4-methyl-3-decen-5-ol

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

key study

Study period:

21 June 2010 to 13 August 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

A valid study is available for the analogue substance (2E)−3,7−dimethylocta−2,6−dien−1−ol. It is a GLP compliant study conducted in compliance with agreed protocols, with no or minor deviations from standard testing guidelines The read-across is considered to be suitable based on the structural and “mechanistic action” similarities between the target substance (4-methyl-3-decen-5-ol) and source substance ((2E)−3,7−dimethylocta−2,6−dien−1−ol) and their similar physico-chemical properties.

Justification for type of information:

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that the target substance and 1 source substances have the same expected mode of action and similar physicochemical properties relevant for the read-across endpoints.
The justification of the proposed read-across to geraniol is discussed in the attached RAAF document.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The target substance (undecavertol) is a mono-constituent substance (EC 279-815-0, CAS 81782-77-6). The typical concentration of the single constituent is 98.0%.
The source substance geraniol (EC 203-377-1, CAS 106-24-1) is a mono-constituent substance. The typical concentration of the mono-constituents is 97.0%.
The target substance and the source substance do not contain any impurities present at ≥ 1%. The purity of the test items within the respective REACH registration dossiers for undecavertol and for geraniol indicates purity > 97.0% with no impurities > 1%.

3. ANALOGUE APPROACH JUSTIFICATION
The structures of the target and source substance are provided in Table 1 (RAAF document). The target substance and the source substance have been characterised in this table using the categories and databases present in the OECD [Q]SAR Toolbox. From the profiling provided (Table 1 - (RAAF document)), it can be seen that the 2 substances share structural similarities and also mechanistic actions which are both general and endpoint specific. This supports the hypothesis that the target and source substances have similar properties as a result of structural similarity and the same expected mode of action.
The OECD toolbox predicts all substances to be of low toxicity according to Cramer classes and both substances show no alerts according to DART Scheme v1.0.
Undecavertol and geraniol are structurally similar substances. The primary route of metabolism for undecavertol is aliphatic c-oxidation followed by either o-glucoronidation or beta oxidation. Geraniol is metabolised via epxoidation foolwed by aliphatic c-oxidation. This is supported by the most probable route of metabolism prediction of TIMES v.2.27.17 (rat in vivo model) as illustrated in the RAAF document..

4. DATA MATRIX
Please see the RAAF document.

Cross-reference

Reference

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

key study

Study period:

21 June 2010 to 13 August 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

A valid study is available for the analogue substance (2E)−3,7−dimethylocta−2,6−dien−1−ol. It is a GLP compliant study conducted in compliance with agreed protocols, with no or minor deviations from standard testing guidelines The read-across is considered to be suitable based on the structural and “mechanistic action” similarities between the target substance (4-methyl-3-decen-5-ol) and source substance ((2E)−3,7−dimethylocta−2,6−dien−1−ol) and their similar physico-chemical properties.

Justification for type of information:

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that the target substance and 1 source substances have the same expected mode of action and similar physicochemical properties relevant for the read-across endpoints.
The justification of the proposed read-across to geraniol is discussed in the attached RAAF document.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The target substance (undecavertol) is a mono-constituent substance (EC 279-815-0, CAS 81782-77-6). The typical concentration of the single constituent is 98.0%.
The source substance geraniol (EC 203-377-1, CAS 106-24-1) is a mono-constituent substance. The typical concentration of the mono-constituents is 97.0%.
The target substance and the source substance do not contain any impurities present at ≥ 1%. The purity of the test items within the respective REACH registration dossiers for undecavertol and for geraniol indicates purity > 97.0% with no impurities > 1%.

3. ANALOGUE APPROACH JUSTIFICATION
The structures of the target and source substance are provided in Table 1 (RAAF document). The target substance and the source substance have been characterised in this table using the categories and databases present in the OECD [Q]SAR Toolbox. From the profiling provided (Table 1 - (RAAF document)), it can be seen that the 2 substances share structural similarities and also mechanistic actions which are both general and endpoint specific. This supports the hypothesis that the target and source substances have similar properties as a result of structural similarity and the same expected mode of action.
The OECD toolbox predicts all substances to be of low toxicity according to Cramer classes and both substances show no alerts according to DART Scheme v1.0.
Undecavertol and geraniol are structurally similar substances. The primary route of metabolism for undecavertol is aliphatic c-oxidation followed by either o-glucoronidation or beta oxidation. Geraniol is metabolised via epxoidation foolwed by aliphatic c-oxidation. This is supported by the most probable route of metabolism prediction of TIMES v.2.27.17 (rat in vivo model) as illustrated in the RAAF document..

4. DATA MATRIX
Please see the RAAF document.

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3550 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

other: Crl:WI(Han)

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Charles River Laboratories, Research Models and Services, Germany GmbH- Age at study initiation: approximately 11 - 12 weeks (at start of administration)- Weight at study initiation: 286.8 - 322.3 g (males); 189.8 - 219.5 g (females)- Housing: Individually in Makrolon type M III cages (Becker & Co., Castrop-Rauxel, Germany) except during mating when one male and one female were housed together in Makrolon type M III cages. Pregnant females and their litters were housed together until the end of lactation. Pregnant females were provided with nesting material (cellulose wadding) towards the end of gestation. - Use of restrainers for preventing ingestion (if dermal): no- Diet:ground KLiba maintenance diet mouse/rat "GLP" meal (Provimi Kliba SA, Kaiseraugst, Switzerland) ad libitum- Water: ad libitum- Acclimation period: 7 daysENVIRONMENTAL CONDITIONS- Temperature (°C): 20 - 24 °C- Humidity (%): 30 - 70 %- Air changes (per hr): 15 air changes per hour- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light

Route of administration:

dermal

Vehicle:

corn oil

Details on exposure:

TEST SITE- Area of exposure: Dorsal- Type of wrap if used: A semiocclusive dressing (4 layers of absorbent gauze and stretch bandage)- Time intervals for shavings or clipplings: The dorsal skin was clipped at least once a week depending on hair growth.REMOVAL OF TEST SUBSTANCE- Washing: After removal of the dressing, the application area was washed with lukewarm water.- Time after start of exposure: Performed dailyTEST MATERIAL- Amount(s) applied (volume or weight with unit): 4 mL/kg. The calculation of the volume administered was generally based on the most recent individual body weights.- Constant volume or concentration used: noVEHICLE- Amount(s) applied (volume or weight with unit): 4 mL/kgUSE OF RESTRAINERS FOR PREVENTING INGESTION: no

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analytical verification of test concentrations was performed on samples of all concentrations at the start and towards the end of the adminstration period. Concentration analysis was performed by HPLC with UV detection. The HPLC conditions were as follows:- Column: Gemini C18 5 µ 110A, 150 x 3 mm- Eluent: 50 % acetonitrile + 0.5 M sulphuric acid (5 mL/L); 50 % highly deionised water + 0.5 M sulphuric acid (5 mL/L)- Flow rate: 0.6 mL/min- Injection volume: 2, 5 µL- Column temperature: ambient- Detection: 205 nm- Limit of quantification: 12.8 mg/L

Details on mating procedure:

- M/F ratio per cage: one male and one female of the same treatment group - Length of cohabitation: animals were mated overnight (from 16:00 to 07:00 - 09:00 the following morning)- Further matings after two unsuccessful attempts: yes, animals were mated for a maximum of two weeks.- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

The duration of treatment covered the premating period of 2 weeks and mating period (maximum of 2 weeks) in both sexes, approximately 1 week post-mating in males, and the entire gestation period until gestation day 19 in females (although the females were not treated during the end of gestation and during lactation).

Frequency of treatment:

Daily (6 hours exposure per day, 7 days per week)

Duration of test:

Up to gestation day 19 for parental animals (with a two week pre-mating period and a two week (maximum) mating period), and postnatal day 4 for offspring

No. of animals per sex per dose:

10 females per dose level

Control animals:

yes

Details on study design:

- Dose selection rationale: In a previous study rats received dermal applications of test material at the dose levels 0, 300, 500, 750 and 1000 mg/kg bw/day for 14 days. Corn oil served as vehicle. In this study the test material caused scales and erythema at dose levels of 500 mg/kg bw/day and above but no findings at a dose level of 300 mg/kg bw/day. Therefore 450 mg/kg bw/day was selected as the high dose and 150 and 50 mg/kg bw/day were selected as intermediate and low doses, respectively.- Rationale for animal assignment: Animals were distributed according to weight among the individual test groups, separated by sex. The weight variation of the animals used did not exceed 20 % of the mean weight of each sex.

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes - Time schedule: mortality was checked twice daily and clinical observations were performed at least once dailyBODY WEIGHT: Yes - Time schedule for examinations: Weekly with the following exceptionsFemales were weighed on the day of positive evidence of sperm (GD 0), and on GD 7, 14 and 20 during mating.Females showing no positive sign of sperm in the vaginal smear were weighed once a week during mating as were malesFemales with litter were weighed on the day of parturition (PND 0) and on PND 4.Females without litter were weighed once a weekFemaels between PND 4 and sacrifice were weighed once a week.FOOD CONSUMPTION: Yes- Time schedule for examinations: weekly (food consumption was not determined in the females without positive evidence of sperm during mating and gestation periods and in the females without litter during lactation period) Food consumption was not determined during the mating periodFood consumption of the females with evidence of sperm was determined for GD 0-7, 7-14 and 14-20.Food consumption for females which gave birth to a litter was determined for PND 1-4.WATER CONSUMPTION: NoOTHER: The parturition and lactation behaviour of the dams was generally evaluated once daily. The day of parturition was considered to be the 24 hour period from about 15:00 of one day until about 15:00 of the following day.SACRIFICE: Parental animals were sacrificed by decapitation under isoflurane anesthesia. The exanguinated animals were necropsied and assessed by gross pathology.- Sacrifice on gestation day 19Animals which died intercurrently or were sacrificed in a moribund state were necropsied as soon as possible after their death and assessed by gross pathology.ORGAN WEIGHTSThe weights of the following were determined: carcass and ovariesORGAN/TISSUE FIXATIONThe following were fixed in 4 % neutral buffered formaldehyde solution or in modified Davidson's solution: all gross lesions, adrenal glands, ovaries, pituitary gland, coagulation glands, skin (treated), skin (untreated), uterus, oviducts, vagina.The ovaries of animals that died or were sacrificed intercurrently were fixed in 4 % buffered formaldehyde solution.HISTOPATHOLOGYAfter organs were fixed, histotechnical processing and examination by light microscopy were performed on gross lesions and treated skin of animals of all test groups and on the ovaries and untreated skin in the control and high dose group animals.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: YesExaminations included:- Number of implantations: Yes

Fetal examinations:

PARAMETERS EXAMINEDThe following parameters were examined in F1 offspring: number and sex of pups, stillbirths, live births, gross-morphological changes. On post-natal day 1 live pups were weighed.On post-natal day 4, the pups were sacrificed under isoflurane anesthesia with CO2. After sacrifice, the pups were examined externally and eviscerated, and their organs were assessed macroscopically.Pups that died or were sacrificed in a moribund state were eviscerated and examined for possible defects and/or the cause of death.All pups without any notable findings were discarded after their macroscopic evaluation.

Statistics:

Dunnett’s-test (two-sided) for the hypothesis of equal means was performed on the following parameters: food consumption; body weight; body weight change; number of mating days; duration of gestation; number of pups delivered per litter; implantation sites and post implantation loss. Pair-wise comparison of each dose group with the control group was performed with Fisher’s Exact test on the following parameters: mating indices; fertility indices; gestation index; females with liveborn and stillborn pups; live birth index, stilibrin, dead, cannibalised, sacrificed and moribund pups; viability index and number of litters with affected pups at necropsy. Pairwise comparison of each dose group was analysed with the Wilcoxon-test on the parameters: portions of affected pups per litter with necropsy observations.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

See "Details on results (parental animals)" for information

Dermal irritation (if dermal study):

effects observed, treatment-related

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Other effects:

not examined

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

not specified

Dead fetuses:

no effects observed

Changes in pregnancy duration:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed

Changes in number of pregnant:

no effects observed

Other effects:

not specified

Details on maternal toxic effects:

Maternal toxic effects:no effects. Remark: Only local signs of irritation were reportedDetails on maternal toxic effects:CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)Female no. 127 (dosed at 150 mg/kg bw/day) was sacrificed moribund on study day 11 because of a severe thoracic injury. No test material-related mortalities occurred in any test group.For female animals, different dermal findings were seen on several days. In test group 3, 7 females showed slight and moderate erythema on several days of the study, starting on study day 3. In addition, some animals of test group 3 showed multifocal, focal and diffuse scales on treated skin starting on study day 5. Similar findings but less pronounced were observed in female animals of test group 2, i.e. focal red spots, slight erythema and focal scales on tre6aed skin on several days of the study. In test group 1, slight erythema as well as focal and diffuse scales on treated skin were observed in individual animals at different time points.Female no. 137 of test group 3, which did not deliver pups, showed a vaginal haemorrhage 27 days after mating.In female animal 127 of test group 2, a severe thoracal injury was observed during pre-mating. It was a self inflicted injury which was caused by the animal's attempt to get rid of the gauze. The injury became more severe by time and the animal had to be sacrificed in a moribund state on study week 1.BODY WEIGHTThe mean body weight and body weight change of animals showed no significant deviations to the control group in all test groups.FOOD CONSUMPTION No changes in food consumption of toxicological significance were seen for male and female animals of all test groups.ORGAN WEIGHTSNone of the absolute of relative weight parameters in test groups 1 - 3 showed relevant differences when compared to the control group and were considered to be within the normal range.GROSS PATHOLOGYA red-brown lesion was noted on the thorax of female no. 127 of test group 2 which was sacrificed moribund. All gross lesions observed in test animals occurred singularly. They were considered to be spontaneous lesions in origin and not related to treatment.HISTOPATHOLOGY The lesion noted on the thorax of female no. 127 correlated to an erosion on the skin. Lymphocytic infiltrates were observed in treated skin sections which were distributed band-like between epidermis and dermis. All other findings noted were single observations which were considered to be incidental and spontaneous in origin and without any relation to treatment.

Key result

Dose descriptor:

NOAEL

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Key result

Dose descriptor:

NOAEL

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Fetal body weight changes:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Visceral malformations:

no effects observed

Other effects:

not specified

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effectsDetails on embryotoxic / teratogenic effects:VIABILITY (OFFSPRING)No significant findings for Pups that died during lactation were observed. The viability index varied between 99 % for test groups 1 and 3 and 100 % for test groups 0 and 2.No test material-related changes were obtained.Male pup no. 8, delivered by dam no 118 of test group 1, was sacrificed moribund on post natal day 0 as it showed malformation of the skull, anophthalmia and cleft lip.The sex distribution and sex ratios of live F1 pups on the day of birth and post natal day 4 did not show biologically relevant differences between the control and test groups 1 to 3.CLINICAL SIGNS (OFFSPRING)Dam no 118 (test group 1) gave birth to male pup no 8 with a deformation of its snout. On first sight a cleft lip and anophthalmia were seen. The staining of the pups skull showed that several bones (basisphenoid, palatine, incisive, nasal - including cartilage and maxilla) were deformed and/or displaced. therefore, F1 pups did not show adverse clinical signs up to schedules sacrifice on post natal day 4.BODY WEIGHT (OFFSPRING)The mean body weight of female pups was increased by 10 % on post natal day 4 in test group 3. Pup body weight change in groups 1 to 3 were comparable to the concurrent control values.

Key result

Dose descriptor:

NOAEL

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no effects observed

Abnormalities:

not specified

Developmental effects observed:

not specified

Homogeneity and concentration control analyses

The test material was completely miscible with Corn Oil Ph. Eur, and thus, a solution. Therefore, the test material preparation was considered to be homogenous and no further homogeneity analysis was performed.

The concentration control analyses at the beginning and towards the end of the application period revealed values in the expected range of the target concentrations. the means of the nominal concentrations were in a range of 96.5 - 104.3 % of the nominal concentrations.

ANALOGUE APPROACH JUSTIFICATION:

- See attached “Justification for read-across” document for full details.

- In summary, important considerations for the use of read-across for reproductive/developmental toxicity screening are: i) 4-methyldec-3-en-5-ol (the target chemical) has similar physico-chemical properties as 4(2E)−3,7−dimethylocta−2,6−dien−1−ol (the source substance), ii) there are structural similarities between the two chemicals and iii) the OECD QSAR Toolbox assigns an identical toxicity profiles to both chemicals.

The information reported in this summary is included to demonstrate comparability between the source (4(2E)−3,7−dimethylocta−2,6−dien−1−ol) and target (4-methyldec-3-en-5-ol) substance.

Conclusions:

Under the conditions of the study the No Observed Adverse Effect Level (NOAEL) for maternal toxicity was determined to be 300 mg/kg bw/day. The NOAEL for developmental toxicity was determined to be 300 mg/kg bw/day.

Executive summary:

The reproductive and developmental toxicity of the test material was investigated in a GLP study which was conducted in accordance with standardised guidelines OECD 421 and EPA OPPTS 870.3550. During the study test material was administered via dermal administration to groups of 10 male and 10 female Wistar rats at dose levels of 0 (vehicle control; test group 0), 50 (test group 1), 150 (test group 2) and 450 mg/kg (test group 3) in order to observe the possible effects of the test material on the integrity and performance of the reproductive system in both sexes. Due to severe dermal findings, the dose level for test group 3 was decreased to 300 mg/kg bw/day from study day 10 onwards. Only signs of local dermal toxicity were observed for males and females at all dose levels. No changes in food consumption or body weight were seen at any dose level. fertility indices for male and female animals were not impaired by test material administration. Furthermore, there were no treatment-related or histological findings in ovaries, testes or epipdiymides associated with dermal administration of the test material. The local minimal inflammatory reactions in the skin of treated males (test groups 1 to 3) and females (test group 3 only) were regarded as related to treatment and adverse. Under the conditions of the study the No Observed Adverse Effect Level (NOAEL) for maternal toxicity was determined to be 300 mg/kg bw/day. The NOAEL for developmental toxicity was determined to be 300 mg/kg bw/day.

Important considerations for the use of read-across for acute toxicity are: i) 4-methyl-3-decen-5-ol (the target substance) has similar physico-chemical properties to (2E)-3,7-dimethylocta-2,6 -dien-1-ol (the source substance), ii) there are structural similarities between the two substances and iii) the OECD QSAR Toolbox assigns an identical toxicity profile to both chemicals. The source substance is therefore considered suitable for classification and labelling and risk assessment purposes using a read-across approach (see 'Justification for read-across' in Section 13 for further details).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3550 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

2-TRANS-3,7-DIMETHYL-2,6-OCTADIEN-1-OL

Cas Number:

106-24-1

IUPAC Name:

2-TRANS-3,7-DIMETHYL-2,6-OCTADIEN-1-OL

Constituent 2

Reference substance name:

Geraniol

EC Number:

203-377-1

EC Name:

Geraniol

Cas Number:

106-24-1

Molecular formula:

C10H18O

IUPAC Name:

(E)-3,7-dimethylocta-2,6-dien-1-ol

Test material form:

other: liquid, not further specified

Details on test material:

- Molecular formula (if other than submission substance): C10H18O- Molecular weight (if other than submission substance): 154.25- Smiles notation (if other than submission substance): C(C)(C)=CCCC(C)=CCO- Structural formula attached as image file (if other than submission substance): see Fig. 1- Appearance: yellowish / clear liquid- Storage condition of test material: room temperature

Test animals

Species:

rat

Strain:

other: Crl:WI(Han)

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Charles River Laboratories, Research Models and Services, Germany GmbH- Age at study initiation: approximately 11 - 12 weeks (at start of administration)- Weight at study initiation: 286.8 - 322.3 g (males); 189.8 - 219.5 g (females)- Housing: Individually in Makrolon type M III cages (Becker & Co., Castrop-Rauxel, Germany) except during mating when one male and one female were housed together in Makrolon type M III cages. Pregnant females and their litters were housed together until the end of lactation. Pregnant females were provided with nesting material (cellulose wadding) towards the end of gestation. - Use of restrainers for preventing ingestion (if dermal): no- Diet:ground KLiba maintenance diet mouse/rat "GLP" meal (Provimi Kliba SA, Kaiseraugst, Switzerland) ad libitum- Water: ad libitum- Acclimation period: 7 daysENVIRONMENTAL CONDITIONS- Temperature (°C): 20 - 24 °C- Humidity (%): 30 - 70 %- Air changes (per hr): 15 air changes per hour- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light

Administration / exposure

Route of administration:

dermal

Vehicle:

other: Corn Oil

Details on exposure:

TEST SITE- Area of exposure: Dorsal- Type of wrap if used: A semiocclusive dressing (4 layers of absorbent gauze and stretch bandage)- Time intervals for shavings or clipplings: The dorsal skin was clipped at least once a week depending on hair growth.REMOVAL OF TEST SUBSTANCE- Washing: After removal of the dressing, the application area was washed with lukewarm water.- Time after start of exposure: Performed dailyTEST MATERIAL- Amount(s) applied (volume or weight with unit): 4 mL/kg. The calculation of the volume administered was generally based on the most recent individual body weights.- Constant volume or concentration used: noVEHICLE- Amount(s) applied (volume or weight with unit): 4 mL/kgUSE OF RESTRAINERS FOR PREVENTING INGESTION: no

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analytical verification of test concentrations was performed on samples of all concentrations at the start and towards the end of the adminstration period. Concentration analysis was performed by HPLC with UV detection. The HPLC conditions were as follows:- Column: Gemini C18 5 µ 110A, 150 x 3 mm- Eluent: 50 % acetonitrile + 0.5 M sulphuric acid (5 mL/L); 50 % highly deionised water + 0.5 M sulphuric acid (5 mL/L)- Flow rate: 0.6 mL/min- Injection volume: 2, 5 µL- Column temperature: ambient- Detection: 205 nm- Limit of quantification: 12.8 mg/L

Details on mating procedure:

- M/F ratio per cage: one male and one female of the same treatment group - Length of cohabitation: animals were mated overnight (from 16:00 to 07:00 - 09:00 the following morning)- Further matings after two unsuccessful attempts: yes, animals were mated for a maximum of two weeks.- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

The duration of treatment covered the premating period of 2 weeks and mating period (maximum of 2 weeks) in both sexes, approximately 1 week post-mating in males, and the entire gestation period until gestation day 19 in females (although the females were not treated during the end of gestation and during lactation).

Frequency of treatment:

Daily (6 hours exposure per day, 7 days per week)

Duration of test:

Up to gestation day 19 for parental animals (with a two week pre-mating period and a two week (maximum) mating period), and postnatal day 4 for offspring

No. of animals per sex per dose:

10 females per dose level

Control animals:

yes

Details on study design:

- Dose selection rationale: In a previous study rats received dermal applications of test material at the dose levels 0, 300, 500, 750 and 1000 mg/kg bw/day for 14 days. Corn oil served as vehicle. In this study the test material caused scales and erythema at dose levels of 500 mg/kg bw/day and above but no findings at a dose level of 300 mg/kg bw/day. Therefore 450 mg/kg bw/day was selected as the high dose and 150 and 50 mg/kg bw/day were selected as intermediate and low doses, respectively.- Rationale for animal assignment: Animals were distributed according to weight among the individual test groups, separated by sex. The weight variation of the animals used did not exceed 20 % of the mean weight of each sex.

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes - Time schedule: mortality was checked twice daily and clinical observations were performed at least once dailyBODY WEIGHT: Yes - Time schedule for examinations: Weekly with the following exceptionsFemales were weighed on the day of positive evidence of sperm (GD 0), and on GD 7, 14 and 20 during mating.Females showing no positive sign of sperm in the vaginal smear were weighed once a week during mating as were malesFemales with litter were weighed on the day of parturition (PND 0) and on PND 4.Females without litter were weighed once a weekFemaels between PND 4 and sacrifice were weighed once a week.FOOD CONSUMPTION: Yes- Time schedule for examinations: weekly (food consumption was not determined in the females without positive evidence of sperm during mating and gestation periods and in the females without litter during lactation period) Food consumption was not determined during the mating periodFood consumption of the females with evidence of sperm was determined for GD 0-7, 7-14 and 14-20.Food consumption for females which gave birth to a litter was determined for PND 1-4.WATER CONSUMPTION: NoOTHER: The parturition and lactation behaviour of the dams was generally evaluated once daily. The day of parturition was considered to be the 24 hour period from about 15:00 of one day until about 15:00 of the following day.SACRIFICE: Parental animals were sacrificed by decapitation under isoflurane anesthesia. The exanguinated animals were necropsied and assessed by gross pathology.- Sacrifice on gestation day 19Animals which died intercurrently or were sacrificed in a moribund state were necropsied as soon as possible after their death and assessed by gross pathology.ORGAN WEIGHTSThe weights of the following were determined: carcass and ovariesORGAN/TISSUE FIXATIONThe following were fixed in 4 % neutral buffered formaldehyde solution or in modified Davidson's solution: all gross lesions, adrenal glands, ovaries, pituitary gland, coagulation glands, skin (treated), skin (untreated), uterus, oviducts, vagina.The ovaries of animals that died or were sacrificed intercurrently were fixed in 4 % buffered formaldehyde solution.HISTOPATHOLOGYAfter organs were fixed, histotechnical processing and examination by light microscopy were performed on gross lesions and treated skin of animals of all test groups and on the ovaries and untreated skin in the control and high dose group animals.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: YesExaminations included:- Number of implantations: Yes

Fetal examinations:

PARAMETERS EXAMINEDThe following parameters were examined in F1 offspring: number and sex of pups, stillbirths, live births, gross-morphological changes. On post-natal day 1 live pups were weighed.On post-natal day 4, the pups were sacrificed under isoflurane anesthesia with CO2. After sacrifice, the pups were examined externally and eviscerated, and their organs were assessed macroscopically.Pups that died or were sacrificed in a moribund state were eviscerated and examined for possible defects and/or the cause of death.All pups without any notable findings were discarded after their macroscopic evaluation.

Statistics:

Dunnett’s-test (two-sided) for the hypothesis of equal means was performed on the following parameters: food consumption; body weight; body weight change; number of mating days; duration of gestation; number of pups delivered per litter; implantation sites and post implantation loss. Pair-wise comparison of each dose group with the control group was performed with Fisher’s Exact test on the following parameters: mating indices; fertility indices; gestation index; females with liveborn and stillborn pups; live birth index, stilibrin, dead, cannibalised, sacrificed and moribund pups; viability index and number of litters with affected pups at necropsy. Pairwise comparison of each dose group was analysed with the Wilcoxon-test on the parameters: portions of affected pups per litter with necropsy observations.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

See "Details on results (parental animals)" for information

Dermal irritation (if dermal study):

effects observed, treatment-related

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Other effects:

not examined

Maternal developmental toxicity

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

not specified

Dead fetuses:

no effects observed

Changes in pregnancy duration:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed

Changes in number of pregnant:

no effects observed

Other effects:

not specified

Details on maternal toxic effects:

Maternal toxic effects:no effects. Remark: Only local signs of irritation were reportedDetails on maternal toxic effects:CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)Female no. 127 (dosed at 150 mg/kg bw/day) was sacrificed moribund on study day 11 because of a severe thoracic injury. No test material-related mortalities occurred in any test group.For female animals, different dermal findings were seen on several days. In test group 3, 7 females showed slight and moderate erythema on several days of the study, starting on study day 3. In addition, some animals of test group 3 showed multifocal, focal and diffuse scales on treated skin starting on study day 5. Similar findings but less pronounced were observed in female animals of test group 2, i.e. focal red spots, slight erythema and focal scales on tre6aed skin on several days of the study. In test group 1, slight erythema as well as focal and diffuse scales on treated skin were observed in individual animals at different time points.Female no. 137 of test group 3, which did not deliver pups, showed a vaginal haemorrhage 27 days after mating.In female animal 127 of test group 2, a severe thoracal injury was observed during pre-mating. It was a self inflicted injury which was caused by the animal's attempt to get rid of the gauze. The injury became more severe by time and the animal had to be sacrificed in a moribund state on study week 1.BODY WEIGHTThe mean body weight and body weight change of animals showed no significant deviations to the control group in all test groups.FOOD CONSUMPTION No changes in food consumption of toxicological significance were seen for male and female animals of all test groups.ORGAN WEIGHTSNone of the absolute of relative weight parameters in test groups 1 - 3 showed relevant differences when compared to the control group and were considered to be within the normal range.GROSS PATHOLOGYA red-brown lesion was noted on the thorax of female no. 127 of test group 2 which was sacrificed moribund. All gross lesions observed in test animals occurred singularly. They were considered to be spontaneous lesions in origin and not related to treatment.HISTOPATHOLOGY The lesion noted on the thorax of female no. 127 correlated to an erosion on the skin. Lymphocytic infiltrates were observed in treated skin sections which were distributed band-like between epidermis and dermis. All other findings noted were single observations which were considered to be incidental and spontaneous in origin and without any relation to treatment.

Effect levels (maternal animals)

1

Key result

Dose descriptor:

NOAEL

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

2

Key result

Dose descriptor:

NOAEL

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Results (fetuses)

Fetal body weight changes:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Visceral malformations:

no effects observed

Other effects:

not specified

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effectsDetails on embryotoxic / teratogenic effects:VIABILITY (OFFSPRING)No significant findings for Pups that died during lactation were observed. The viability index varied between 99 % for test groups 1 and 3 and 100 % for test groups 0 and 2.No test material-related changes were obtained.Male pup no. 8, delivered by dam no 118 of test group 1, was sacrificed moribund on post natal day 0 as it showed malformation of the skull, anophthalmia and cleft lip.The sex distribution and sex ratios of live F1 pups on the day of birth and post natal day 4 did not show biologically relevant differences between the control and test groups 1 to 3.CLINICAL SIGNS (OFFSPRING)Dam no 118 (test group 1) gave birth to male pup no 8 with a deformation of its snout. On first sight a cleft lip and anophthalmia were seen. The staining of the pups skull showed that several bones (basisphenoid, palatine, incisive, nasal - including cartilage and maxilla) were deformed and/or displaced. therefore, F1 pups did not show adverse clinical signs up to schedules sacrifice on post natal day 4.BODY WEIGHT (OFFSPRING)The mean body weight of female pups was increased by 10 % on post natal day 4 in test group 3. Pup body weight change in groups 1 to 3 were comparable to the concurrent control values.

Effect levels (fetuses)

Key result

Dose descriptor:

NOAEL

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no effects observed

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Any other information on results incl. tables

Homogeneity and concentration control analyses

The test material was completely miscible with Corn Oil Ph. Eur, and thus, a solution. Therefore, the test material preparation was considered to be homogenous and no further homogeneity analysis was performed.

The concentration control analyses at the beginning and towards the end of the application period revealed values in the expected range of the target concentrations. the means of the nominal concentrations were in a range of 96.5 - 104.3 % of the nominal concentrations.

ANALOGUE APPROACH JUSTIFICATION:

- See attached “Justification for read-across” document for full details.

- In summary, important considerations for the use of read-across for reproductive/developmental toxicity screening are: i) 4-methyldec-3-en-5-ol (the target chemical) has similar physico-chemical properties as 4(2E)−3,7−dimethylocta−2,6−dien−1−ol (the source substance), ii) there are structural similarities between the two chemicals and iii) the OECD QSAR Toolbox assigns an identical toxicity profiles to both chemicals.

The information reported in this summary is included to demonstrate comparability between the source (4(2E)−3,7−dimethylocta−2,6−dien−1−ol) and target (4-methyldec-3-en-5-ol) substance.

Applicant's summary and conclusion

Conclusions:

Under the conditions of the study the No Observed Adverse Effect Level (NOAEL) for maternal toxicity was determined to be 300 mg/kg bw/day. The NOAEL for developmental toxicity was determined to be 300 mg/kg bw/day.

Executive summary:

The reproductive and developmental toxicity of the test material was investigated in a GLP study which was conducted in accordance with standardised guidelines OECD 421 and EPA OPPTS 870.3550. During the study test material was administered via dermal administration to groups of 10 male and 10 female Wistar rats at dose levels of 0 (vehicle control; test group 0), 50 (test group 1), 150 (test group 2) and 450 mg/kg (test group 3) in order to observe the possible effects of the test material on the integrity and performance of the reproductive system in both sexes. Due to severe dermal findings, the dose level for test group 3 was decreased to 300 mg/kg bw/day from study day 10 onwards. Only signs of local dermal toxicity were observed for males and females at all dose levels. No changes in food consumption or body weight were seen at any dose level. fertility indices for male and female animals were not impaired by test material administration. Furthermore, there were no treatment-related or histological findings in ovaries, testes or epipdiymides associated with dermal administration of the test material. The local minimal inflammatory reactions in the skin of treated males (test groups 1 to 3) and females (test group 3 only) were regarded as related to treatment and adverse. Under the conditions of the study the No Observed Adverse Effect Level (NOAEL) for maternal toxicity was determined to be 300 mg/kg bw/day. The NOAEL for developmental toxicity was determined to be 300 mg/kg bw/day.

Important considerations for the use of read-across for acute toxicity are: i) 4-methyl-3-decen-5-ol (the target substance) has similar physico-chemical properties to (2E)-3,7-dimethylocta-2,6 -dien-1-ol (the source substance), ii) there are structural similarities between the two substances and iii) the OECD QSAR Toolbox assigns an identical toxicity profile to both chemicals. The source substance is therefore considered suitable for classification and labelling and risk assessment purposes using a read-across approach (see 'Justification for read-across' in Section 13 for further details).