Bis(6-methylalkyl-5-en-2-yl)dihydro-heteropolycycle

Endpoint summary

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-Sept.-04 until 2007-Nov.-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Version / remarks:

1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF guidelines (2000) including the most recent partial revisions

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST SYSTEM:

Species: Albino rabbit, New Zealand White, (SPF-Quality). Recognised by international guidelines as the recommended test system (e.g. EC, OECD)
Source: Harlan, Horst, The Netherlands.

Number of animals: 3 Males.

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.

Health inspection: 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.

ANIMAL HUSBANDRY

Conditions: Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 21.0 ± 3.0 °C (actual range: 20.9 - 24.0 °C), a relative humidity of 30-70% (actual range: 39 - 67%) and 12 hours artificial fluorescent light and 12 hours darkness 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: Pelleted diet for rabbits (K-H from SSNIFF® Spezialdiäten GmbH, Soest, Germany) approximately 100 grams per day. Hay (TecniLab-BMI BV, Someren, The Netherlands) was provided at least three times a week.

Water: Free access to tap water.

Results of analysis for each batch of diet (nutrients and contaminants), hay and water were assessed and did not reveal any findings that were considered to have affected the study integrity. All certificates and results of analysis are retained in the NOTOX archives

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

Purity: According to substance identity report
0,5 mL of the undiluted test substance was applied.

Duration of treatment / exposure:

4 hour(s)

Observation period:

1, 24, 48 and 72 hours and 7 days after the removal of the dressings and test substance.

Number of animals:

3 males

Details on study design:

Study design: The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 2 weeks later, after considering the degree of skin irritation observed in the first animal.

Treatment: Approximately 24 hours before treatment, the dorsal fur was clipped with electric clippers, exposing an area of approximately 150 square centimeters (10x15 cm). To facilitate scoring, treated skin areas were re-clipped at least 3 hours before the observations.
Each animal was treated by dermal application of 0.5 mL of the test substance. 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 tap water.
#. Suppliers: Lohmann GmbH, Neuwied, Germany (Metalline) and 3M, St. Paul, Minnesota, U.S.A. (Micropore and Coban).
After the final observation, the animals were sacrificed by intra-venous injection of pentobarbital Euthesate® (Ceva Sante Animale BV, Naaldwijk, The Netherlands).

Observations:

Mortality/Viability: Twice daily.

Toxicity: At least once daily.

Body Weight: Day of treatment (prior to application) and at termination.

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.

Irritation parameter:

erythema score

Basis:

animal: 518 (Sentinel)

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: average

Irritation parameter:

erythema score

Basis:

animal: 549

Time point:

24/48/72 h

Score:

1.3

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: average

Irritation parameter:

erythema score

Basis:

animal: 552

Time point:

24/48/72 h

Score:

1.7

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: average

Irritation parameter:

edema score

Basis:

animal: 518 (Sentinel)

Time point:

24/48/72 h

Score:

1.7

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: average

Irritation parameter:

edema score

Basis:

animal: 549

Time point:

24/48/72 h

Score:

0.3

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: average

Irritation parameter:

edema score

Basis:

animal: 552

Time point:

24/48/72 h

Score:

0.7

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: average

Irritant / corrosive response data:

Irritation: Four hours exposure to 0.5 mL of Sa03 resulted in moderate to severe (focal) erythema and slight oedema in the treated skin-areas of the three rabbits. Scaliness was present in all animals 72 hours. The skin irritation had completely resolved within 7 days after exposure in all animals.

Corrosion: There was no evidence of a corrosive effect on the skin.

Other effects:

Colouration/Remnants: Sticky and dry remnants of the test substance were present on the skin on Day 1.
Toxicity/Mortality: No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occured.

Interpretation of results:

GHS criteria not met

Conclusions:

According to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2003), the test item should be classified as: mild skin irritant (Category 3).
According to the CLP Regulation (EC) 1272/2008, Sa03 does not have to be classified and has no obligatory labelling requirement for skin irritation.

Executive summary:

Primary skin irritation/corrosion study with Sa03 in the rabbit (4-hour semi-occlusive application).

 

The study was carried out based on the guidelines described in:

OECD No.404, "Acute Dermal Irritation/Corrosion" (2002)

EC, Council Directive 67/548/EEC, An.V, B.4 (2004), "Acute Toxicity: Dermal Irritation/ Corrosion"

USEPA, OPPTS 870.2500 (1998), Acute Dermal Irritation 

JMAFF Guidelines (2000) including the most recent partial revisions.

  

Three rabbits were exposed to 0.5 mL of Sa03 by application onto clipped skin for 4 hours using a semi-occlusive dressing. Skin reactions were assessed 1, 24, 48 and 72 hours and 7 days after exposure.

  

Exposure to Sa03 resulted in moderate to severe (focal) erythema and slight oedema in the treated skin-areas of the three rabbits. Scaliness was present in all animals 72 hours. The skin irritation had completely resolved within 7 days after exposure in all animals.

 

Sticky and dry remnants of the test substance were present on the skin on Day 1.

  

Based on these results:

-       according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2003), Sa03 should be classified as : mild skin irritant (Category 3).

 

- according to the CLP Regulation (EC) 1272/2008, Sa03 does not have to be classified and has no obligatory labelling requirement for 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 vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2006

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study meets generally accepted scientific principles

Qualifier:

no guideline available

Principles of method if other than guideline:

ln vitro assay: HET-CAM [HCI], Reaction-Time Method;
The aim of the test is the evaluation of the irritation potential of the test substance on the chorionallantoic membrane (CAM) of fertilized chicken eggs, in order to estimate the local irritation property of the substances after contact with mucous membranes or eyes.

GLP compliance:

yes

Remarks:

The study was conducted in compliance with the regulations of "Good_ Laboratory- Practice" (GLP).

Species:

other: HET-CAM

Strain:

not specified

Details on test animals or tissues and environmental conditions:

The irritation potential of the test substance was tested on the chorionallantoic membrane (CAM) of fertilized chicken eggs, in order to estimate the local irritation property of the substances after contact with mucous membranes or eyes.

Vehicle:

other: undiluted and with vehicle olive oil

Controls:

not required

Amount / concentration applied:

In this study the test substance Sa03 was tested undiluted and as a 50% dilution in olive oil by the reaction-time method.
The test substance was added in approx. 300 µl to six eggs with at least well developed blood vessels. The amount of the added test substance covered approximately 25% CAM.

Duration of treatment / exposure:

30 sec.

Observation period (in vivo):

5 minutes

Number of animals or in vitro replicates:

0

Details on study design:

Reaction-Time Method
Transparent test substances were tested by the reaction-time method. The reaction-time period and reaction intensity of effects on the CAM like haemorrhage, lysis of the vessels and protein coagulation (intravascular and/or extravascular) were visually evaluated.
In order to compare the results of in vitro with in vivo experiments and to minimize effects due to biological variation of the charges of the eggs, an irritation index of the reference substance "Texapon ASV" was calculated [5% active substance, pH approx. 7]. This concentration was classified as being moderately irritating to rabbit's eyes.
The required dilutions (percent of weight) of the reference substance were made in bidistilled water and the dilutions of the test substance were made in olive oil.

Irritation parameter:

other: Reaction-Time Method (Q)

Value:

0

Positive controls validity:

valid

Remarks on result:

other: The undiluted test substance induced no reaction in the reaction-time method till five minutes.

Remarks:

In a pre-experiment the test substance was tested 50% diluted in olive oil, also no reaction was observed. Under these test conditions test substance Sa03 was predicted to be slightly irritating. Based on our experience in comparing in vitro and in vivo results we can suppose that the test substance induce the same irritations after the contact with the mucous membrane, particularly with the eye (Draize-Test).

Test substance Sa03

Reaction Time Method Q: 0.00

Evaluation: slighly irritating

 

Reference Substance, 5% AS

Reaction Time Method Q: 1.00, SD +/-0.05

Evaluation: moderately irritating

Interpretation of results:

slightly irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: internal SOP Henkel

Conclusions:

The undiluted test substance induced no reaction in the reaction-time method till five minutes.
In a pre-experiment the test substance was tested 50% diluted in olive oil, also no reactions were observed.
Under these test conditions test substance Sa03 was predicted to be slightly irritating.
Based on our experience in comparing in vitro and in vivo results we can suppose that the test substance induce the same irritations after the contact with the mucous membrane, particularly with the eye (Draize-Test).

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available

Reference 3

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Sep - Nov 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2004

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

1998

Qualifier:

according to guideline

Guideline:

other: JMAFF guidelines (2000); including the most recent partial revisions

Version / remarks:

2000

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST SYSTEM

Species: Albino rabbit, New Zealand White, (SPF-Quality). Recognized by international guidelines as the recommended test system (e.g. EC, OECD)
Source: Harlan, Horst, The Netherlands.

Number of animals: 3 Males.

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.

Health inspection: 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.

ANIMAL HUSBANDRY

Conditions: Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 21.0 ± 3.0°C (actual range: 21.4 – 24.0°C), a relative humidity of 30-70% (actual range: 44 - 68%) and 12 hours artificial fluorescent light and 12 hours darkness per day.

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

Diet: Pelleted diet for rabbits (K-H from SSNIFF® Spezialdiäten GmbH, Soest, Germany) approximately 100 grams per day. Hay (TecniLab-BMI BV, Someren, The Netherlands) was provided at least three times a week.

Water: Free access to tap water.

Results of analysis for each batch of diet (nutrients and contaminants), hay and water were assessed and did not reveal any findings that were considered to have affected the study integrity. All certificates and results of analysis are retained in the NOTOX archives.

Vehicle:

unchanged (no vehicle)

Controls:

other: second eye served as the reference control

Amount / concentration applied:

Purity: According to substance idenity report
0.1 mL of undiluted test substance was applied

Duration of treatment / exposure:

24 hour(s)

Observation period (in vivo):

1, 24, 48 and 72 hours

Number of animals or in vitro replicates:

3 males

Details on study design:

Study design: The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner one week later, after considering the degree of eye irritation observed in the first animal.

Treatment: Each animal was treated by instillation of 0.1 ml of the test substance in the conjunctival 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 (Merck, Darmstadt, Germany) in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. Any bright green stained area, indicating epithelial damage,
was estimated as a percentage of the total corneal area. Since no residual test substance was seen, the treated eyes were not rinsed with tepid tap water.
After the final observation, the animals were sacrificed by intra-venous injection of pentobarbital Euthesate® (Ceva Sante Animale BV, Naaldwijk, The Netherlands).

Observations:

Mortality/Viability: Twice daily.

Toxicity: At least once daily.

Body Weight: Day of treatment (prior to instillation) and at termination.

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: 530 (Sentinel)

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no irritation

Remarks on result:

other: average

Irritation parameter:

cornea opacity score

Basis:

animal: 550

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no irritation

Remarks on result:

other: average

Irritation parameter:

cornea opacity score

Basis:

animal: 551

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no irritation

Remarks on result:

other: average

Irritation parameter:

iris score

Basis:

animal: 530 (Sentinel)

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no irritation

Remarks on result:

other: average

Irritation parameter:

iris score

Basis:

animal: 550

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no irritation

Remarks on result:

other: average

Irritation parameter:

iris score

Basis:

animal: 551

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no irritation

Remarks on result:

other: average

Irritation parameter:

conjunctivae score

Basis:

animal: 530 (Sentinel)

Time point:

24/48/72 h

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: average; score 1 after 1 hour

Irritation parameter:

conjunctivae score

Basis:

animal: 550

Time point:

24/48/72 h

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: average; score 1 after 1 hour

Irritation parameter:

conjunctivae score

Basis:

animal: 551

Time point:

24/48/72 h

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: average; score 1 after 1 hour

Irritation parameter:

chemosis score

Basis:

animal: 530 (Sentinel)

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no irritation

Remarks on result:

other: average

Irritation parameter:

chemosis score

Basis:

animal: 550

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no irritation

Remarks on result:

other: average

Irritation parameter:

chemosis score

Basis:

animal: 551

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no irritation

Remarks on result:

other: average

Irritation parameter:

other: conjunctivae discharge

Basis:

animal: 530 (Sentinel)

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no discharge

Remarks on result:

other: average

Irritation parameter:

other: conjunctivae discharge

Basis:

animal: 550

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, no discharge

Remarks on result:

other: average

Irritation parameter:

other: conjunctivae discharge

Basis:

animal: 551

Time point:

24/48/72 h

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: average; score 1 after 1 hour

Irritant / corrosive response data:

Irritation:
Instillation of 0.1 mL of Sa03 into one eye of each of three rabbits resulted in irritation of
the conjunctivae, which consisted of redness and discharge. The irritation had completely resolved within 24 hours. No iridial irritation or corneal opacity was observed, and treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage.

Corrosion:

There was no evidence of ocular corrosion.

Other effects:

Colouration/Remnants: No staining of (peri) ocular tissues by the test substance was observed and no test substance remnants were seen.

Toxicity/Mortality: No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occured.

Interpretation of results:

GHS criteria not met

Conclusions:

Based on these results Sa03 does not have to be classified and has no obligatory labelling requirement for eye irritation according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2004) and to the CLP Regulation (EC) 1272/2008.

Executive summary:

Acute eye irritation/corrosion study with Sa03 in the rabbit.

 

The study was carried out based on the guidelines described in:

OECD No.405 (2002) "Acute Eye Irritation / Corrosion"

EC, Council Directive 67/548/EEC, An. V,B.5, (2004) "Acute Toxicity: Eye Irritation / Corrosion"

EPA, OPPTS 870.2400 (1998): "Acute Eye Irritation"

JMAFF guidelines (2000); including the most recent partial revisions.

  

Single samples of 0.1 ml of Sa03 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 consisted of redness and discharge. The irritation had completely resolved within 24 hours.

  

Based on these results Sa03 does not have to be classified and has no obligatory labelling requirement for eye irritation according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2004) and to the CLP Regulation (EC) 1272/2008.

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

The test substance did not produce skin irritation and eye irritation. The substance does not need to be classified for irritation according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.