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Effects on fertility

Link to relevant study records
Reference
Endpoint:
toxicity to reproduction
Remarks:
other: 20-weeks repeated dose toxicity study
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Reliability 2 is assigned because the information is based on a reliable subchronic repeated dose toxicity study predating OECD Guidelines and GLP
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study
Qualifier:
according to guideline
Guideline:
other:
Deviations:
yes
Remarks:
Most parameters have been evaluated according to current guidelines but not all.
Principles of method if other than guideline:
See repeated dose toxicity section
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Osborne-Mendel
Sex:
male/female
Details on test animals or test system and environmental conditions:
See repeated dose toxicity section.
Route of administration:
oral: feed
Details on exposure:
See repeated dose toxicity section.
Details on mating procedure:
Not applicable
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
See repeated dose toxicity section.
Duration of treatment / exposure:
20-weeks
Frequency of treatment:
Dietary dosing
Details on study schedule:
See repeated dose toxicity section.
Remarks:
Doses / Concentrations:

Basis:
nominal in diet
1000, 2500 and 10000 ppm
No. of animals per sex per dose:
10/sex/dose
Parental animals: Observations and examinations:
See repeated dose toxicity section.
Oestrous cyclicity (parental animals):
No information
Sperm parameters (parental animals):
No information
Litter observations:
Not applicable
Postmortem examinations (parental animals):
See repeated dose toxicity section.
Postmortem examinations (offspring):
Not applicable
Statistics:
See repeated dose toxicity section.
Reproductive indices:
Not applicable
Offspring viability indices:
Not applicable
Clinical signs:
no effects observed
Description (incidence and severity):
See repeated dose toxicity section.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
See repeated dose toxicity section.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
See repeated dose toxicity section.
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
See repeated dose toxicity section.
Other effects:
effects observed, treatment-related
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
not specified
Dose descriptor:
NOAEL
Effect level:
>= 400 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects were seen on male and female gonads in a 20-week dietary study similar to OECD TG 408, which leads to the conclusion that there are no effects on male and female fertility
Clinical signs:
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Reproductive effects observed:
not specified
Conclusions:
No effects were seen on male and female gonads after 20-weeks of administration of the test substance
Executive summary:

See repeated dose toxicity section.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Study duration:
subchronic
Species:
rat
Quality of whole database:
For alpha-Terpinyl Acetate subchronic repeated dose information is available. In addition also fertility information can be derived from its main metabolite alpha-Terpineol (being the main constituent of Terpineol multi) These are sufficiently adequate to cover the endpoint.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Alpha-Terpinyl Acetate

In the key study, alpha-Terpinyl Acetate was administered to Osborne-Mendel rats (10/dose/sex) for 20 -weeks via diet at doses of 0 (control), 10000, 2500 and 1000 ppm. For details on repeated systemic toxicity see the repeated dose section; here only information important for fertility are presented. Detailed microscopic examinations were done on the animals from the high dose group and the control group. No effect on organ weights and no macroscopic or microscopic change in the tissues in the 10000 ppm exposure group were observed including female and male gonads. These dietary doses were not converted into mg/kg bw in the publication. Therefore the ECHA guidance was used, according to R8 table R 8 -17, rats eat 40 to 50 mg/kg bw/day. 10000 ppm is equivalent to 1% in the diet. Consequently, rats exposed to 10000 ppm in the diet consumed between 400 and 500 mg alpha-Terpinyl Acetate per kg bodyweight per day. Therefore, the NOAEL for effects on gonads was calculated to be 400 mg/kg bw.

Terpineol multi

The fertility effects of the Terpineol multi being used for read across are presented as supporting. The study is presented in detail because of the effects seen in the first study but which have been disregarded based on additional testing in the second study and information on alpha-Terpinyl Acetate.

Supporting study 1A (this information is also presented in the repeated dose section). In a GLP study conducted according to OECD guideline 422, three groups, each comprising ten male and ten female rats for the Main (reproductive) phase (exception: five males at control and top dose; additional Recovery phase males were also used for pairing with Main reproductive phase females) and five female rats for the Toxicity phase received Terpineol multi at doses of 60, 250 or 750 mg/kg bw/day at a dose volume of 5 mL/kg bw/day (Thacker 2010a).

Main phase males and Toxicity phase females were dosed daily for a minimum of five consecutive weeks. An additional ten males and ten females were dosed with the vehicle or at 750 mg/kg/day for five weeks and then given two weeks of recovery before termination. Main phase females were dosed daily for two weeks before pairing, throughout mating, gestation and until Day 6 of lactation. A similarly constituted Control group received the vehicle, corn oil, at the same volume-dose. During the study, data was recorded on clinical condition, performance under detailed physical and arena examination, sensory reactivity, grip strength, motor activity, bodyweight, food consumption, water consumption (visual), haematology, blood chemistry, oestrous cycles, mating performance and fertility and gestation length. Organ weight, macroscopic and microscopic pathology investigations were undertaken in the adults. The clinical condition of offspring, litter size and survival, sex ratio and offspring bodyweight were assessed and macroscopic pathology investigations were undertaken.

Results: In the 60 mg/kg bw dose group, one male was found dead on Day 31 of study and one female was killed because of parturition difficulties. In the absence of any other death in the intermediate and high dose groups these deaths are not attributed to the test material. No significant findings were recorded for clinical signs, detailed physical examination and arena observations. Underactive behaviour and unsteady reactions, in males and females were observed briefly during Week 1 in animals receiving 750 mg/kg bw and dose‑related increases in post‑dosing salivation and chin rubbing were seen. Behavioural testing during Week 5 of dosing, including sensory reactivity findings, grip strength values and motor activity scores showed no differences considered to be associated with exposure to the test material. There were no clear effects on bodyweight in males or unmated females receiving up to 750 mg/kg bw. Males receiving 750 mg/kg bw showed lower overall weight gain (Week 0-5) compared with Control. Bodyweight during the recovery phase was similar to Controls.

Bodyweight and bodyweight gain were unaffected during gestation. During lactation females receiving 250 mg/kg bw showed lower weight gain than Controls. There were no adverse effects on food consumption in males, unmated females or females during gestation and lactation but visual assessment of water consumption indicated that males and females receiving 750 mg/kg bw were consuming more water than the Controls during the dosing period.

There were no effects of Terpineol multi on oestrous cycles, precoital interval or mating. Gestation length was within the normal range but there was a small increase in the numbers of animals at 250 mg/kg bw having longer (23 day) gestation periods. At dose levels up to and including 250 mg/kg bw there were no effects of the test material on the number of implantations, post implantation survival index, live birth index, viability index and lactation index. Male and female offspring bodyweights were not adversely affected by Terpineol multi. At 750 mg/kg bw, relative liver weights were significantly higher than Control in males and females and relative kidney weights were significantly higher than Control in males. Testis weight was markedly low in males receiving 750 mg/kg bw and there was also an indication of low epididymal weights at this dose.

Liver and kidney weights returned to normal after two weeks when the animals did not receive Terpineol multi but testis and epididymal weights showed no evidence of recovery. Adaptive centrilobular hepatocyte hypertrophy in the liver of females dosed with Terpineol multi at 750 mg/kg bw was not present after 2 weeks recovery and histopathological findings in the kidneys of males receiving 250 and 750 mg/kg bw also resolved after the end of dosing. At 750 mg/kg bw, reduced numbers or complete absence of spermatozoa, accompanied by the presence of degenerate spermatogenic cells in duct(s) were observed in the epididymides and were still present following the 2‑week recovery period. Spermatocele granuloma(ta) that were seen in two males receiving 750 mg/kg bw and one receiving 60 mg/kg/day were not seen at the end of the recovery period. The significance of this change in the single male receiving 60 mg/kg bw is uncertain as spermatocele granuloma(ta) can occur spontaneously in rats of this age and considering the absence of other degenerative changes in the testes or epididymides of this animal. Moderate to severe seminiferous tubular atrophy/degeneration was seen in the testes of all animals dosed with Terpineol multi at 750 mg/kg bw, accompanied by minimal to moderate spermatid giant cells and minimal to slight seminiferous tubular vacuolation.

Similar findings were still evident following the 2‑week recovery period but at a lower incidence and severity suggesting a degree of recovery. Based on the findings in this study, the NOAEL for males was 250 mg/kg bw. For dams the NOAEL was set at >=750 mg/kg/day.

Supporting study 1B: In the follow up study, the toxicity of Terpineol multi to the male reproductive system was examined when administered by dietary or oral gavage routes (Thacker 2010c). Three groups of Crl: CD(SD) male rats (five/dose) were dosed daily with Terpineol multi by dietary and/or oral gavage routes at the following doses: group 1: dietary 7500 ppm + supplementary gavage dose 300 mg/kg/day, group 2: dietary 10000 ppm + supplementary gavage dose 150 mg/kg/day, group 3: Terpineol multi at 750 mg/kg/day by gavage only. During the study, data was recorded on mortality, clinical condition, body weight and food consumption. Surviving animals were subjected to a detailed sperm analysis. Testes (L&R), epididymis (L&R), prostate and seminal vesicles were weighed at necropsy and tissues of right testes and epididymis were fixed for histopathological examination. During week 1 of the study, Group 1 and 2 animals consumed less diet than expected; From Day 11 Group 1 animals, receiving 7500 ppm, also received two daily doses of 150 mg/kg/b. i. d. four to five hours apart and average intake was boosted to 663 mg/kg/day whilst Group 2 animals, receiving 10000 ppm, also received a single daily dose of 150 mg/kg boosting intake to 678 mg/kg/day. Clinical signs were generally minimal. Dosing signs for Group 3 animals included salivation and chin rubbing. Food consumption for Group 1 and 2 animals receiving the test material in the diet was low throughout the study; this was attributed to the palatability of the test material. Bodyweight gains of these animals were also low. Food consumption and bodyweight changes of Group 3 animals were considered to be not adversely affected by the test material. Necropsy data indicated that decreases in reproductive organ weights and changes to macroscopic appearance were most marked in the animals receiving Terpineol multi at 750 mg/kg/day. Occasional animals (1/5 and 1/5 in each of the other groups 1 and 2 respectively) also showed changes to testicular and/or epididymal tissue appearances and/or weights. Sperm analysis showed that motile sperm with normal morphology were present in 4/5 males of Group 2 and 1/5 males of Group 1. The outliers in each group were at the extreme of achieved overall exposure for the group suggesting that absolute exposure was important, although the route of exposure and consequently potential to exceed threshold levels was of greater significance. Microscopic examination indicated there were relatively fewer changes in the testes and epididymides in the animals which were given Terpineol multi by the dietary route with oral gavage supplementation (Groups 1 and 2), whereas there were significant changes in those which received it solely by oral gavage (Group 3). The results of dietary administration suggest that exposure via the dietary route of administration reduces the testicular and sperm toxicity of the test material compared to dosing by oral gavage. The results of this study, support the hypothesis that a high peak plasma level is necessary to induce the observed toxic effects.

In conclusion, alpha-Terpinyl Acetate has no effects on fertility at the highest dose tested resulting in a NOAEL of >=400 mg/kg bw, because the fertility effects of Terpineol multi seen in the males can be disregarded because the effect is due to gavage dosing.


Short description of key information:
In the 20-weeks dietary study no effects on male and female gonads were seen at 400 mg/kg bw for alpha-Terpinyl Acetate resulting in a NOAEL for fertility for both females and males of >=400 mg/kg bw.
Information on Terpineol multi is also present on fertility (OECD TG 422 via gavage and 14-day repeated dose testing using dietary dosing). Terpineol multi can be used for read across to alpha-Terpinyl Acetate because alpha-Terpineol (the main constituent of Terpineol multi) will be the metabolites to consider during systemic exposure. Though in the OECD TG 422 testicular effects on rats were seen, these effects have been shown to be due to gavage dosing because these effects were not seen during dietary dosing. Therefore these testicular effects of Terpineol multi are not considered for the NOAEL derivation for fertility.The NOAEL for alpha-Terpinyl Acetate for males and females is therefore set to >=400 mg/kg bw.

Justification for selection of Effect on fertility via oral route:
For alpha-Terpinyl Acetate subchronic repeated dose toxicity information is used for assessing the fertility because this information has been derived from a test with a longer exposure period compared to the read across substance and because of the dietary dosing during this test, resembling the human exposure better compared to dosing via gavage.

Effects on developmental toxicity

Description of key information
The developmental toxicity of alpha-Terpinyl Acetate has been derived based on the repeated dose (oral gavage) toxicity study with reproductive/developmental toxicity screening test (OECD 422, GLP) of Terpineol multi in which no developmental toxicity effects were seen at 250 mg/kg bw, the highest dose with pregnant dams. In addition, the likelihood of other developmental toxic effects such as malformations is not expected due to e. g. the absence of structural alerts for malformation and genotoxicity.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
From April 28 to September 03, 2010
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Reliability 2 is assigned because the information is used for read across. The information as such is a GLP study conducted according to OECD guideline 422.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
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: Charles River, UK
- Age at study initiation: 9 to 10 weeks
- Weight at study initiation: 303 to 375 g for males and 198 to 253 g for females
- Housing: Up to 5 during pre-mating for all animals and after mating for males and during toxicity phase for unmatted females, individually with litter for females during gestation and lactation.
- Diet (e.g. ad libitum): Standard rodent diet (SDS VRF1 Certified) ad libitum, except overnight before routine blood sampling for Main phase males, Toxicity phase females and Recovery phase animals.
- Water (e.g. ad libitum): Potable water taken from the public supply, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25
- Humidity (%): 40 to 70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 28 April 2010 To: 29 June 2010
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Approximately 50% of the final volume of corn oil was added to the required amount of test material. The formulation was mixed using a magnetic stirrer until all of the test material had dissolved and more corn oil was added to make up the required volume. The formulation was then mixed using a magnetic stirrer until homogeneous. Initially all formulations were prepared freshly on the day of use and used within two hours of completion of preparation. However, following confirmation of the results from a homogeneity and stability, formulations were prepared weekly, subdivided into daily aliquots and used within 8 days of preparation.

VEHICLE
- Concentration in vehicle: 12 and 50 mg/mL
- Amount of vehicle (if gavage): constant dosage-volume of 5 mL/kg bw/day
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples of each formulation prepared for administration in the first and last weeks of the dosing period were analysed for achieved concentration of the test substance. Four samples were taken (nominally 1 mL accurately weighed) from all groups. Two assays from each group were analysed. The mean concentrations of CAS 8000-41-7 in test formulations analysed for the study were within applied limits, +10%/-15% of nominal concentrations, confirming accurate formulation.
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: Up to 2 weeks
- Proof of pregnancy: Vaginal plug and sperm in vaginal smear referred to as day 0 of gestation
- After successful mating each pregnant female was caged individually
- For 15 days before pairing (including the day of pairing), daily vaginal smears (dry) were taken from all Reproductive subgroup females, using cotton swabs moistened with saline. The smears were subsequently examined to establish the duration and regularity of the oestrous cycle. After pairing with the male, smearing was continued using pipette lavage, until evidence of mating was observed.
Duration of treatment / exposure:
Toxicity phase females were dosed daily for a minimum of five consecutive weeks. Main phase females were dosed daily for two weeks before pairing, throughout mating, gestation and until Day 6 of lactation. A similarly constituted Control group received the vehicle, corn oil, at the same volume-dose. Offspring were not dosed.
Frequency of treatment:
Once a day, 7 days a week
Remarks:
Doses / Concentrations:
60 and 250 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
- Reproductive subgroup (main phase): 10 females/dose
- Toxicity subgroup: 5 females/dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: In a two week preliminary study (Huntingdon Life Sciences Study No. OAD0003) which tested dose levels of 150, 600 and 1000 mg/kg bw/day. In that study animals dosed at 600 and 1000 mg/kg bw/day showed post dose observations of salivation and chin rubbing and some females at 1000 mg/kg bw/day also showed isolated incidences of reduced activity, reduced body tone and unsteady gait. An initial reduction in bodyweight was recorded in males at 600 and 1000 mg/kg bw/day. At 1000 mg/kg bw/day increased water consumption was recorded and at necropsy liver weights was increased whilst the testis and epididymal weight were reduced (67 and 76% of control, respectively).
Maternal examinations:
CAGE SIDE OBSERVATIONS:
Animals and cages were inspected visually at least twice daily for evidence of ill-health or reaction to treatment. During the acclimatisation period, observations of the animals and their cages were recorded at least once per day.

DETAILED CLINICAL OBSERVATIONS:
Detailed observations were recorded in relation to dose administration. For the Toxicity phase females dosing observations were recorded daily during the first week of treatment, twice weekly during Weeks 2 to 4 (middle and end of each week) and on one occasion during Week 5. For Main phase females these were recorded daily during the first week of dosing, twice weekly during Week 2 of dosing, on Days 0, 7, 14 and 20 after mating and on Days 1 and 7 of lactation. Observations were recorded at the following times in relation to dose administration: Pre-dose, On return of the animal to its home cage, On completion of dosing of each group, Between one and two hours after completion of dosing of all groups, As late as possible in the working day. Before treatment commenced and during each week of treatment, detailed physical examination and arena observations were performed on each animal (physical condition and behaviour during handling with particular attention to possible signs of neurotoxicity). For the Reproductive subgroup females during the post-mating period, these observations were conducted on Days 0, 7, 14 and 20 after mating and on Days 1 and 7 of lactation. A weekly physical examination including arena observations was performed during the recovery period.

BODY WEIGHT:
The weight of the Toxicity phase females was recorded on the day that dosing commenced (Week 0), weekly throughout the dosing and recovery periods and before necropsy. Main phase females were weighed on the day that dosing commenced (Week 0), weekly until mating was detected, on Days 0, 7, 14 and 20 after mating and on Days 1, 4 and 7 of lactation.

FOOD CONSUMPTION:
The weight of food supplied to each cage, that remaining and an estimate of any spilled was recorded on a weekly basis from the start of study for Toxicity phase females and Main phase females until the animals were paired for mating. Food consumption was recorded weekly (g/animal/week) during the recovery period. From these records the mean weekly consumption per animal (g/animal/week) was calculated for each cage. Food consumption was not recorded for Main phase females during pairing. For each Main phase female after mating, the weight of food supplied, that remaining and an estimate of any spilled was recorded for the periods Days 0-6, 7-13 and 14-19 after mating and Days 1-3 and 4-6 of lactation.

WATER CONSUMPTION:
Fluid intake was assessed by daily visual observation.

OTHER:
- SENSORY REACTIVITY:
Sensory reactivity and grip strength assessments were performed (before dosing) on the toxicity phase (Groups 5 and 6)/recovery phase (Control and Group 7) females during Week 5 of study. The following measurements, reflexes and responses were recorded: approach response, touch response, auditory startle reflex, tail pinch response and grip strength.

- MOTOR ACTIVITY:
During Week 5 of study (before dosing), the motor activity of the toxicity phase (Groups 5 and 6)/recovery phase (Control and Group 7) females was measured using a Rodent Activity Monitoring System (Version 2.0.3). Animals were tested individually in clear polycarbonate cages and motor activity was measured by counting infra-red beam breaks over ten 6-minute intervals (one hour total).

- HAEMATOLOGY:
During Week 5 of treatment (before dosing on each occasion) and after 2 weeks of recovery, blood samples were obtained from the toxicity phase females after overnight withdrawal of food. Animals were held under light general anaesthesia induced by isoflurane and blood samples were withdrawn from the sublingual vein. The following were measured using a Bayer Advia 120 haematology analyser: Haematocrit (Hct), Haemoglobin concentration (Hb), Erythrocyte count (RBC), Mean cell haemoglobin (MCH), Mean cell haemoglobin concentration (MCHC), Mean cell volume (MCV), Total leucocyte count (WBC), Differential leucocyte count (Neutrophils (N), Lymphocytes (L), Eosinophils (E), Basophils (B), Monocytes (M), Large unstained cells (LUC)) and Platelet count (Plt). The most common morphological changes, anisocytosis, micro/macrocytosis and hypo/hyperchromasia were recorded. Prothrombin time (PT) (using an ACL 3000 Plus analyser and IL PT-Fibrinogen reagent) and Activated partial thromboplastin time (APTT) (using an ACL 3000 Plus Analyser and IL APTT reagent) were also measured.

- BLOOD CHEMISTRY:
At the same time and using the same animals as for peripheral haematology, further blood samples (nominally 0.7 mL) were collected and the plasma was examined using a Roche P Modular Analyser for: Alkaline phosphatase (ALP), Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Gamma-glutamyl transpeptidase (gGT), Total bilirubin (Bili), Bile Acids (BIAC), Urea, Creatinine (Creat), Glucose (Gluc), Total cholesterol (Chol), Sodium (Na), Potassium (K), Chloride (Cl), Calcium (Ca), Inorganic phosphorus (Phos), Total protein (Total Prot) and Albumin (Alb) (by chemical assay).

- SACRIFICE:
Toxicity phase females were killed in Week 6 after completion of the Week 5 investigations. Main phase females (Groups 1, 5 and 6) were killed on Day 7 of lactation. Main phase females that did not litter (Group 7) were killed on Day 25 after mating. Offspring were killed on Day 7 of age.

- GROSS NECROPSY:
All animals were subject to a detailed necropsy. For Reproductive subgroup females, the number of uterine implantation sites was also recorded.

- HISTOPATHOLOGY / ORGAN WEIGHTS:
The tissues indicated in Table 1 were weighed.
The following tissues from Toxicity phase female, Main phase females that did not litter (Group 7) and those animals killed or dying prematurely were fixed for histopathology: Adrenal glands, Brain, Pituitary, Caecum, Colon, Rectum, Sciatic nerves, Duodenum, Skeletal muscle, Skin, Mammary glands (inguinal area), Heart, Spinal cord, Ileum, Spleen, Jejunum, Sternum with marrow, Kidneys, Stomach, Liver, Lungs, Thymus, Lymph nodes (mandibular and mesenteric), Thyroid with parathyroids, Trachea, Urinary bladder, Oesophagus, Uterus with cervix and oviducts, Peyer’s patch, Ovaries (L&R) and Vagina. The following tissues from each Main phase female that did litter (Groups 1, 5 and 6) were fixed for histopathology: Ovaries (L&R), Uterus with cervix and oviducts and vagina. Samples of any abnormal tissues were also retained and processed for examination.
Fetal examinations:
PARAMETERS EXAMINED:
All litters were examined at approximately 24 hours after birth and then daily thereafter for clinical signs (evidence of ill health or reaction to treatment), litter size (mortality and consequent changes in litter size from Days 1-7 of age), sex ratio of each litter (recorded on Days 1, 4 and 7 of age) and individual bodyweight (recorded on Days 1, 4 and 7 of age).

GROSS EXAMINATION OF PUPS:
All offspring killed or dying prior to scheduled termination, and of those killed at the end of the study were subjected to detailed necropsy.
Statistics:
The following sequence of statistical tests was used for grip strength, motor activity, bodyweight, food consumption, organ weight, litter size and survival indices and clinical pathology data:
1) a parametric analysis was performed if Bartlett's test for variance homogeneity (Bartlett 1937) was not significant at the 1% level. For comparisons involving two groups only t-tests were used, for all other comparisons the F1 approximate test was applied. If the F1 approximate test for monotonicity of dose-response was not significant at the 1% level, Williams' test for a monotonic trend was applied. If the F1 approximate test was significant, suggesting that the dose response was not monotone, Dunnett's test (Dunnett 1955, 1964) was performed instead.
2) a non-parametric analysis was performed if Bartlett's test was still significant at the 1% level following both logarithmic and square-root transformations. For comparisons involving two groups only, Wilcoxon’s rank sum tests (Wilcoxon 1945) were used. For all other comparisons the H1 approximate test, the non-parametric equivalent of the F1 test described above, was applied. If the H1 approximate test for monotonicity of dose-response was not significant at the 1% level, Shirley's test for a monotonic trend was applied. If the H1 approximate test was significant, suggesting that the dose-response was not monotone, Steel's test (Steel 1959) was performed instead.
For organ weight data, analysis of covariance was performed. Sex ratio were analysed by Wald chi-square test.
For gestation length estimates an exact two-tailed Linear-by-linear test (Cytel 1995), with equally spaced scores, was applied to all groups.
Indices:
REPRODUCTIVE INDICES:
Percentage mating : Number animals mating / Animals paired × 100
Conception rate (%) : Number animals achieving pregnancy / Animals mated × 100
Fertility index (%) : Number animals achieving pregnancy / Animals pairing × 100

OFFSPRING VIABILITY INDICES
Gestation index (%) : Number of live litters born / Number pregnant × 100
Post - implantation survival index (%) : Total number offspring born / Total number uterine implantation sites × 100
Live birth index (%) : Number live offspring on Day 1 after littering / Total number of offspring born × 100
Viability index (%) : Number live offspring on Day 4 after littering / Number live offspring on Day 1 after littering × 100
Lactation index (%) : Number live offspring on Day 7 after littering / Number live offspring on Day 1 after littering × 100
Percentage of males : Number of males in litter/ Total number of offspring in litter x 100
Details on maternal toxic effects:
Details on maternal toxic effects:
MORTALITY:
In the 60 mg/kg/day dose group, one female was killed because of parturition difficulties. This female had given birth to three pups and but still had 15 live pups and one early resorption in utero. The difficulty during parturition may be associated with the presence of an abnormally enlarged placenta as maternal necropsy findings and microscopic evaluation of the organs did not identify any other factors. In the absence of any other death in the intermediate and high dose groups these deaths are not attributed to the test material.

CLINICAL SIGNS:
Over activity was observed as a post dosing sign during Week 1 in females dosed at 60 mg/kg/day.

BODY WEIGHT AND FOOD CONSUMPTION:
There were no statistically significant effects on bodyweight or bodyweight gain. Weight gain of females from Week 0-5 were slightly lower than Control at all dose levels, but in the absence of any consistent trends it was considered to be unaffected by the test material. During gestation there was no clear effect on bodyweight although gains were slightly lower than Control, and during lactation bodyweight gain of females receiving 250 mg/kg/day were lower than Control.

FOOD CONSUMPTION:
There were no test material related effects on food consumption. The increase in food consumption observed in all animals during the recovery period was due to cessation of dose administration which used corn oil as the vehicle thereby supplying a portion of the required nutrients.

WATER CONSUMPTION:
There were no statistically significant effects observed.

REPRODUCTIVE FUNCTION (ESTROUS CYCLE) AND REPRODUCTIVE PERFORMANCE:
There was no effect of on oestrous cycles or precoital interval. There was no effect on mating performance or fertility at dose levels of 250 mg/kg/day or below. All females that littered had normal length gestation periods (22 – 23 days duration) but a slightly higher proportion of females at 250 mg/kg/day gave birth after 23 days.

ORGAN WEIGHTS:
There were no significant organ weight effects observed.

GROSS PATHOLOGY:
There were no significant necropsy findings for females on Day 7 of lactation.

SIGNS AND ARENA OBSERVATIONS:
There were no signs observed among treated females at routine physical examination or during the arena observations that were considered to be related to treatment.

SENSORY REACTIVITY OBSERVATIONS AND GRIP STRENGTH:
Sensory reactivity observations and grip strength values for Toxicity subgroup animals were similar to those for Controls, and considered unaffected by treatment.

MOTOR ACTIVITY:
Motor activity scores for females showed considerable inter-group variation but no clear dose related trends such that an association with test material was considered unlikely.

HAEMATOLOGY:
There were no marked effects upon haematology parameters.

BLOOD CHEMISTRY:
Glucose plasma levels were significantly higher than Control in females dosed at 250 mg/kg/day. Bile acid plasma levels for females at all dose levels were higher than the concurrent Control. A dose related trend was apparent, however, individual values were all within the Historical Control range (90 percentile range – females: 8.7-49.7 (n=38)).
Dose descriptor:
NOAEL
Effect level:
>= 750 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEL
Effect level:
>= 250 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Details on embryotoxic / teratogenic effects:
LITTER SIZE, SEX RATIO AND SURVIVAL INDICES:
The numbers of implantations, total litter sizes and live litter sizes up to Day 7 of lactation were unaffected by dosing in the 60 and 250 mg/kg/day groups. Sex ratio (assessed by the percentage of males) at 250 mg/kg/day was slightly, but statistically significantly lower than Control. Individual litter data for this parameter are always variable and as only two litters are outside the concurrent Control data range this intergroup difference is not considered to be toxicologically significant. Sex ratio was unaffected at a dose level of 60 mg/kg/day. Administration with CAS 8000-41-7 had no effect on post implantation survival index, live birth index and viability index for animals receiving up to 250 mg/kg/day.

CLINICAL SIGNS: Clinical signs of offspring did not indicate any reaction to maternal exposure by CAS 8000-41-7.

BODY WEIGHT: Male and female offspring bodyweights were considered to be unaffected by CAS 8000-41-7.

GROSS PATHOLOGY: Necropsy findings of offspring killed or dying prior to scheduled termination, and of those killed at the end of the study, did not indicate any reaction to maternal dosing with CAS 8000-41-7.
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
Based on the findings in this study, the NOAEL for maternal and developmental toxicity was >250 mg/kg bw/day.
Executive summary:

In a GLP study conducted according to OECD guideline 422, three groups of ten female rats for the Main (reproductive) phase and five female rats for the Toxicity phase received CAS 8000-41-7 at doses of 0, 60 or 250 mg/kg bw/day in corn oil at a dose volume of 5 mL/kg bw/day. Toxicity phase females were dosed daily for a minimum of five consecutive weeks. Main phase females were dosed daily for two weeks before pairing, throughout mating, gestation and until Day 6 of lactation. During the study, data was recorded on clinical condition, performance under detailed physical and arena examination, sensory reactivity, grip strength, motor activity, bodyweight, food consumption, water consumption (visual), haematology, blood chemistry, oestrous cycles, mating performance and fertility and gestation length. Organ weight, macroscopic and microscopic pathology investigations were undertaken in the adults. The clinical condition of offspring, litter size and survival, sex ratio and offspring bodyweight were assessed and macroscopic pathology investigations were undertaken. In the 60 mg/kg/day dose group, one female was killed because of parturition difficulties. No significant findings were recorded for clinical signs, detailed physical examination and arena observations. Behavioural testing during Week 5 of dosing, including sensory reactivity findings, grip strength values and motor activity scores showed no differences considered to be associated with exposure to the test material. There were no clear effects on bodyweight in unmated females receiving up to 250 mg/kg/day. Bodyweight and bodyweight gain were also unaffected during gestation. During lactation females receiving 250 mg/kg/day showed lower weight gain than Controls. There were no adverse effects on food consumption and water consumption. Also no effects were observed on organ weight and gross pathology. There were no clinically significant effects on haematology parameters, but females showed slight anaemia. Glucose plasma levels were significantly higher than Control in females dosed at 250 mg/kg/day. There were no effects observed on oestrous cycles, precoital interval or mating. Gestation length was within the normal range but there was a small increase in the numbers of animals at 250 mg/kg/day having longer (23 day) gestation periods. At all dose levels there were no effects of the test material on the number of implantations, post implantation survival index, live birth index, viability index and lactation index. Male and female offspring bodyweights, clinical signs, and gross pathology were not adversely affected. Based on the findings in this study, the No-Observed-Adverse–Effect-Level (NOAEL) for maternal and developmental toxicity was at least 250 mg/kg/day.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Study duration:
subacute
Species:
rat
Quality of whole database:
The information from the OECD TG 422 is added to the information included in the developmental toxicity waiver. Together this information is sufficiently adequate to cover this endpoint.
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

For alpha-Terpinyl Acetate no developmental toxicity data are available. There is a combined repeated dose toxicity study with reproduction / developmental toxicity screening available (OECD TG 422) for Terpineol multi, which is summarised in the fertility section. This data can be used for read across (see adequate documentation in the read across section below). In addition, in the waiver for developmental toxicity it is presented that other developmental toxicity than in the OECD TG 422 is not expected.

Alpha-Terpinyl Acetate’s reproductive/developmental toxicity using read across from Terpineol multi

1 Introduction and hypothesis for the analogue approach

The target substance alpha-Terpinyl acetate will be registered in two forms, alpha-Terpinyl Acetate and Terpinyl Acetate multi. For assessing the fertility the repeated dose toxicity information from alpha-Terpinyl Acetate will be used. For other reproductive parameters and developmental toxicity the information from Terpineol multi will be used. The constituents of these substances and their structural similarities are presented in the toxico-kinetic section and will be discussed where needed below. All constituents are structural isomers of each other. For this substance fertility information from a 20-weeks repeated dose toxicity study is available but no information on developmental toxicity. Therefore an analogue approach is applied to fill this gap.

In accordance with Article 13 of REACH, lacking information should be generated whenever possible by means other than vertebrate animal tests, i.e. applying alternative methods such as in vitro tests, QSARs, grouping and read-across. Alpha-Terpinyl Acetate will quickly metabolise into alpha-Terpineol (and this is the main constituent of Terpineol multi).

Hypothesis: alpha-Terpinyl Acetate has developmental toxicity compared to Terpineol multi resulting in similar NOAELs for fertility and developmental toxicity. This is because Terpineol multi has a similar backbone compared to alpha-Terpinyl Acetate and the latter acetic ester will metabolise into alpha-Terpineol when passing the gut and the liver, resulting in a similar reproductive and developmental toxic potential.

Available information: For alpha-Terpinyl Acetate, a reliable 20-weeks dietary repeated dose study is available but predating guidelines and GLP, which will be used for the fertility assessment. For the structurally similar Terpineol multi a combined repeated dose reproductive/developmental toxicity screening test is available, according to OECD TG 422 and under GLP. This test was followed up with 14-day dietary repeated dose studies to judge the effects seen in the OECD TG 422. The OECD TG 422 and the dietary testing will be included for assessing the fertility assessment. The OECD TG 422 conducted with Terpineol multi will be used for read across for screening of the developmental toxicity potential for alpha-Terpinyl Acetate.

2. Target chemical and source chemical(s)

Chemical structures of the target chemical and the source chemicals are shown in the data matrix (see also the attached document in IUCLID section 7.8)

3. Purity / Impurities

The constituents of alpha-Terpinyl Acetate and Terpineol multi are presented in the toxico-kinetic section. The alpha-Terpinyl Acetate is a mono-constituent with a purity of > 80%. This alpha-ester contains gamma- and beta-Terpinyl Acetate, but less than 10% and 1%, respectively. Terpineol multi contains the respective alcohols of these constituents.

4. Analogue approach justification

According to Annex XI 1.5 read across can be used to replace testing when the similarity can be based on a common backbone, a common functional group and/or a common metabolite. When using read across the result derived should be applicable for C&L and/or risk assessment and it should be presented with adequate and reliable documentation.

This read across presented here will be based on common: 1) constituents; 2) structural terpinoid backbones and; 3) alcohol-glucuronated metabolites during systemic exposure.

Structural similarities and differences: The target and the source chemicals have an unsaturated cyclohexyl ring, with a methyl group attached to the unsaturated bond. At the opposite side (C4-position) two methyl group and a tertiary ester (the target and alpha-Terpinyl Acetate) and an alcohol (the source) is attached. The gamma- and beta-constituents of both the ester and the alcohol have a double bond while an alcohol is attached for the alpha-constituent. The alcohol of the gamma- and beta-constituent is attached at the opposite C4-position and has therefore no double bond at the C1-position compared to the alpha-constituent.As has been discussed in the purity section, there are very limited differences between alpha-Terpinyl Acetate and Terpineol multi, considering the constituents. The constituents are all structural isomers of each other. Therefore these minor differences are not expected to influence the toxicity potential.

Toxico-kinetic:Absorption: The source chemicals and the target chemicals are well absorbed via oral, skin and inhalation route based on the similarity in chemical structures and physico-chemical properties. All are liquids. Their vapour pressures are between 3 and 7 Pa. The water solubility of the alcohol is higher and the log Kow lower compared to the ester but still favourable for absorption. Carboxylesterases will metabolise alpha-Terpinyl Acetate completely in the gut and liver into alpha-Terpineol and acetic acid. The alcohols, whether derived from the ester or from the alcohol as such, will all follow the same systemic route (Belsito et al., 2008, WHO, 2000, Wu et al., 2010, and Yamada et al., 2013). This metabolisation of alpha-Terpinyl Acetate into its alcohol and its glucuronation is presented in the toxico-kinetic section. A similar metabolisation pathway is presented in Wu et al (2010) for Terpinyl Propionate who use the metabolisation of this ester as a justified read across between Terpinyl Propionate and alpha-Terpineol. The reproductive/developmental toxicity of alpha-Terpinyl Acetate will therefore result from alpha-Terpineol and the acetic acid. The toxicity of the metabolite acetic acid (Cas no. 64-19-7) did not raise concern for reproductive toxicity (see REACH dissemination tool) as can be expected since acetic acid is a natural component in the body, which will be consumed in the Krebs cycle.

Experimental data on fertility: For fertility the information of the dietary 20-weeks study with alpha-Terpinyl Acetate will be used. In this study no effects were seen on the gonads, indicating absence of effects on fertility at 400 mg/kg bw for both males and females. Supporting information can be derived from Terpineol multi, which contains mainly alpha-Terpineol and which is the metabolite of alpha-Terpinyl Acetate. Terpineol multi has been tested in a combined gavage-dosed repeated dose reproduction / developmental toxicity screening study (OECD TG 422 under GLP, exposure 40-50 days). The target organ in the males was the testis in this OECD TG 422, which showed atrophy and effects on spermatids at 750 mg/kg bw. At the one lower dose, 250 mg/kg bw, no such effects were seen and a NOAEL of 250 mg/kg bw was established for males in this study. For females no fertility effects were seen but the males could not fertilise the females at the dose of 750 mg/kg bw. Therefore the NOAEL for females for fertility was set at >= 750 mg/kg bw. Additional fertility testing on males was performed to investigate the effect of dietary dosing compared to gavage dosing.

When Terpineol multi was dosed via the diet as high as 750 mg/kg bw no testicular effects were seen, which results in the conclusion that the type of dosing (gavage versus dietary) resulted in this effect. This result can be explained by the combined high oral absorption and high dosing. Gavage-dosing result in high peak exposure and can thus results in overloading, the oxygen requiring, glucuronic pathway in the liver, especially when combined with high absorption potential of substances such as the Terpineols. The parent substance (and/or the toxic metabolite) can then enter the systemic circulation instead of being readily excreted via the urine. All organs have the potential to glucuronate substances including the testis. The testis is, however, vulnerable for depletion of oxygen e.g. during glucuronation, because the blood flow in the testicles is low compared to other organs (Aitken and Roman, 2008). This type of testicular effects, including effects on spermatids is related to oxygen depletion by a variety of substances (Aitkin and Roman, 2008). This hypothesis is confirmed with high dietary dosing similar to gavage with Terpineol multi in which no testicular effects were seen. It is also supported by the repeated dose toxicity data of alpha-Terpinyl Acetate in which the rats were dosed much longer (20-weeks) and also no such testicular effects were seen up to a dose of 400 mg/kg bw. For alpha-Terpinyl Acetate the final NOAEL for fertility for both males and females will be >=400 mg/kg bw, because the study period is longer than in the OECD TG 422 and a more appropriate dietary dosing has been used.

Experimental data on developmental toxicity:For developmental toxicity of alpha-Terpinyl Acetate the screening the information from the combined repeated dose reproductive /developmental toxicity of Terpineol multi will be used (OECD TG 422). As has been explained in the toxico-kinetic section and in this read across document, alpha-Terpinyl Acetate will be fully metabolised in the gut and in the liver during normal regular dosing. Therefore the information from its respective alcohol can be used (WHO, 2000, Wu et al., 2010, Yamahara et al., 2013 and). No developmental toxicity effects were seen for Terpineol multi at >= 250 mg/kg bw, the highest dose tested, because of the absence of pregnant dams at 750 mg/kg bw due to testicular effect of the males at 750 mg/kg bw.

Similarities in results for toxicological endpoints between the target and the source chemical(s): It can be seen that alpha-Terpinyl Acetate and Terpineol have a low order of acute and repeated dose toxicity, because no adverse effects are seen up the maximum achievable limit dose. The acetates and the alcohols are also negative for genotoxicity and therefore genotoxicity related developmental toxicity is not expected.

Uncertainty of the prediction on fertility:The testicular effect of Terpineol multi can be well explained using hypothesis of overloading the glucuronic pathway. In case of high gavage peak exposures, this metabolic pathway can require too much oxygen and therefore cause oxygen depletion. Especially, the testis is vulnerable for this effect due to the more limited blood flow compared to other organs. This has been confirmed during dietary testing of Terpineol multi and is supported in the 20-wk study of alpha-Terpinyl Acetate in which no testicular effects were seen. The metabolisation of acetic esters into alcohols is one of the better known and described metabolic pathways (WHO, 2000, Wu et al., 2010 and Yamada et al., 2013) and therefore the likelihood that alpha-Terpinyl Acetate is metabolised into its alcohol is without doubts. Note: Additional research on Terpineol multi is on-going (2012-2013) to investigate the need for doing a one-generation study. When this information becomes available an update of the alpha-Terpinyl Acetate dossier will be done.

Uncertainty of the prediction on developmental toxicity: As has been presented in the paragraph above acetic esters will be fully metabolised in their respective alcohol. Therefore for the developmental screening the information from Terpineol multi can be used for alpha-Terpinyl Acetate. The limitations of the OECD TG 422 for assessing the developmental toxic effects are addressed in the waiver for developmental toxicity.

5. Data matrix

The relevant information on physico-chemical properties and toxicological characteristics are presented in the data matrix below.

6. Conclusions per endpoint for C&L, PBT/vPvB and dose descriptor

When using read across the results derived should be applicable for C&L and/or risk assessment, cover an exposure period duration comparable or longer than the corresponding method and be presented with adequate and reliable documentation.

For assessing the fertility of alpha-Terpinyl Acetate the repeated dose toxicity of its main constituent is used. For alpha-Terpinyl Acetate a well conducted dietary repeated dose toxicity study of 20-weeks study is available in which no testicular or other effects on (female) gonads were seen, resulting in a NOAEL of >=400 mg/kg bw (the highest dose tested). The testicular effects of Terpineol multi during a gavage-dosed reproduction/developmental toxicity screening study can be disregarded being a result of gavage dosing because these effects are not seen during dietary testing.

In summary: Alpha-Terpinyl Acetate does not show adverse effects on fertility.

Developmental toxicity: No hazards were seen at >= 250 mg/kg bw for Terpineol multi, containing mainly alpha-Terpineol and being the main metabolite of alpha-Terpinyl Acetate, in an OECD TG 422. Therefore no such effects are expected for alpha-Terpineol Acetate. A NOAEL for these effects has been set to >=250 mg/kg bw (the highest dose tested). Other information on this endpoint is added in the developmental toxicity waiver.

Final conclusion on hazard, C&L, DNEL and risk characterisation for fertility

Alpha-Terpinyl Acetate has a NOAEL for fertility of >=400 mg/kg bw, which is supported by all information available from Terpineol multi. Classification and labelling is not needed for this endpoint according to EU Directive 67/548/EEC and CLP Regulation (EC) No. 1272/2008. This information also shows that it will not result in a PBT substance and for the hazard assessment conclusion the “no hazard identified” will be taken forward to the risk characterisation.

Final conclusion on hazard, C&L, DNEL and risk characterisation on developmental toxicity

For Terpineol multi a NOAEL of >=250 mg/kg bw has been derived (the highest dose tested). Therefore alpha-Terpinyl Acetate does not need to be classified and labelled for this endpoint. This information and the information in the waiver for the OECD TG 414 the final conclusion is that the substance does not need to be classified and labelled according toEU Directive 67/548/EEC and CLP Regulation (EC) No. 1272/2008, it is not a PBT substance and the for the hazard assessment conclusion “no hazard identified’ will be selected which will be taken forward to the risk characterisation.

References

Aitken, R.J., and Roman, S.D., 2008, Antioxidant systems and oxidative stress in the testis, Oxid. Med. Cell., Longev., 1, 15-24 (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2715191/).

Belsito, D., Bickers, D., Bruze, M., Calow, P., Greim, H., Hanifin, J.M., Rogers, A.E., Saurat, J.H., Sipes, I.G., Tagami, H., 2008, A toxicologic and dermatologic assessment of cyclic acetates when used as fragrance ingredients, Food and Chemical Toxicology 46, Suppl 12:S1-27.

Yamada, T., Tanaka, Y., Hasegawa, R., Sakuratani, Y., Yamada, J., Kamata, E., Ono, A., Hirose., A.,

Wu, S., Blackburn, K., Amburgery, J., Jaworska, J., and Federle, T., 2010, A framework for using structural, reactivity, metabolic and physico-chemical similarity to evaluate the suitability of analogs for SAR-based toxicological assessments, Regul. Toxicol. Pharmacol., 56, 67-81.

Yamada, T., Tanaka, Y., Hasegawa, R., Sakuratani, Y., Yamada, J., Kamata, E., Ono, A., Hirose., A., Yamazoe, Y., Mekenyan, O., Hayashi, M., 2013, A category approach to predicting the repeated-dose hepatotoxicity of allyl esters, Reg. Toxicol. Pharmacol, 65, 189-195.

Data matrix on relevant human health information of alpha-Terpinyl Acetate, Terpineol multi and alpha-Terpineol

Common names

Alpha-Terpinyl Acetate*

Terpineol multi*

Alpha-Terpineol*

Chemical structures#

CAS no

80-26-2

8000-41-7

98-55-5

 

Target

Source

Source Supporting

Molecular weight

196

154

154

Appearance

Liquid

Liquid

Liquid

Physico-chemical data

 

 

 

Melting point,oC

-20

-36

-20

Vapour pressure, Pa

3.5

300^

6.5

Water solubility, mg/l

23

2540

2870

Log Kow

4.4

2.6

2.6

Human health endpoints

 

 

 

Acute oral toxicity in mg/kg bw

5075

(Similar to OECD TG 401)

2000

Read acrossfrom Terpineol multi

Genotoxicity – Ames test

Negative

(OECD TG 471)

Negative

(OECD 471)

Negative

(OECD TG 471)

Repeated dose toxicity mg/kg bw

NOAEL: >= 400

(similar to OECD TG 408)

NOAEL >= 750

(OECD TG 422)

 

Read across from alpha-Terpinyl Acetate and Terpineol multi

Reproductive toxicity: Fertility in mg/kg bw

 

Developmental toxicity

NOAEL of >=400 (similar to OECD TG 408)

 

 

Read acrossfrom Terpineol multi and supporting expert judgment (see waiver)

NOAEL >= 750

 (OECD TG 422 and additional dietary studies)

 

NOAEL >= 250 mg/kg bw (highest dose tested)

Read across from alpha-Terpinyl Acetate

 

 

Read acrossfrom Terpineol multi and supporting expert judgment (see waiver)

# Chemical structures are presented in the attached document. * The constituents of Terpinyl Acetates and the Terpineols are presented in the toxico-kinetic section; ^The 300 Pa value is considered to be an overestimation or due to volatile impurities because the vapour pressure of alpha-Terpineol is 6.5 Pa, which is in line with the MPBPVP QSAR result of 2 Pa.


Justification for selection of Effect on developmental toxicity: via oral route:
The OECD TG 422 of Terpineol multi has been used for read across to alpha-Terpinyl Acetate, because of similarity in the terpinoid backbone and similar systemic exposure, because alpha-Terpinyl Acetate will fully metabolise in alpha-Terpineol, which is the main constituent of Terpineol multi. Therefore this information can also be used for screening for developmental toxicity for the acetate.

Justification for classification or non-classification

Fertility: Alpha-Terpineol Acetate has a NOAEL for fertility of >=400 mg/kg bw based on a repeated dose toxicity study which is supported by all information available from Terpineol multi. This is because the effects seen during gavage dosing in the OECD TG 422 are considered secondary effects as mentioned in CLP section 3.7.2.2.1 “Classification is made on the basis of the appropriate criteria, outlined above, and an assessment of the total weight of evidence (see 1.1.1). Classification as a reproductive toxicant is intended to be used for substances which have an intrinsic, specific property to produce an adverse effect on reproduction and substances shall not be so classified if such an effect is produced solely as a non-specific secondary consequence of other toxic effects”. Therefore for alpha-Terpinyl Acetate classification and labelling is not needed for this endpoint according to EU Directive 67/548/EEC for mixtures and CLP Regulation (EC) No. 1272/2008.

Developmental toxicity: The information from Terpineol multi derived from the OECD TG 422 can be used for screening for developmental toxicity. The main constituent of this substance is alpha-Terpineol being the metabolite of alpha-Terpinyl Acetate. This information did not result in classification and labelling for developmental toxicity. In the absence of e. g. structural alerts and absence of genotoxicity no other developmental toxicity is expected. Therefore alpha-Terpinyl Acetate does not have to be classified and labelled for developmental toxicity according to according to EU Directive 67/548/EEC for mixtures and CLP Regulation (EC) No. 1272/2008.

Additional information