Reaction products of ricinoleic acid and linoleic acid and oleic acid with sodium hydroxide and tin (II) chloride

Endpoint summary

Description of key information

ZINN(II)-RICINOLEAT does not require a classification for skin and/or eye irritation according to CLP, EU GHS (Regulation (EC) No 1272/2008) based on results from a primary dermal irritation study (OECD guideline 404) and an eye irritation study according to OECD guideline 405.

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

Study period:

2000-07-05 to 2000-07-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Species : Albino Rabbit, New Zealand White, (SPF-Quality). Recognised by international guidelines as the recommended test system (e.g. EC, OECD)
Source : Charles River Deutschland, Kisslegg, Germany.
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 less than 3.5 kg.
Identification : Earmark.
Conditions : A controlled environment was maintained in the room with optimal conditions considered as being approximately 15 air changes per hour, a temperature of 21°C, a relative humidity of 50% and 12 hours artificial fluorescent light and 12 hours dark per day. Deviations from these optimal conditions were noted, but were considered not to have affected study integrity.
Accommodation : Individually housed in labelled cages with perforated floors (Scanbur, Denmark) and equipped with an automatic drinking system (ITL, Bergen, The Netherlands). Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.
Diet : Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany) approx. 100 g. per day. Certificates of analysis were examined and retained in the NOTOX archives. In addition, hay (BMI, Helmond, the Netherlands) was provided once a week.
Water : Free access to tap-water. Certificates of quarterly analysis were examined and retained in the NOTOX archives.

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

other: adjacent area of untreated skin

Amount / concentration applied:

Each animal was treated by dermal application of 0.5 ml of the test substance.

Duration of treatment / exposure:

The test substance was applied to the skin of one flank, using a metalline patch# of 2x3 cm. The patch was mounted on Micropore tape*, which was wrapped around the abdomen and secured with Coban elastic bandage*. Four hours after the application, the dressing was removed and the skin cleaned of residual test substance using water and ethanol.
* Supplier: Lohmann GmbH, Neuwied, Germany.
* Supplier: 3M, St. Paul, Minnesota, U.S.A.

Observation period:

The skin reactions were assessed at approximately 1 , 24, 48 and 72 hours and 7 days after the removal of the dressings and test substance.

Number of animals:

3

Details on study design:

Approximately 24 hours before treatment, the dorsal fur was clipped with electric clippers, exposing an area of approximately 150 square centimeters (1 Ox15 cm). Whenever considered necessary the treated skin areas were re-clipped at least 3 hours before the observations, to facilitate scoring.
A health inspection was performed prior to the commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the skin to be treated, which was intact and free from abnormalities.
Observations:
Mortality/viability : Twice daily.
Toxicity : At least once daily.
Body Weight : Day of treatment (prior to application).
Irritation : The skin reactions were assessed at approximately 1 , 24, 48 and 72 hours and 7 days after the removal of the dressings and test substance. The irritation scores and a description of all other (local) effects were recorded. Adjacent areas of the untreated skin of each animal served as controls.
No histopathology was performed, since the skin reactions were not masked by test substance staining.

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3

Time point:

other: mean 24, 48, 72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Scaliness was noted in all animals at termination (after 7 days), which was considered to be a reversible skin reaction.

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: mean 24, 48, 72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: mean 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: mean 24, 48, 72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Exposure resulted in well-defined erythema and slight oedema in the treated skin-areas, which had been resolved within 7 days after exposure in all animals. Scaliness was noted in all animals at termination
(after 7 days). The scaliness, seen at termination, was considered to be a reversible skin reaction.

Other effects:

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

Individual irritation scores:

Animal number:	1 hour	24 hours	48 hours	72 hours	7 days	Average 24 – 72 hours
Erythema:
1	2	2	2	2	0	2
2	2	2	2	2	0	2
3	2	2	2	2	0	2
Edema:
1	2	1	0	0	0	0.33
2	2	1	1	1	0	1
3	2	1	1	0	0	0.67

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: other: CLP, EU GHS (Regulation (EC) No 1272/2008)

Conclusions:

Interpretation is based on erythema/eschar and edema calculated as mean scores for each animal following grading at 24, 48 and 72 hours after removal of the test material, in consideration of reversibility.The scaliness, seen at termination, was considered to be a reversible skin reaction.
According to CLP, EU GHS (Regulation (EC) No 1272/2008) no classification is required for ZINN(II)-RICINOLEAT

Executive summary:

Primary skin irritation/corrosion study with ZINN(II)-RICINOLEAT in the rabbit (4-hour semiocclusive application).

The study was carried out based on the guidelines described in: EC Commission Directive 92/69/EEC, 8.4, "Acute Toxicity - Skin irritation" and OECD No 404, "Acute Dermal Irritation/Corrosion" . Three rabbits were exposed to 0.5 ml of ZINN(II)-RICINOLEAT, applied onto clipped skin for 4 hours using a semi-occlusive dressing. Observations were made 1, 24, 48 and 72 hours and 7 days after exposure. Exposure to ZINN(II)-RICINOLEAT resulted in well-defined erythema and slight oedema in the treated skin-areas of the three rabbits. The skin irritation had resolved within 7 days after exposure in all animals. Scaliness was noted in all animals at termination (after 7 days). Remnants of the test substance were present on the skin of all animals on day 1. The scaliness, seen at termination, was considered to be a reversible skin reaction.

 

Interpretation of study results is based on mean scores for each animal following grading at 24, 48 and 72 hours for erythema and oedema: 

 

·      according to CLP, EU GHS (Regulation (EC) No 1272/2008): not irritating

(mean score erythema or edema 24, 48 , 72 hours of min. 2 of 3 is ≥ 2.3 - ≤ 4.0)

 

·      according to Directive 67/548/EEC: irritating R38

(mean score erythema or edema 24, 48, 72 hours of min. 2 of 3 is ≥ 2) 

The reported scaliness at study termination after 7 days was considered to be a reversible skin reaction.

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

Study period:

2000-07-10 to 2000-07-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline 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

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Species : Albino Rabbit, New Zealand White, (SPF-Quality). Recognised by international guidelines as the recommended test system (e.g. EC, OECD)
Source : Charles River Deutschland, Kisslegg, Germany.
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 less than 3.5 kg.
Identification : Earmark.
Conditions : A controlled environment was maintained in the room with optimal conditions considered as being approximately 15 air changes per hour, a temperature of 21 DC, a relative humidity of 50% and 12 hours artificial fluorescent light and 12 hours dark per day. Deviations from these optimal conditions were noted, but were considered not to have affected study integrity.
Accommodation : Individually housed in labelled cages with perforated floors (Scanbur, Denmark) and equipped with an automatic drinking system (ITL, Bergen, The Netherlands). Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.
Diet : Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany) approx. 100 g. per day. Certificates of analysis were examined and retained in the NOTOX archives. In addition, hay (BMI, Helmond, the Netherlands) was provided once a week.
Water : Free access to tap-water. Certificates of quarterly analysis were examined and retained in the NOTOX archives.

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated eye served as control

Amount / concentration applied:

Each animal was treated by instillation of 0.1 ml of the test substance as tile maximum required amount, in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball.

Duration of treatment / exposure:

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.

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48 and 72 hours after instillation of the test substance

Number of animals or in vitro replicates:

3

Details on study design:

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.
Mortality/Viability : Twice daily.
Toxicity : At least once daily.
Body Weight : Day of treatment (prior to instillation).
Irritation : The eyes of each animal were examined approximately 1,24,48 and 72 hours after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded.

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3

Time point:

other: mean 24, 48, 72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

other: mean 24, 48, 72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #3

Time point:

other: mean 24, 48, 72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: mean 24, 48, 72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal: #2, #3

Time point:

other: mean 24, 48, 72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: mean 24, 48, 72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritant / corrosive response data:

Exposure resulted in irritation of the conjunctivae , which was seen as redness, chemosis and discharge. The irritation had completely resolved within 48 hours in one animal and within 72 hours in the other two animals.
No iridial irritation or corneal opacity were observed, and treatment of the eyes with 2 % fluorescein, 24 hours after test substance instillation
revealed no corneal epithelial damage in any of the animals.

Other effects:

Colouration: No staining of peri-ocular tissues by the test substance was observed.
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Irritant/corrosive response data for each animal at each observation time up to removal of each animal from the test

Score at time point/ Reversibility	Cornea	Iris	Conjunctivae	Chemosis
	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4
60 min	0/0/0	0/0/0	1/1/1	0/1/0
24 h	0/0/0	0/0/0	2/1/2	1/0/0
48 h	0/0/0	0/0/0	1/0/1	0/0/0
72 h	0/0/0	0/0/0	0/0/0	0/0/0
Average 24h, 48h, 72h	0/0/0	0/0/0	1/0.33/1	0.33/0/0
Reversibility*)			c.	c.
Average time (unit) for reversion			72 h	72 h

*) Reversibility: c. = completely reversible; n.c.= not completely reversible; n. = not reversible

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: CLP, EU GHS (Regulation (EC) No 1272/2008)

Conclusions:

In this study the test substance is not irritating to the eye. Interpretation is based on effects on the cornea, iris and conjunctiva calculated as mean scores following grading at 24, 48 and 72 hours after instillation of the test material, in consideration of reversibility.
According to CLP, EU GHS (Regulation (EC) No 1272/2008) no classification is required for ZINN(II)-RICINOLEAT

Executive summary:

Acute eye irritation/corrosion study with ZINN(II)-RICINOLEAT in the rabbit. The study was carried out based on the guidelines described in: EC Commission Directive 92/69/EEC, B.5, "Acute Toxicity - Eye irritation" and OECD No 405, "Acute Eye Irritation/Corrosion" .

Single samples of 0.1 ml of ZINN(II)-RICINOLEAT were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours after instillation. Instillation of the test substance resulted in irritation of the conjunctivae, which was seen as redness, chemosis and discharge. The irritation had completely resolved within 48 hours in one animal and within 72 hours in the other two animals.

No iridial irritation or corneal opacity were observed, and treatment of the eyes with 2 % fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage in any of the animals.

Interpretation of study results is based on mean scores for each animal following grading at 24, 48 and 72 hours for cornea, iris, conjuctiva and chemosis: 

No classification for eye irritation is required for ZINN(II)-RICINOLEAT according to CLP, EU GHS (Regulation (EC) No 1272/2008) and Directive 67/548/EEC.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

In a primary dermal irritation study according to OECD guideline 404, three New Zealand White rabbits were dermally exposed to 0.5 g of ZINN(II)-RICINOLEAT, undiluted as delivered by the sponsor. The test substance was applied for 4 hours to intact shaved body surface areas under semi-occlusive dressing. Animals then were observed for 7 days. Irritation was scored by the method described in OECD Guideline 404.

Exposure resulted in well-defined erythema (max. mean score 2)and slight oedema (max. mean score 1) in the treated skin-areas of the three rabbits. The skin irritation had resolved within 7 days after exposure in all animals. Scaliness was noted in all animals at termination (after 7 days), which was considered to be a reversible skin reaction.

 

Eye irritation:

In a primary eye irritation study according to OECD Guideline 405, 0.1 mL of ZINN(II)-RICINOLEAT, was instilled undiluted into the conjunctival sac of one eye of three New Zealand White rabbits. The untreated eye served as control. The eyes were not washed. Animals were then observed for 7 days.

Instillation of the test substance resulted in irritation of the conjunctivae, which was seen as redness, chemosis and discharge. The irritation had completely resolved within at least 72 hours in all animals. No iridial irritation or corneal opacity were observed, and treatment of the eyes with 2 % fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage in any of the animals.

Justification for selection of skin irritation / corrosion endpoint:
Data from a GLP compliant guideline study with reliability 1.

Justification for selection of eye irritation endpoint:
Data from a GLP compliant guideline study with reliability 1.

Justification for classification or non-classification

Interpretation is based on study results from a primary dermal irritation study (OECD guideline 404) and an eye irritation study according to OECD guideline 405.

Skin irritation:

Evaluation is based on mean scores for each animal following grading at 24, 48 and 72 hours for erythema and oedema. The max. mean score was 2 (for erythema) in all three animals.

The following classification is required for ZINN(II)-RICINOLEAT:

·      according to CLP, EU GHS (Regulation (EC) No 1272/2008): not irritating

(mean score erythema or edema 24, 48 , 72 hours of min. 2 of 3 is ≥ 2.3 - ≤ 4.0)

·      according to Directive 67/548/EEC: irritating R38, (may cause irritation)

(mean score erythema or edema 24, 48, 72 hours of min. 2 of 3 is ≥ 2) 

The reported scaliness at study termination after 7 days was considered to be a reversible skin reaction.

 

Eye irritation:

Interpretation of study results is based on mean scores for each animal following grading at 24, 48 and 72 hours for cornea, iris, conjuctiva and chemosis: 

No classification for eye irritation is required for ZINN(II)-RICINOLEAT according to CLP, EU GHS (Regulation (EC) No 1272/2008) and Directive 67/548/EEC.