(1R,5R)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Screening for reproductive/developmental toxicity: Data waiving (study scientifically not necessary): According to REACH Annex VIII, column 2, the screening study does not need to be conducted because a pre-natal developmental toxicity study is available.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because a pre-natal developmental toxicity study is available

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
According to the column 2 of REACH Annex VIII, the study does not need to be conducted because a pre-natal developmental toxicity study is available.

Reason / purpose for cross-reference:

data waiving: supporting information

Effect on fertility: via oral route

Endpoint conclusion:

no study available

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Screening for reproductive/developmental toxicity: Data waiving (study scientifically not necessary): According to REACH Annex VIII, column 2, the screening study does not need to be conducted because a pre-natal developmental toxicity study is available.

Effects on developmental toxicity

Description of key information

Developmental toxicity/Teratogenicity: Key study. Based on the experimental results obtained with the analogue substance camphene, d-alpha pinene was determined to not have teratogenic properties. The NOEL for dams and fetuses was calculated to be 250 mg/kg bw/day. Also, as no teratogenic effects were observed at highest dose tested, a NOAEL >1000 mg//kg bw/day for developmental toxicity was established.

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 April 1992 – 05 August 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Test method according to OECD guideline 414 and GLP, but only two dose levels were tested.

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

(Only two dose levels were tested)

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Lippische Versuchstierzucht, HAGEMANN GmbH, D-4923 Extertal 1, Stock: Tif: RAI f (SPF)
- Age at study initiation: ca. 54 days old
- Weight at study initiation: 180 - 190 g
- Housing: The dams were kept singly in MAKROLON cages type III.
- Diet (e.g. ad libitum): ALTROMIN 1314 (supplied by: ALTROMIN GmbH, P.O.Box 285, D-4937 Lage/Lippe) served as food, ad libitum.
- Water (e.g. ad libitum): Tap water, ad libitum.
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 ºC
- Humidity (%): 50 ± 15 %
- Photoperiod: 12 hrs dark / 12 hrs light (150 lux at 1.50 m room height)

IN-LIFE DATES: 28 April 1992 – 05 August 1992

Route of administration:

oral: gavage

Vehicle:

other: sesame oil, DAB 10

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test substance-vehicle mixtures were freshly prepared each day immediately before dosing.
The stability and homogeneity of the solution was ensured for a period of 4 hours

VEHICLE
- Concentration in vehicle: 50 mg/ml and 200 mg/ml
- Amount of vehicle (if gavage): 5 mL/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

For the determination of the concentration in the substance-vehicle preparations, 2 samples of 10 mL (each one for the concentrations 50.0 mg/Camphene/mL suspension and 200.0 mg Camphene/mL suspension) were analyzed at the start and at the end of treatment.
The determination of the concentrations was performed by GC using a FID detector.

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused: Females randomly chosen were mated overnight with fertile ('proved') 4 - 12 months old male rats of the same breed. They were repeatedly employed, at the earliest three days after successful copulation.
- M/F ratio per cage: 1 male/1 female
- Proof of pregnancy: Sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

From the 6th to 15th day of pregnancy

Frequency of treatment:

Daily

Duration of test:

20 days (from day 0 to day 20 of pregnancy)

Dose / conc.:

250 mg/kg bw/day (actual dose received)

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

20 pregnant female rats per group (plus 15 reserve animals)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
During a range-finding study using 3 pregnant rats from the same strain as selected for the present study, a camphene dose-Ievel of 1000 mg/kg b.w., by gavage, was administered from day 6 to 15 of pregnancy. 1000 mg/kg b.w. is the highest requested dose according to the OECD method 414 (limit test for embryotoxicity).
Except for a slight transient reaction after the first dosing, 1000 mg/kg b.w. was well-tolerated by the dams. 1000 mg/kg b.w. did not influence the prenatal development. Based on these results, 250 mg and 1000 mg camphene/kg b.w./day, by gavage, treatment from the 6th to 15th day of prenancy were selected by the sponsor as doses for the main experiment.

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked in table were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily checks (once in the morning and once as late on the day as practicable) were performed on behaviour, external appearance, mortality and faeces.

BODY WEIGHT: Yes
- Time schedule for examinations: daily (always at the same time in the morning)

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption was determined daily by weighing the residue.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Intake of drinking-water was observed daily

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: On the 20th day of gestation the surviving rats were laparotomised under ether narcosis. The ovaries and uteri were removed and examined. To check for possible drug effects a dissection was carried out which included macroscopic examination of internal organs. A full macroscopic inspection was carried out in the prematurely deceased dams as soon as possible after their exitus.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes. The uterus weight was determined (with and without fetuses), weight of the ovaries was determined.
- Number of corpora lutea: Yes.
- Number of implantations: Yes. Implantations and location of fetuses in the uterus were determined.
- Number of early resorptions: Yes.
- Number of late resorptions: Yes.
- Other:
Sex and viability of fetuses were determined. Animals are said to be viable when they are found alive (spontaneous breathing, spontaneous movement).
Weights and length of fetuses and weight of the placentae were determined (fetuses were considered as runts if their weight was less than 70% of the mean litter weight).

Fetal examinations:

- External examinations: Yes: [all per litter]
- Soft tissue examinations: Yes: [half per litter]. According to the method of Wilson.
- Skeletal examinations: Yes: [half per litter]. According to the method of Dawson.

Statistics:

The comparison of malformation and variation rates was carried out using R. A. FISHER's exact test (p ≤ 0.05). All other data were evaluated in the following way:
Homogeneity of variances was tested by the Bartlett chi-square test, and, if the variances were homogeneous, a one-way analysis of variance was applied. When the results indicated a significant difference among groups the DUNNETT test (p ≤ 0.01) was used to compare the experimental groups with the control group.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Six of the 20 dams treated with 1000 mg/kg bw/day showed reduced motor activity and salivation after first dosing, one of them salivation after second dosing. One more dam showed salivation on the second dosing. The reactions occurred within 5 - 20 min after administration and lasted for 20 - 60 min, 1-2 hrs or 2-6 hrs.
No clinical signs were observed in the remaining high-dosed and the low-dosed dams.

Dermal irritation (if dermal study):

not examined

Mortality:

mortality observed, non-treatment-related

Description (incidence):

Two dams treated with 1000 mg/kg bw/day died prematurely probably due to regurgitation and aspiration of part of the administered test compound. Because deaths were no substance-related these dams were replaced to have 20 pregnant females in this dose group for study examinations.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Body weight and body weight gain remained within the normal range of the controls.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Transient impairment of the food consumption by the highest tested dose (1000 mg/kg bw/day) was observed on the 7th, 8th and 9th gestation day by 6%, 22% and 10%, respectively.

Food efficiency:

not examined

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

no effects observed

Description (incidence and severity):

Treatment did not influence drinking-water consumption.

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

Six of the 20 dams treated with 1000 mg/kg bw/day showed reduced motor activity and salivation after first dosing, one of them salivation after second dosing. One more dam showed salivation on the second dosing. The reactions occurred within 5 - 20 min after administration and lasted for 20 - 60 min, 1-2 hrs or 2-6 hrs.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

The uterus weights of the treated groups were not influenced by the exposure to the test compound.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No substance-related pathological changes were detected at autopsy.
Macroscopic inspection during dissection of the two deceased rats revealed emphysematous and spongy lungs, these findings indicating regurgitation and aspiration of part of the administered test compound.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Number of abortions:

no effects observed

Description (incidence and severity):

No abortions were registered either in treated groups or in the control group.

Pre- and post-implantation loss:

effects observed, non-treatment-related

Description (incidence and severity):

There were no substance-related and/or statistically significant differences between the treated groups and the control group for the pre-implantation loss.
However, the high dose caused a slight but not significant (at p < = 0.01) increase of the post-implantation loss (substance-treated group: 11.5%, control group: 5.2%).

Total litter losses by resorption:

effects observed, non-treatment-related

Description (incidence and severity):

The mean value of total losses by resorptions per litter were calculated to be 0.8 in the control group, 1.0 in the low dose group and 1.3 in the high dose group (statistically not significant at p < = 0.01).

Early or late resorptions:

effects observed, non-treatment-related

Description (incidence and severity):

1000 mg camphene/kg b.w./day caused a slight but not significant (at p < = 0.01) increase of the resorption rate (substance-treated group: 11.5%, control group: 5.2%).

Dead fetuses:

no effects observed

Description (incidence and severity):

No dead fetuses occurred in the substance treated and control dams.
The treated groups had a number of viable fetuses per pregnant rat similar to the control group.

Changes in pregnancy duration:

not specified

Changes in number of pregnant:

no effects observed

Description (incidence and severity):

There were no substance-related and/or statistically significant differences between the treated groups and the control group in conception rate, in the mean number of corpora lutea and implantation site.
The conception rate varied between 84% and 96%.

Other effects:

no effects observed

Key result

Dose descriptor:

NOEL

Effect level:

250 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

clinical signs

Key result

Abnormalities:

no effects observed

Fetal body weight changes:

no effects observed

Description (incidence and severity):

The mean fetal weights and mean placental weights were not influenced by the test substance exposure. All values were within the range of biological variation.

Reduction in number of live offspring:

no effects observed

Description (incidence and severity):

There were no substance-related and/or statistically significant differences between the treated groups and the control group in the number of viable fetuses.

Changes in sex ratio:

no effects observed

Description (incidence and severity):

The sex distribution of the fetuses in the treated groups was comparable with the control fetuses. The differences observed in comparison to the control are without any biological relevance.

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

not examined

External malformations:

effects observed, non-treatment-related

Description (incidence and severity):

External examination of the fetuses revealed malformations (shifted and fused dorsal, lumbar and coccygeal vertebrae, bilateral crossed legs, stump tail, omphalocele) in one fetus out of 243 fetuses in the high dose group and only one variation (haemorrhage in the left anterior chamber of the eye) in one fetus out of 248 fetuses in the low dose group. One out of 258 control foetuses showed one malformation (stump tail). These changes belong to the spontaneous range as to their type and the number of affected foetuses.

Skeletal malformations:

effects observed, non-treatment-related

Description (incidence and severity):

No malformations were observed during skeletal examinations.
The variations elicited were related to the ribs (accessory 14th ribs, wavy ribs) and the sternum (sternebra bipartite or misaligned) and were found in all groups, including the control, to about the same extent.
In all groups signs of retardations (incomplete or missing ossification of hyoid, skull, vertebral bodies and/or sternebra were found without any clear differences of biological relevance between the groups, included the control).

Visceral malformations:

effects observed, non-treatment-related

Description (incidence and severity):

The examination of the organ of the foetuses revealed no malformations in the treated groups.
Variations were detected in all groups. The very common findings (uni- or bilateral dilated renal pelvis, haemorrhages of the liver, 4th cerebral ventricle enlarged) in the rat strain used in the study occurred without any dose-response relationship.
The type and number of variations found in the treated groups were similar to those found in the control group.

Other effects:

no effects observed

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Fetuses toxicity

Remarks on result:

other: No adverse effect observed at the highest dose tested

Key result

Abnormalities:

effects observed, non-treatment-related

Localisation:

other: (shifted and fused dorsal, lumbar and coccygeal vertebrae, bilateral crossed legs, stump tail, omphalocele)

Description (incidence and severity):

One malformed fetus at 1000 mg/kg bw/day belongs to the spontaneous range as to type and number of affected fetuses (control: one fetus with stump tail).

Key result

Developmental effects observed:

no

Conclusions:

Under the test conditions, the NOEL for the dams and for the fetal organism was 250 mg camphene/kg bw/day. Camphene did not possess teratogenic properties.

Executive summary:

A pre-natal developmental toxicity test was performed with camphene according to OECD Guideline 414. 20 pregnant Sprague-Dawley rats were orally exposed by gavage during the 'critical' phase of organogenesis (daily from the 6th to 15th day of pregnancy). Tested concentrations were 0, 250 and 1000 mg/kg bw/day, using sesame oil as vehicle. Their development during the gestation period was observed. On day 20 of pregnancy the animals were laparotomised and examined macroscopically for implantation sites, resorptions in the uterus and for condition of the fetuses.

 No substance-related mortality was observed in the dams. Six of the 20 dams treated with 1000 mg/kg bw/day showed reduced motor activity and salivation. No other clinical signs were observed in the remaining high-dosed and the low-dosed dams. Body weights remained within the normal range and body weight gain showed no influence of the test compound. Transient impairment of the food consumption by the highest tested dose was observed. Treatment did not influence drinking-water consumption. No substance-related pathological changes were detected at autopsy.

The highest tested dose caused a slight but not significant (at p<= 0.01) increase of the resorption rate and, consequently, of the post-implantation loss. No further influence on the prenatal development was detected. All other fetal parameters were within the normal range of the control group. External macroscopic inspection, examination of soft tissue (WILSON's method) and skeletal examination (staining of the skeleton according to DAWSON's method) revealed no substance-related variations and/or retardations. One malformed fetus at 1000 mg/kg bw/day (shifted and fused dorsal, lumbar and coccygeal vertebrae, bilateral crossed legs, stump tail, omphalocele) belongs to the spontaneous range as to type and number of affected fetuses (control: one fetus with stump tail). No dead fetuses occurred in the substance treated and control dams.

Under these test conditions, the NOEL for the dams and for the fetal organism was 250 mg camphene/kg bw/day. Camphene did not possess teratogenic properties.

Reference 2

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

03 January 1992 – 02 April 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Dose-Range-Finding study. Test method according to OECD guideline 414 and GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

(Only three animals were used per group)

GLP compliance:

yes (incl. QA statement)

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Lippische Versuchstierzucht, HAGEMANN GmbH, D-4923 Extertal 1
- Age at study initiation: ca. 71 days old
- Weight at study initiation: 211 - 245 g
- Housing: The dams were kept singly in MAKROLON cages type III.
- Diet (e.g. ad libitum): ALTROMIN 1314 (supplied by: ALTROMIN GmbH, P.O.Box 285, D-4937 Lage/Lippe) served as food, ad libitum.
- Water (e.g. ad libitum): Tap water, ad libitum.
- Acclimation period: Five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 ºC
- Humidity (%): 50 ± 15 %
- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: 03.01.1992 To: 23.01.1992

Route of administration:

oral: gavage

Vehicle:

other: sesame oil, DAB 9

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test substance-vehicle mixtures were freshly prepared each day immediately before dosing.
Volume of administration: 5 mL/kg bw

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

Fertile ('proved') 4 - 12 months old male rats of the same breed served as partners. They were repeatedly employed, at the earliest three days after successful copulation. The female breeding partners were randomly chosen.
Matings were monogamous.
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

From the 6th to 15th day of pregnancy

Frequency of treatment:

Daily

Duration of test:

20 days (from day 0 to day 20 of pregnancy)

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

3 pregnant female rats per group (plus 2 reserve animals)

Control animals:

yes, concurrent vehicle

Maternal examinations:

Daily checks (once in the morning and once as late on the day as practicable) were performed on behaviour, external appearance, mortality and faeces.
Body weight was ascertained daily - always at the same time in the morning - and these measurements were also used for calculating the daily amount of test compound to be administered.
Food consumption was determined daily by weighing the residue. Intake of drinking-water was observed daily.

Ovaries and uterine content:

On the 20th day of gestation the surviving rats were laparotomised under ether narcosis. The ovaries and uteri were removed and examined. To check for possible drug effects a dissection was carried out which included macroscopic examination of internal organs.
Fetuses were removed and the following examinations performed:
A count was taken of fetuses and placentae.
Sex and viability of fetuses were determined. Animals are said to be viable when they are found alive (spontaneous breathing, spontaneous movement).
Number and size of resorptions were determined.
Corpora lutea in the ovaries, implantations and location of fetuses in the uterus were determined.
The uterus weight was determined (with and without fetuses), weight of the ovaries was determined.
Weights and length of fetuses and weight of the placentae were determined (fetuses were considered as runts if their weight was less than 70% of the
mean litter weight).

Fetal examinations:

Fetuses were inspected externally for damages, especially for malformations.
Fetuses were dissected and the number and type of possible variations (incl. retardations) or malformations was determined macroscopically.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Two of the three dams treated with 1000 mg Camphene/kg bw showed reduced motor activity and salivation after the first dosing. These reactions occurred within 20 minutes after gavage treatment and lasted for 1-2 hours in dam nº. 5 and for 2-6 hours in dam nº. 6. No corresponding clinical signs were seen at the further observations and examinations, the third dam used remained inconspicuous. Consistency of faeces did not differ between substance-treated and control rats.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

None of the dams died prematurely.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

1000 mg Camphene/kg bw/day had no influence on body weight gain of the dams. All values remained within the range of the controls.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

1000 mg Camphene/kg bw/day had no influence on food consumption of the dams. All values remained within the range of the controls.

Food efficiency:

not examined

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

no effects observed

Description (incidence and severity):

1000 mg Camphene/kg bw/day had no influence on drinking-water consumption of the dams. All values remained within the range of the controls.

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

Two of the three dams treated with 1000 mg Camphene/kg bw showed reduced motor activity and salivation after the first dosing. These reactions occurred within 20 minutes after gavage treatment and lasted for 1-2 hours in dam nº. 5 and for 2-6 hours in dam nº. 6.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

The uterus and ovaries weights of the treated groups were not influenced by the exposure to the test compound.

Gross pathological findings:

no effects observed

Description (incidence and severity):

None of the dams showed macroscopically visible organic changes at necropsy.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Description (incidence and severity):

Number of implantations showed no abnormalities.

Total litter losses by resorption:

no effects observed

Description (incidence and severity):

Resorptions were within the normal range.

Early or late resorptions:

no effects observed

Description (incidence and severity):

Resorptions were within the normal range.

Dead fetuses:

no effects observed

Description (incidence and severity):

Macroscopic inspection during laparotomy revealed that dead foetuses did not occur.

Changes in pregnancy duration:

not specified

Changes in number of pregnant:

no effects observed

Description (incidence and severity):

Number of corpora lutea, implantations and foetuses showed no abnormalities.

Other effects:

no effects observed

Dose descriptor:

NOAEL

Effect level:

< 1 000 mg/kg bw/day

Basis for effect level:

other: maternal toxicity

Abnormalities:

no effects observed

Fetal body weight changes:

no effects observed

Description (incidence and severity):

Foetal weights and length as well as weight of the placentae did not differ substance-relatedly from the controls.

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

not specified

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

not specified

External malformations:

no effects observed

Description (incidence and severity):

Macroscopic inspection during laparotomy revealed no variations/retardations or malformed foetuses. None of the foetuses was a runt.

Skeletal malformations:

no effects observed

Description (incidence and severity):

Macroscopic inspection during laparotomy revealed no variations/retardations or malformed foetuses. None of the foetuses was a runt.

Visceral malformations:

no effects observed

Description (incidence and severity):

Macroscopic inspection during laparotomy revealed no variations/retardations or malformed foetuses. None of the foetuses was a runt.

Other effects:

no effects observed

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Basis for effect level:

other: teratogenicity

Abnormalities:

no effects observed

Developmental effects observed:

no

Conclusions:

Under the present test conditions, Camphene did not influence the prenatal development. Except for a slight transient reaction after the first dosing (reduced motor activity, salivation) the dose-level of 1000 mg/kg bw/day was well-tolerated by the dams.

Executive summary:

Pregnant Sprague-Dawley rats were orally treated with the test substance by gavage during the 'critical' phase of organogenesis (daily from the 6th to 15th day of pregnancy). Tested concentrations were 0 and 1000 mg/kg bw/day, using sesame oil as vehicle. 3 pregnant rats were used in each group. Their development during the gestation period is observed. On day 20 of pregnancy the animals were laparotomised and examined macroscopically for implantation sites, resorptions in the uterus and for condition of the fetuses.

Influence on the dam:

None of the dams died prematurely. Two of the three treated dams showed reduced motor activity and salivation after the first dosing. These reactions occurred within 20 minutes after gavage treatment and lasted for 1-2 hours in dam nº. 5 and for 2-6 hours in dam nº. 6. No corresponding clinical signs were seen at the further observations and examinations, the third dam used remained inconspicuous. Consistency of faeces did not differ between substance-treated and control rats. Test substance had no influence on body weight gain, food and drinking-water consumption of the dams. All values remained within the range of the controls. None of the dams showed macroscopically visible organic changes at necropsy.

Influence on the fetus:

Camphene did not influence the prenatal development of rats. Number of corpora lutea, implantations and foetuses showed no abnormalities, the resorptions were within the normal range. Foetal weights and length as well as weight of the placentae did not differ substance-relatedly from the controls. Macroscopic inspection during laparotomy revealed no variations/retardations, malformed or dead foetuses did not occur. None of the foetuses was a runt.

Under the present test conditions, Camphene did not influence the prenatal development at 1000 mg/kg bw/day. Except for a slight transient reaction after the first dosing (reduced motor activity, salivation) this dose-level was well-tolerated by the dams.

Reference 3

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
The analogue substance camphene which shares the same functional groups with the target substance d-alpha pinene also has comparable values for the relevant molecular properties.
See attached the reporting format.

Reason / purpose for cross-reference:

read-across source

Species:

rat

Description (incidence):

None of the dams died prematurely.

Details on maternal toxic effects:

Based on the experimental results obtained with camphene in rat (NOAEL <1000 mg/kg bw/day for maternal toxicity), the read-across approach is applied and the NOAEL for maternal toxicity of d-alpha pinene is calculated to be <1000 mg/kg bw/day.

Dose descriptor:

NOAEL

Effect level:

< 1 000 mg/kg bw/day

Based on:

other: Read-across from an analogue

Basis for effect level:

other: maternal toxicity

Remarks on result:

other: read-across from an analogue for which NOAEL <1000 mg/kg bw

Abnormalities:

no effects observed

Details on embryotoxic / teratogenic effects:

Based on the experimental results obtained with camphene in rat (NOAEL >= 1000 mg/kg bw/day for developmental toxicity), the read-across approach is applied and the NOAEL for developmental toxicity of d-alpha pinene is calculated to be equal or greater than 1000 mg/kg bw/day.

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

other: Read-across from an analogue

Basis for effect level:

other: teratogenicity

Remarks on result:

other: Read-across from an analogue for which no adverse effect was observed at the highest dose tested

Abnormalities:

no effects observed

Developmental effects observed:

no

Conclusions:

Based on read-across approach from the analogue camphene, d-alpha pinene was determined to not influence the prenatal development at 1000 mg/kg bw/day. This dose level is expected to be well-tolerated for the dams.

Executive summary:

Pregnant Sprague-Dawley rats were orally treated with the test substance by gavage during the 'critical' phase of organogenesis (daily from the 6th to 15th day of pregnancy). Tested concentrations were 0 and 1000 mg/kg bw/day, using sesame oil as vehicle. 3 pregnant rats were used in each group. Their development during the gestation period is observed. On day 20 of pregnancy the animals were laparotomised and examined macroscopically for implantation sites, resorptions in the uterus and for condition of the fetuses.

Influence on the dam:

None of the dams died prematurely. Two of the three treated dams showed reduced motor activity and salivation after the first dosing. These reactions occurred within 20 minutes after gavage treatment and lasted for 1-2 hours in dam nº. 5 and for 2-6 hours in dam nº. 6. No corresponding clinical signs were seen at the further observations and examinations, the third dam used remained inconspicuous. Consistency of faeces did not differ between substance-treated and control rats. Test substance had no influence on body weight gain, food and drinking-water consumption of the dams. All values remained within the range of the controls. None of the dams showed macroscopically visible organic changes at necropsy.

Influence on the fetus:

Camphene did not influence the prenatal development of rats. Number of corpora lutea, implantations and foetuses showed no abnormalities, the resorptions were within the normal range. Foetal weights and length as well as weight of the placentae did not differ substance-relatedly from the controls. Macroscopic inspection during laparotomy revealed no variations/retardations, malformed or dead foetuses did not occur. None of the foetuses was a runt.

Under the present test conditions, Camphene did not influence the prenatal development at 1000 mg/kg bw/day. Except for a slight transient reaction after the first dosing (reduced motor activity, salivation) this dose-level was well-tolerated by the dams.

Based on these results, the read-across approach was applied and d-alpha pinene was determined to not influence the prenatal development at 1000 mg/kg bw/day. Except for a slight transient reaction after the first dosing (reduced motor activity, salivation) this dose-level is expected to be well-tolerated by the dams.

Reference 4

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
The analogue substance camphene which shares the same functional groups with the target substance d-alpha pinene also has comparable values for the relevant molecular properties.
See attached the reporting format.

Reason / purpose for cross-reference:

read-across source

Details on maternal toxic effects:

Based on the experimental results obtained with camphene in rat (NOEL = 250 mg/kg bw/day for maternal toxicity), the read-across approach is applied and the NOEL for maternal toxicity of d-alpha pinene is calculated to be 250 mg/kg bw/day.

Key result

Dose descriptor:

NOEL

Effect level:

250 mg/kg bw/day

Based on:

other: Read-across from an analogue

Basis for effect level:

clinical signs

Remarks on result:

other: read-across from an analogue for which NOEL = 250 mg/kg bw

Key result

Abnormalities:

no effects observed

Details on embryotoxic / teratogenic effects:

Based on the experimental results obtained with camphene in rat (NOAEL >= 1000 mg/kg bw/day for developmental toxicity), the read-across approach is applied and the NOAEL for developmental toxicity of d-alpha pinene is calculated to be equal or greater than 1000 mg/kg bw/day.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

other: Read-across from an analogue

Sex:

male/female

Basis for effect level:

other: Fetuses toxicity

Remarks on result:

other: Read-across from an analogue for which no adverse effect was observed at the highest dose tested

Key result

Abnormalities:

effects observed, non-treatment-related

Localisation:

other: (shifted and fused dorsal, lumbar and coccygeal vertebrae, bilateral crossed legs, stump tail, omphalocele)

Description (incidence and severity):

Read across from an analogue for which one malformed fetus at 1000 mg/kg bw/day was found which was considered to belong to the spontaneous range as to type and number of affected fetuses (control: one fetus with stump tail).

Key result

Developmental effects observed:

no

Conclusions:

Based on read-across approach from the analogue camphene, d-alpha pinene was determined to not have teratogenic properties. The NOEL for dams and fetuses was calculated to be 250 mg/kg bw/day.

Executive summary:

A pre-natal developmental toxicity test was performed with camphene according to OECD Guideline 414. 20 pregnant Sprague-Dawley rats were orally exposed by gavage during the 'critical' phase of organogenesis (daily from the 6th to 15th day of pregnancy). Tested concentrations were 0, 250 and 1000 mg/kg bw/day, using sesame oil as vehicle. Their development during the gestation period was observed. On day 20 of pregnancy the animals were laparotomised and examined macroscopically for implantation sites, resorptions in the uterus and for condition of the fetuses.

 No substance-related mortality was observed in the dams. Six of the 20 dams treated with 1000 mg/kg bw/day showed reduced motor activity and salivation. No other clinical signs were observed in the remaining high-dosed and the low-dosed dams. Body weights remained within the normal range and body weight gain showed no influence of the test compound. Transient impairment of the food consumption by the highest tested dose was observed. Treatment did not influence drinking-water consumption. No substance-related pathological changes were detected at autopsy.

The highest tested dose caused a slight but not significant (at p<= 0.01) increase of the resorption rate and, consequently, of the post-implantation loss. No further influence on the prenatal development was detected. All other fetal parameters were within the normal range of the control group. External macroscopic inspection, examination of soft tissue (WILSON's method) and skeletal examination (staining of the skeleton according to DAWSON's method) revealed no substance-related variations and/or retardations. One malformed fetus at 1000 mg/kg bw/day (shifted and fused dorsal, lumbar and coccygeal vertebrae, bilateral crossed legs, stump tail, omphalocele) belongs to the spontaneous range as to type and number of affected fetuses (control: one fetus with stump tail). No dead fetuses occurred in the substance treated and control dams. Under these test conditions, the NOEL for the dams and for the fetal organism was 250 mg camphene/kg bw/day. Camphene did not possess teratogenic properties.

Based on these results, the read-across approach was applied and d-alpha pinene was determined to not have teratogenic properties and the NOEL for dams and fetuses was calculated to be 250 mg/kg bw/day.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The key study has a Klimisch score = 2

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Developmental toxicity/Teratogenicity: Key study: A pre-natal developmental toxicity test was performed with camphene according to OECD Guideline 414. 20 pregnant Sprague-Dawley rats were orally exposed by gavage during the 'critical' phase of organogenesis (daily from the 6th to 15th day of pregnancy). Tested concentrations were 0, 250 and 1000 mg/kg bw/day, using sesame oil as vehicle. Their development during the gestation period was observed. On day 20 of pregnancy the animals were laparotomised and examined macroscopically for implantation sites, resorptions in the uterus and for condition of the fetuses.

No substance-related mortality was observed in the dams. Six of the 20 dams treated with 1000 mg/ kg bw/day showed reduced motor activity and salivation. No other clinical signs were observed in the remaining high-dosed and the low-dosed dams. Body weights remained within the normal range and body weight gain showed no influence of the test compound. Transient impairment of the food consumption by the highest tested dose was observed. Treatment did not influence drinking-water consumption. No substance-related pathological changes were detected at autopsy.

The highest tested dose caused a slight but not significant (at p<= 0.01) increase of the resorption rate and, consequently, of the post-implantation loss. No further influence on the prenatal development was detected. All other fetal parameters were within the normal range of the control group. External macroscopic inspection, examination of soft tissue (WILSON's method) and skeletal examination (staining of the skeleton according to DAWSON's method) revealed no substance-related variations and/or retardations. One malformed fetus at 1000 mg/kg bw/day (shifted and fused dorsal, lumbar and coccygeal vertebrae, bilateral crossed legs, stump tail, omphalocele) belongs to the spontaneous range as to type and number of affected fetuses (control: one fetus with stump tail). No dead fetuses occurred in the substance treated and control dams. Under these test conditions, the NOEL for the dams and for the fetal organism was 250 mg camphene/kg bw/day. Camphene did not possess teratogenic properties. Based on these results, the read-across approach was applied and d-alpha pinene was determined to not have teratogenic properties and the NOEL for dams and fetuses was calculated to be 250 mg/ kg bw/day.

Also, as no teratogenic effects were observed at highest dose tested, a NOAEL >1000 mg//kg bw/day for developmental toxicity can be established.

Justification for classification or non-classification

Based on the available information, the substance is not classified for toxicity to reproduction in accordance with CLP Regulation (EC) no 1272/2008.