -area of corneal opacity; Iris; Conjunctivae- redness; Conjunctivae - chemosis, and Conjunctivae -discharge)


TOOL USED TO ASSESS SCORE:fluorescein and ultraviolet light.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 0.1

Max. score:

0.3

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

0

Remarks on result:

no indication of irritation

Other effects:

- Lesions and clinical observations: No indications of systemic toxicity or ill health were noted for any of the rabbits during the study. MI3 caused practically no initial sting reaction in two animals and a moderate sting in the third.
- Ophthalmoscopic findings: A darkned/red area appeared aorund the iris approximately four hours after instillation in the sentinel animal, but had disappeared by the 24 hour assessment. However, a residual dark speck in the iris appeared from Day 3 to Day 5, and disappeared at Day 6. It is not known if this is a direct result of the indtillation procedure, but it did not apear in the two other animals.
- Histopathological findings: NA
- Effects of rinsing or washing: NA
- Other observations: the eyes of all rabbits were overtly normal by the 48 hour examination with the exception of the persistant dark speck in the iris of the sentinel rabbit until Day 6.

Interpretation of results:

GHS criteria not met

Conclusions:

The Study was conducted to determine the irritation caused to the rabbit eye following a single instillation of MI3 into the conjunctival sac. The study was designed in accordance with OECD guideline method 405. One male and two female rabbits received a single instillation of 0.1 mL MI3 into the conjunctival sac of the eye. There was no indicative of systemic toxicity or ill health for any of the rabbits. The instillation caused slight changes in the conjunctivae that resolved within 24 or 48-hours. An exception was a persistent dark speck in the edge of the iris of the sentinel rabbit only. The cornea of each eye was overtly unaffected.

Executive summary:

The Study was conducted to determine the irritation caused to the rabbit eye following a single instillation of MI3 into the conjunctival sac. The study was designed in accordance with OECD guideline method 405, the commission directive 92/69/EEC, Method B5, and the US EPA Health effects test Guidelines OPPTS 870.2400. One male and two female rabbits received a single instillation of 0.1 mL MI3 into the conjunctival sac of the eye. There was no indicative of systemic toxicity or ill health for any of the rabbits. The instillation caused practically no initial sting reaction in two animals and a moderate sting reaction in the third. Only minimal effects were observed after instillation of MI3 to eyes of all three animals. These effects were a slight reddening of the conjunctivae, a slight injection of the conjunctival vasculature, and production of a very slight discharge which all disappeared after 24 or 48-hours. An exception was a persistent dark speck in the edge of the iris of the sentinel rabbit only. The cornea of each eye was overtly unaffected.

Reference 3

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:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

The relative humidity in the animal holding room fell below (up to 15%) the humidity range 40 – 80% RH stated in the protocol. This deviation was not considered to have compromised the study integrity.

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

The relative humidity in the animal holding room fell below (up to 15%) the humidity range 40 – 80% RH stated in the protocol. This deviation was not considered to have compromised the study integrity.

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

yes

Remarks:

The relative humidity in the animal holding room fell below (up to 15%) the humidity range 40 – 80% RH stated in the protocol. This deviation was not considered to have compromised the study integrity.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL (X14DesB30, dried)
- Source and lot/batch No.of test material: Y1YIS201014
- Expiration date of the lot/batch: NA
- Purity: 80%
- Purity test date: 2000.02.18

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: < -70 degrees Celsius
- Stability under test conditions: NA
- Solubility and stability of the test substance in the solvent/vehicle: Soluble in vehicle (water)
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: NA

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Purified water (Elgastat purifier used)
- Final preparation of a solid: 50 % m/v dispersion in water, pH of 3.22.

FORM AS APPLIED IN THE TEST (if different from that of starting material): solid dissolved in purified water.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Rabbit (New Zealand White rabbit)
- Sex: male and female
- Age at study initiation: 11 - 14 weeks.
- Weight at study initiation: 2.15 to 2.45 kg
- Housing: Single housed in floor-pens with wood chips bedding
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: NA

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-22
- Humidity (%): 40 – 80 %
- Air changes (per hr): at least 10 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours/12 hours

IN-LIFE DATES: From: To: 11 - 14 days

Vehicle:

water

Controls:

other: The untreated eye served as a control for the treated eye.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

Duration of treatment / exposure:

single instillation dose into the left conjunctival

Observation period (in vivo):

Observations were recorded at; immediately after dosing, 30 min, one hour after dosing and at least twice within the first 24 hours after dosing. Thereafter observations were made at least once a day up to the third day after exposure.

Number of animals or in vitro replicates:

3 animals

Details on study design:

RREMOVAL OF TEST SUBSTANCE
- Washing (if done): NA
- Time after start of exposure: NA

SCORING SYSTEM: Standard grading scheme (SGS) graded 0 to 4 (where 0 is no response and 4 is the most severe response).
The following endpoints were evaluated/scored (Sting repsonse; Cornea - degree of opacity; Cornea -area of corneal opacity; Iris; Conjunctivae- redness; Conjunctivae - chemosis, and Conjunctivae -discharge).

TOOL USED TO ASSESS SCORE:fluorescein and ultraviolet light.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.1

Max. score:

0.3

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Other effects:

- Lesions and clinical observations: No indicative of systemic toxicity or ill health.

Interpretation of results:

GHS criteria not met

Conclusions:

The study was designed in accordance with OECD guideline method 405. One male and two female rabbits received a single instillation of 0.1 mL X14DesB30 into the conjunctival sac of the eye. There was no indicative of systemic toxicity or ill health for any of the rabbits. The instillation caused in all three rabbits slight changes in the conjunctivae and iris. The cornea of each eye was overtly unaffected and all reactions had overtly resolved within 48 hours of instillation.

Executive summary:

The study was designed in accordance with OECD guideline method 405, the commission directive 92/69/EEC, Method B5, and the US EPA Health effects test Guidelines OPPTS 870.2400. One male and two female rabbits received a single instillation of 0.1 mL X14DesB30 into the conjunctival sac of the eye. There was no indicative of systemic toxicity or ill health for any of the rabbits. The instillation caused practically no initial sting reaction in the sentinel rabbit and a slight to moderate sting reaction in the second and third animal. Only minimal effects were observed after instillation of X14DesB30 to eyes of all three animals e.g. slight swelling of conjunctiva and production of ocular discharge. The sentinel rabbit showed a slight injection of the iris and conjunctiva and slight conjunctival swelling, that all had completely resolved by 48 hours. No corneal or iridal changes were apparent for the second and third rabbit. All reactions had overtly resolved within 48 hours of instillation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Insulin DesB30 is, based on read-across from in vivo data on MI3 and X13DesB30, not considered to be a skin or eye irritant. Thus, no CLP-classification should apply for skin and eye irritation of the substance.